BioMed Research International: Infectious Diseases The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test Tue, 15 Apr 2014 14:05:52 +0000 A generic two-step lyssavirus real-time reverse transcriptase polymerase chain reaction (qRT-PCR), based on a nested PCR strategy, was validated for the detection of different lyssavirus species. Primers with 17 to 30% of degenerate bases were used in both consecutive steps. The assay could accurately detect RABV, LBV, MOKV, DUVV, EBLV-1, EBLV-2, and ABLV. In silico sequence alignment showed a functional match with the remaining lyssavirus species. The diagnostic specificity was 100% and the sensitivity proved to be superior to that of the fluorescent antigen test. The limit of detection was ≤1 50% tissue culture infectious dose. The related vesicular stomatitis virus was not recognized, confirming the selectivity for lyssaviruses. The assay was applied to follow the evolution of rabies virus infection in the brain of mice from 0 to 10 days after intranasal inoculation. The obtained RNA curve corresponded well with the curves obtained by a one-step monospecific RABV-qRT-PCR, the fluorescent antigen test, and virus titration. Despite the presence of degenerate bases, the assay proved to be highly sensitive, specific, and reproducible. Vanessa Suin, Florence Nazé, Aurélie Francart, Sophie Lamoral, Stéphane De Craeye, Michael Kalai, and Steven Van Gucht Copyright © 2014 Vanessa Suin et al. All rights reserved. Nonprofessional Phagocytic Cell Receptors Involved in Staphylococcus aureus Internalization Tue, 15 Apr 2014 09:56:18 +0000 Staphylococcus aureus is a successful human and animal pathogen. The majority of infections caused by this pathogen are life threatening, primarily because S. aureus has developed multiple evasion strategies, possesses intracellular persistence for long periods, and targets the skin and soft tissues. Therefore, it is very important to understand the mechanisms employed by S. aureus to colonize and proliferate in these cells. The aim of this review is to describe the recent discoveries concerning the host receptors of nonprofessional phagocytes involved in S. aureus internalization. Most of the knowledge related to the interaction of S. aureus with its host cells has been described in professional phagocytic cells such as macrophages. Here, we showed that in nonprofessional phagocytes the α5β1 integrin host receptor, chaperons, and the scavenger receptor CD36 are the main receptors employed during S. aureus internalization. The characterization and identification of new bacterial effectors and the host cell receptors involved will undoubtedly lead to new discoveries with beneficial purposes. Nayeli Alva-Murillo, Joel Edmundo López-Meza, and Alejandra Ochoa-Zarzosa Copyright © 2014 Nayeli Alva-Murillo et al. All rights reserved. Comparison of a New Gold Immunochromatographic Assay for the Rapid Diagnosis of the Novel Influenza A (H7N9) Virus with Cell Culture and a Real-Time Reverse-Transcription PCR Assay Mon, 14 Apr 2014 17:27:16 +0000 We assessed a colloidal gold immunochromatographic assay (GICA) for rapid detection of influenza A (H7N9) and compared it with reverse-transcription-polymerase chain reaction (RT-PCR) and viral culture. Samples from 35 H7N9 infected patients were collected, including 45 throat swab samples, 56 sputum samples, and 39 feces samples. All samples were tested by GICA, viral culture, and RT-PCR. GICA specifically reacted with recombinant HA proteins, virus lysates, and clinical samples from H7 subtype viruses. Compared with RT-PCR, GICA demonstrated low sensitivity (33.33%) but high specificity (97.56%). The positive rate of GICA tests for samples collected in the period from 8 to 21 days after contact with poultry was much higher than those for samples collected before or after this period. Compared with viral culture, GICA showed sensitivity of 91.67% and specificity of 82.03%. Sputum specimens were more likely to test positive for H7N9 virus than samples from throat swabs and feces. The GICA-based H7 test is a reliable, rapid, and convenient method for the screening and diagnosis of influenza A (H7N9) disease, especially for the sputum specimens with high viral load. It may be helpful in managing H7N9 epidemics and preliminary diagnosis in early stages in resource-limited settings. Changzhong Jin, Nanping Wu, Xiaorong Peng, Hangping Yao, Xiangyun Lu, Yu Chen, Haibo Wu, Tiansheng Xie, Linfang Cheng, Fumin Liu, Keren Kang, Shixing Tang, and Lanjuan Li Copyright © 2014 Changzhong Jin et al. All rights reserved. Development of a Colloidal Gold Kit for the Diagnosis of Severe Fever with Thrombocytopenia Syndrome Virus Infection Mon, 14 Apr 2014 09:08:17 +0000 It is critical to develop a cost-effective detection kit for rapid diagnosis and on-site detection of severe fever with thrombocytopenia syndrome virus (SFTSV) infection. Here, an immunochromatographic assay (ICA) to detect SFTSV infection is described. The ICA uses gold nanoparticles coated with recombinant SFTSV for the simultaneous detection of IgG and IgM antibodies to SFTSV. The ICA was developed and evaluated by using positive sera samples of SFTSV infection collected from the CDC of China. The reference laboratory diagnosis of SFTSV infection was based on the “gold standard”. The results demonstrated that the positive coincidence rate and negative coincidence rate were determined to be 98.4% and 100% for IgM and 96.7% and 98.6% for IgG, respectively. The kit showed good selectivity for detection of SFTSV-specific IgG and IgM with no interference from positive sera samples of Japanese encephalitis virus infection, Dengue virus infection, Hantavirus infection, HIV infection, HBV surface antigen, HCV antibody, Mycobacterium tuberculosis antibody, or RF. Based on these results, the ICS test developed may be a suitable tool for rapid on-site testing for SFTSV infections. Xianguo Wang, Quanfu Zhang, Fen Hao, Xunian Gao, Wei Wu, Minyao Liang, Zhihua Liao, Shuhong Luo, Weiwen Xu, Dexin Li, and Shiwen Wang Copyright © 2014 Xianguo Wang et al. All rights reserved. What Is New in Listeriosis? Mon, 14 Apr 2014 00:00:00 +0000 Listeriosis is a disease caused by Listeria monocytogenes (L. monocytogenes). L. monocytogenes is bacteria that usually infects some determined inhabitants, especially high risk patients such as the elderly, immunosuppressed patients and pregnant women. However, it can also affect people who do not have these risk factors. L. monocytogenes is widespread in nature being part of the faecal flora of many mammals and it is a common foodborne source. It is acquired by humans primarily through consumption of contaminated food. Besides, between 1% and 10% of the population is a faecal carrier of L.monocytogenes. Listeriosis may occur sporadically or in outbreaks. Infection causes a spectrum of illness, ranging from febrile gastroenteritis to invasive disease, including bacteraemia, sepsis, and meningoencephalitis. This infection has a low incidence, although it is undeniably increasing, particularly due to the rise of population of over 60 years old or of under 60 years olds with a predisposing condition. The diagnosis is complicated because of its incubation period and the different clinical manifestations. Also listeriosis has a high mortality despite adequate and early treatment. The importance of bacteraemia for L. monocytogenes lies in the infrequency of this bacterium and the high mortality, even with appropriate antibiotic treatment. Almudena Hernandez-Milian and Antoni Payeras-Cifre Copyright © 2014 Almudena Hernandez-Milian and Antoni Payeras-Cifre. All rights reserved. Human Antibody Response to Aedes albopictus Salivary Proteins: A Potential Biomarker to Evaluate the Efficacy of Vector Control in an Area of Chikungunya and Dengue Virus Transmission Sun, 13 Apr 2014 13:35:17 +0000 Aedes borne viruses represent public health problems in southern countries and threat to emerge in the developed world. Their control is currently based on vector population control. Much effort is being devoted to develop new tools to control such arbovirus. Recent findings suggest that the evaluation of human antibody (Ab) response to arthropod salivary proteins is relevant to measuring the level of human exposure to mosquito bites. Using an immunoepidemiological approach, the present study aimed to assess the usefulness of the salivary biomarker for measuring the efficacy of Ae. albopictus control strategies in La Reunion urban area. The antisaliva Ab response of adult humans exposed to Ae. albopictus was evaluatedbefore and after vector control measures. Our results showed a significant correlation between antisaliva Ab response and the level of exposure to vectors bites. The decrease of Ae. albopictus density has been detected by this biomarker two weeks after the implementation of control measures, suggesting its potential usefulness for evaluating control strategies in a short time period. The identification of species specific salivary proteins/peptides should improve the use of this biomarker. Souleymane Doucoure, François Mouchet, Sylvie Cornelie, Papa Makhtar Drame, Eric D’Ortenzio, Jean Sébastien DeHecq, and Franck Remoue Copyright © 2014 Souleymane Doucoure et al. All rights reserved. Insights on the Role of Antimicrobial Cuffed Endotracheal Tubes in Preventing Transtracheal Transmission of VAP Pathogens from an In Vitro Model of Microaspiration and Microbial Proliferation Thu, 10 Apr 2014 14:08:47 +0000 We developed an in vitro model to evaluate the effect of different cuffed endotracheal tubes (ETTs) on transtracheal transmission of ventilator-associated pneumonia (VAP) pathogens along external surfaces of ETTs. The model independently assessed the relative contributions of microbial proliferation to the distal tip and microaspiration of contaminated secretions past the cuff by testing in three modes: microaspiration only, microbial proliferation only, and simultaneous microaspiration and microbial proliferation. We evaluated transmission of methicillin resistant Staphylococcus aureus (MRSA) or Pseudomonas aeruginosa (PA) in the presence of a standard ETT; a soft, tapered cuff ETT with subglottic suctioning; and a novel antimicrobial gendine (combination of gentian violet and chlorhexidine) ETT in the model. In the microaspiration only mode, when leakage past the cuff occurred quickly, no ETT prevented transmission. When microaspiration was delayed, the gendine ETT was able to completely disinfect the fluid above the cuff and thereby prevent transmission of pathogens. In microbial proliferation only mode, the gendine ETT was the sole ETT that prevented transmission. With both mechanisms simultaneously available, transmission was dependent on how long microaspiration was delayed. Potent antimicrobial ETTs, such as a gendine ETT, can make unique contributions to prevent VAP when microaspiration is gradual. Joel Rosenblatt, Ruth Reitzel, Ying Jiang, Ray Hachem, and Issam Raad Copyright © 2014 Joel Rosenblatt et al. All rights reserved. Interactions of Neuropathogenic Escherichia coli K1 (RS218) and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells Thu, 10 Apr 2014 10:04:34 +0000 Here we determined the role of various genomic islands in E. coli K1 interactions with phagocytic A. castellanii and nonphagocytic brain microvascular endothelial cells. The findings revealed that the genomic islands deletion mutants of RS218 related to toxins (peptide toxin, α-hemolysin), adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin), protein secretion system (T1SS for hemolysin), invasins (IbeA, CNF1), metabolism (D-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism) showed reduced interactions with both A. castellanii and brain microvascular endothelial cells. Interestingly, the deletion of RS218-derived genomic island 21 containing adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin), protein secretion system (T1SS for hemolysin), invasins (CNF1), metabolism (D-serine catabolism) abolished E. coli K1-mediated HBMEC cytotoxicity in a CNF1-independent manner. Therefore, the characterization of these genomic islands should reveal mechanisms of evolutionary gain for E. coli K1 pathogenicity. Farzana Abubakar Yousuf, Zuhair Yousuf, Junaid Iqbal, Ruqaiyyah Siddiqui, Hafsa Khan, and Naveed Ahmed Khan Copyright © 2014 Farzana Abubakar Yousuf et al. All rights reserved. Phylogenetic Analysis of Stenotrophomonas spp. Isolates Contributes to the Identification of Nosocomial and Community-Acquired Infections Thu, 10 Apr 2014 09:08:14 +0000 Stenotrophomonas ssp. has a wide environmental distribution and is also found as an opportunistic pathogen, causing nosocomial or community-acquired infections. One species, S. maltophilia, presents multidrug resistance and has been associated with serious infections in pediatric and immunocompromised patients. Therefore, it is relevant to conduct resistance profile and phylogenetic studies in clinical isolates for identifying infection origins and isolates with augmented pathogenic potential. Here, multilocus sequence typing was performed for phylogenetic analysis of nosocomial isolates of Stenotrophomonas spp. and, environmental and clinical strains of S. maltophilia. Biochemical and multidrug resistance profiles of nosocomial and clinical strains were determined. The inferred phylogenetic profile showed high clonal variability, what correlates with the adaptability process of Stenotrophomonas to different habitats. Two clinical isolates subgroups of S. maltophilia sharing high phylogenetic homogeneity presented intergroup recombination, thus indicating the high permittivity to horizontal gene transfer, a mechanism involved in the acquisition of antibiotic resistance and expression of virulence factors. For most of the clinical strains, phylogenetic inference was made using only partial ppsA gene sequence. Therefore, the sequencing of just one specific fragment of this gene would allow, in many cases, determining whether the infection with S. maltophilia was nosocomial or community-acquired. Vinicius Godoy Cerezer, Silvia Yumi Bando, Jacyr Pasternak, Marcia Regina Franzolin, and Carlos Alberto Moreira-Filho Copyright © 2014 Vinicius Godoy Cerezer et al. All rights reserved. PCR and Microscopic Identification of Isolated Leishmania tropica from Clinical Samples of Cutaneous Leishmaniasis in Human Population of Kohat Region in Khyber Pakhtunkhwa Tue, 01 Apr 2014 16:21:24 +0000 Leishmania tropica was isolated from the clinical patients of cutaneous leishmaniasis in rural community of Kohat district in Khyber Pakhtunkhwa province and was identified through PCR, microscopy, and culture techniques. A total of 113 samples from the clinical patients were examined through PCR, microscopy, and culture which showed 87.61% (99/113), 53.98% (61/113), and 46.90% (53/113) prevalence. During the study, 186 bp Leishmania tropica was identified through PCR. Thus the sensitivity of PCR is very high as compared to the conventional techniques. Nasser M. Abd El-Salam, Sultan Ayaz, and Riaz Ullah Copyright © 2014 Nasser M. Abd El-Salam et al. All rights reserved. The Function of CD3+CD56+ NKT-Like Cells in HIV-Infected Individuals Thu, 20 Mar 2014 08:42:03 +0000 CD3+CD56+ NKT-like cells are one of the critical effectors in the immune response to viral infection and tumors, but the functional features of NKT-like cells in HIV infection have been rarely reported. In this study, we observed and described the state of NKT-like cell functions in primary HIV-infected individuals (PHIs), chronic HIV-infected individuals (CHIs), long-term nonprogressors (LTNPs), and HIV-negative controls (NCs). The results showed that the percentage of IFN-γ+CD3+CD56+ NKT-like cells was notably higher in LTNPs compared with CHIs, and the proportion of CD3+CD56+ NKT-like cells with dual function (IFN-γ+CD107a+ NKT-like cells) in LTNPs was also much higher than in CHIs. Additionally, the percentages of IFN-γ+CD107a+ NKT-like cells negatively correlated with viral load. Taken together, our data demonstrated that good functions of CD3+CD56+ NKT-like cells in LTNPs likely occurred as a protective mechanism that slows down HIV disease progression. Yongjun Jiang, Xiaojian Cui, Chen Cui, Jian Zhang, Fangyuan Zhou, Zining Zhang, Yajing Fu, Junjie Xu, Zhenxing Chu, Jing Liu, Xiaoxu Han, Christina Liao, Yanan Wang, Yaming Cao, and Hong Shang Copyright © 2014 Yongjun Jiang et al. All rights reserved. TB Meningitis in HIV-Positive Patients in Europe and Argentina: Clinical Outcome and Factors Associated with Mortality Tue, 31 Dec 2013 10:18:23 +0000 Objectives. The study aimed at describing characteristics and outcome of tuberculous meningitis (TBM) in HIV-positive patients and comparing these parameters with those of extrapulmonary TB (TBEP) and pulmonary TB (TBP). Methods. Kaplan-Meier estimation and Poisson regression models were used to assess the mortality following TB diagnosis and to evaluate potential prognostic factors for the 3 groups of TB patients separately. Results. A total of 100 patients with TBM, 601 with TBEP, and 371 TBP were included. Patients with TBM had lower CD4 cell counts and only 17.0% received antiretroviral therapy (ART) at TB diagnosis. The cumulative probability of death at 12 months following TB was 51.2% for TBM (95% CI 41.4–61.6%), 12.3% for TBP (8.9–15.7%), and 19.4% for TBEP (16.1–22.6) (; log-rank test). For TBM, factors associated with a poorer prognosis were not being on ART (adjusted incidence rate ratio (aIRR) 4.00 (1.72–9.09), a prior AIDS diagnosis ( (2.61–8.92)), and receiving care in Eastern Europe ( (2.58–11.34))). Conclusions. TBM among HIV-positive patients was associated with a high mortality rate, especially for patients from Eastern Europe and patients with advanced HIV-infection, which urgently calls for public health interventions to improve both TB and HIV aspects of patient management. Anne Marie W. Efsen, Alexander M. Panteleev, Daniel Grint, Daria N. Podlekareva, Anna Vassilenko, Aza Rakhmanova, Indra Zeltina, Marcelo H. Losso, Robert F. Miller, Enrico Girardi, Joan Caylá, Frank A. Post, Jose M. Miro, Mathias Bruyand, Hansjakob Furrer, Niels Obel, Jens D. Lundgren, Amanda Mocroft, Ole Kirk, and HIV/TB Study Group Copyright © 2013 Anne Marie W. Efsen et al. All rights reserved. Pediatric Tuberculosis in Young Children in India: A Prospective Study Tue, 10 Dec 2013 09:28:33 +0000 Background. India has one of the highest tuberculosis (TB) burdens globally. However, few studies have focused on TB in young children, a vulnerable population, where lack of early diagnosis results in poor outcomes. Methods. Young children (≤5 years) with suspected TB were prospectively enrolled at a tertiary hospital in Pune, India. Detailed clinical evaluation, HIV testing, mycobacterial cultures, and drug susceptibility testing were performed. Results. 223 children with suspected TB were enrolled. The median age was 31 months, 46% were female, 86% had received BCG, 57% were malnourished, and 10% were HIV positive. 12% had TB disease (definite or probable), 35% did not have TB, while TB could not be ruled out in 53%. Extrapulmonary disease was noted in 46%, which was predominantly meningeal. Tuberculin skin test (TST) was positive in 20% of children with TB. Four of 7 (57%) children with culture-confirmed TB harbored drug-resistant (DR) strains of whom 2 (50%) were multi-DR (MDR). In adjusted analyses, HIV infection, positive TST, and exposure to household smoke were found to be significantly associated with children with TB (). Mortality (at 1 year) was 3 of 26 (12%) and 1 of 79 (1%), respectively, in children with TB and those without TB (). Conclusions. Diagnosis of TB is challenging in young children, with high rates of extra-pulmonary and meningeal disease. While the data on DR-TB are limited by the small sample size, they are however concerning, and additional studies are needed to more accurately define the prevalence of DR strains in this vulnerable population. Sanjay K. Jain, Alvaro Ordonez, Aarti Kinikar, Nikhil Gupte, Madhuri Thakar, Vidya Mave, Jennifer Jubulis, Sujata Dharmshale, Shailaja Desai, Swarupa Hatolkar, Anju Kagal, Ajit Lalvani, Amita Gupta, and Renu Bharadwaj Copyright © 2013 Sanjay K. Jain et al. All rights reserved. Circulation of HIV-1 CRF02_AG among MSM Population in Central Italy: A Molecular Epidemiology-Based Study Thu, 28 Nov 2013 17:59:02 +0000 Introduction. The evolutionary and demographic history of the circular recombinant form CRF02_AG in a selected retrospective group of HIV-1 infected men who have sex with men (MSM) resident in Central Italy was investigated. Methods. A total of 55 HIV-1 subtype CRF02_AG pol sequences were analyzed using Bayesian methods and a relaxed molecular clock to reconstruct their dated phylogeny and estimate population dynamics. Results. Dated phylogeny indicated that the HIV-1 CRF02_AG strains currently circulating in Central Italy originated in the early 90's. Bayesian phylogenetic analysis revealed the existence of a main HIV-1 CRF02_AG clade, introduced in the area of Rome before 2000 and subsequently differentiated in two different subclades with a different date of introduction (2000 versus 2005). All the sequences within clusters were interspersed, indicating that the MSM analyzed form a close and restricted network where the individuals, also moving within different clinical centers, attend the same places to meet and exchange sex. Conclusions. It was suggested that the HIV-1 CRF02_AG epidemic entered central Italy in the early 1990s, with a similar trend observed in western Europe. Massimo Giuliani, Maria M. Santoro, Alessandra Lo Presti, Eleonora Cella, Paola Scognamiglio, Alessia Lai, Alessandra Latini, Lavinia Fabeni, Caterina Gori, Carmela Pinnetti, Enrico Girardi, Carlo F. Perno, Gianguglielmo Zehender, and Massimo Ciccozzi Copyright © 2013 Massimo Giuliani et al. All rights reserved. Pattern Recognition Receptors and Cytokines in Mycobacterium tuberculosis Infection—The Double-Edged Sword? Tue, 12 Nov 2013 13:40:38 +0000 Tuberculosis, an infectious disease caused by Mycobacterium tuberculosis (Mtb), remains a major cause of human death worldwide. Innate immunity provides host defense against Mtb. Phagocytosis, characterized by recognition of Mtb by macrophages and dendritic cells (DCs), is the first step of the innate immune defense mechanism. The recognition of Mtb is mediated by pattern recognition receptors (PRRs), expressed on innate immune cells, including toll-like receptors (TLRs), complement receptors, nucleotide oligomerization domain like receptors, dendritic cell-specific intercellular adhesion molecule grabbing nonintegrin (DC-SIGN), mannose receptors, CD14 receptors, scavenger receptors, and FC receptors. Interaction of mycobacterial ligands with PRRs leads macrophages and DCs to secrete selected cytokines, which in turn induce interferon-- (IFN-) dominated immunity. IFN and other cytokines like tumor necrosis factor- (TNF) regulate mycobacterial growth, granuloma formation, and initiation of the adaptive immune response to Mtb and finally provide protection to the host. However, Mtb can evade destruction by antimicrobial defense mechanisms of the innate immune system as some components of the system may promote survival of the bacteria in these cells and facilitate pathogenesis. Thus, although innate immunity components generally play a protective role against Mtb, they may also facilitate Mtb survival. The involvement of selected PRRs and cytokines on these seemingly contradictory roles is discussed. Md. Murad Hossain and Mohd-Nor Norazmi Copyright © 2013 Md. Murad Hossain and Mohd-Nor Norazmi. All rights reserved. Predictors of Delayed Antiretroviral Therapy Initiation, Mortality, and Loss to Followup in HIV Infected Patients Eligible for HIV Treatment: Data from an HIV Cohort Study in India Tue, 29 Oct 2013 15:21:34 +0000 Studies from Sub-Saharan Africa have shown that a substantial number of HIV patients eligible for antiretroviral therapy (ART) do not start treatment. However, data from other low- or middle-income countries are scarce. In this study, we describe the outcomes of 4105 HIV patients who became ART eligible from January 2007 to November 2011 in an HIV cohort study in India. After three years of ART eligibility, 78.4% started ART, 9.3% died before ART initiation, and 10.3% were lost to followup. Diagnosis of tuberculosis, being homeless, lower CD4 count, longer duration of pre-ART care, belonging to a disadvantaged community, being widowed, and not living near a town were associated with delayed ART initiation. Diagnosis of tuberculosis, being homeless, lower CD4 count, shorter duration of pre-ART care, belonging to a disadvantaged community, illiteracy, and age >45 years were associated with mortality. Being homeless, being single, not living near a town, having a CD4 count <150 cells/μL, and shorter duration of pre-ART care were associated with loss to followup. These results highlight the need to improve the timely initiation of ART in HIV programmes in India, especially in ART eligible patients with tuberculosis, low CD4 counts, living in rural areas, or having a low socioeconomic status. Gerardo Alvarez-Uria, Raghavakalyan Pakam, Manoranjan Midde, and Praveen Kumar Naik Copyright © 2013 Gerardo Alvarez-Uria et al. All rights reserved. Evaluation of hsp65 Nested PCR-Restriction Analysis (PRA) for Diagnosing Tuberculosis in a High Burden Country Thu, 24 Oct 2013 10:55:18 +0000 Current study evaluated the hsp65 Nested PCR Restriction Fragment Length Polymorphism Analysis (hsp65 Nested PCR-PRA) to detect and identify Mycobacterium tuberculosis complex directly in clinical samples for a rapid and specific diagnosis of tuberculosis (TB). hsp65 Nested PCR-PRA was applied directly to 218 clinical samples obtained from 127 patients suspected of TB or another mycobacterial infection from July 2009 to July 2010. The hsp65 Nested PCR-PRA showed 100% sensitivity and 95.0 and 93.1% specificity in comparison with culture and microscopy (acid fast bacillus smear), respectively. hsp65 Nested PCR-PRA was shown to be a fast and reliable assay for diagnosing TB, which may contribute towards a fast diagnosis that could help the selection of appropriate chemotherapeutic and early epidemiological management of the cases which are of paramount importance in a high TB burden country. Sara Macente, Clarice Queico Fujimura Leite, Adolfo Carlos Barreto Santos, Vera Lúcia Dias Siqueira, Luzia Neri Cosmo Machado, Nadir Rodrigues Marcondes, Mario Hiroyuki Hirata, Rosário Dominguez Crespo Hirata, and Rosilene Fressatti Cardoso Copyright © 2013 Sara Macente et al. All rights reserved. Frequency and Characteristics of Infections Caused by Extended-Spectrum Beta-Lactamase-Producing Organisms in Neonates: A Prospective Cohort Study Tue, 24 Sep 2013 15:25:39 +0000 This prospective cohort study was conducted to determine the frequency of infections caused by extended-spectrum beta-lactamase- (ESBL-) producing organisms, various bacteria producing ESBL, antibiotic susceptibility of these organisms, and the risk factors associated with these infections in a neonatal intensive care unit in a tertiary care hospital in North India. Of the 150 neonates enrolled in the study, 47 culture-positive neonates were included in the study cohort and were divided into two groups: ESBL-positive (8 neonates) and ESBL-negative (39 neonates) cohorts. Various organisms were isolated from 72 culture samples in these 47 neonates. Of these, 10 culture samples grew ESBL-positive organisms and 62 samples grew ESBL-negative organisms. The frequency of ESBL-producing organisms was found to be 5.3%. ESBL infection incidence densities were found to be 3.4 per 1000 patient-days. Klebsiella (60%) was the most common organism producing ESBL followed by Escherichia coli (30%) and Pseudomonas (10%). Eighty percent of the ESBL-producing organisms were sensitive to piperacillin-tazobactam. Risk factors found significant by univariate analysis () were preterm, low birthweight, perinatal asphyxia, respiratory distress syndrome, anaemia, metabolic acidosis, prolonged mechanical ventilation (>7 days), length of hospitalization, length of level 3 stay, prior antibiotic use, central venous catheter duration, peripherally inserted central venous catheter duration, and total parenteral nutrition duration. Factors that retained significance in the logistic regression model were duration of hospital stay (adjusted OR: 0.958, CI: 0.920–0.997, and value = 0.037) and gestational age (adjusted OR: 1.39, CI: 1.037–1.865, and value = 0.028). There was no significant difference in the mortality between the two groups. Nandini Vijayakanthi, Dheeraj Bahl, Nirmaljit Kaur, Arti Maria, and Nand Kishore Dubey Copyright © 2013 Nandini Vijayakanthi et al. All rights reserved. Hyaluronic Acid Derived from Other Streptococci Supports Streptococcus pneumoniae In Vitro Biofilm Formation Thu, 19 Sep 2013 08:24:42 +0000 We investigate the role of hyaluronic acid (HA) on S. pneumoniae in vitro biofilm formation and evaluate gene expressions of virulence and/or biofilm related genes. Biofilms were grown in medium supplied with HA derived from capsule of Streptococcus equi. The biomasses of biofilms were detected by crystal-violet (CV) microtiter plate assay, and the morphology was viewed under scanning electron microscope (SEM). The gene expressions were assessed by relative quantitative RT-PCR. The results showed that the HA support pneumococcal growth in planktonic form and within biofilms. The CV-microtiter plate assay detected significantly increased biofilm growth in medium containing HA. The SEM analysis revealed thick and organized biofilms in positive control and HA supplemented medium. The nanA, nanB, bgaA, strH, luxS, hysA, ugl, and PST-EIIA encoding genes were significantly upregulated in the planktonic cells grown in presence of HA, while the lytA and comA genes were downregulated. Similarly the luxS, hysA, ugl, and PST-EIIA encoding genes were significantly upregulated by more than 2-folds in HA biofilms. The results of this study indicate that the HA derived from capsule of S. equi supports pneumococcal growth in planktonic state and within biofilms and upregulated virulence and biofilm related genes. Mukesh Kumar Yadav, Sung-Won Chae, Kyeongsoon Park, and Jae-Jun Song Copyright © 2013 Mukesh Kumar Yadav et al. All rights reserved. Listeria monocytogenes Meningitis in Adults: The Czech Republic Experience Wed, 11 Sep 2013 14:05:19 +0000 Background. Listeria monocytogenes (LM) is currently the third most frequent pathogen of bacterial meningitis in adults. Methods. A prospective study of patients with LM meningitis in a Czech tertiary care hospital, carried out from 1997 to 2012. Results. Thirty-one patients were diagnosed with LM meningitis, which was 7% of a total of 440 adult patients with acute bacterial meningitis (ABM) over a 16-year period. Their median age was 63 years, range 26–80 years. Nineteen patients (61%) had underlying immunocompromising comorbidity; 15 patients (48%) were older than 65 years. Fourteen patients (45%) had arterial hypertension. The typical triad of fever, neck stiffness, and altered mental status was present in 21 patients (68%). The median count of cerebrospinal fluid (CSF) leukocytes was 680/L, protein level 2.6 g/L, and glucose ratio 0.28. Four patients (13%) died, and nine (29%) survived with moderate to severe sequelae. Conclusion. LM meningitis is known to affect immunosuppressed and elderly patients. Arterial hypertension seems to be another important predisposing factor. Clinical symptoms, CSF findings, and disease outcomes, did not significantly differ from other community-acquired ABM in our study, although the CSF leukocyte count was lower. Ampicillin showed good clinical and bacteriological efficacy in the majority of patients. Olga Dzupova, Hanus Rozsypal, Dita Smiskova, and Jiri Benes Copyright © 2013 Olga Dzupova et al. All rights reserved. Emergence in Western African Countries of MDR-TB, Focus on Côte d’Ivoire Wed, 04 Sep 2013 13:43:45 +0000 Tuberculosis (TB) is responsible for a high mortality rate (2.5%) worldwide, mainly in developing countries with a high prevalence of human immunodeficiency virus (HIV). The emergence of multiresistant strains of TB poses an extreme risk for TB outbreaks and highlights the need for global TB control strategies. Among Western African countries, Côte d’Ivoire (CI) represents a specific example of a country with great potential to prevent TB. Specifically, CI has a promising healthcare system for monitoring diseases, including vaccination programs. However, military and political conflict in CI favors the spread of infectious diseases, TB being among the most devastating. Compilation of the studies identifying common causes of TB would be extremely beneficial for the development of treatment and prevention strategies. Therefore, the purpose of this comprehensive review is to evaluate the epidemiology of TB in CI, describe the factors involved in pathogenesis, and suggest simple and applicable prevention strategies. Euloge Ekaza, Raymond Kouassi N’Guessan, Adèle Kacou-N’Douba, N’Guetta Aka, Jacquemin Kouakou, Françoise Le Vacon, Fréderic Altare, Gilles Potel, and Marie-France de-La-Cochetiere Copyright © 2013 Euloge Ekaza et al. All rights reserved. Predictors of Attrition in Patients Ineligible for Antiretroviral Therapy after Being Diagnosed with HIV: Data from an HIV Cohort Study in India Thu, 29 Aug 2013 14:50:25 +0000 In newly HIV-diagnosed patients, the CD4+ lymphocyte count is measured to determine the need for antiretroviral therapy (ART). Studies from Sub-Saharan Africa have shown that patients who are ART ineligible at the first assessment have poor retention in care, but data from other low- or middle-income countries are scarce. In this study we describe the retention in pre-ART care of 1696 patients who were ineligible for ART after being diagnosed with HIV in a cohort study in India. More than one-third of ART ineligible patients had poor retention in care, and the attrition was higher in those with longer follow-up periods. Of those patients with poor retention, only 10% came back to the clinics, and their CD4 cell counts were lower than the ones of patients retained in care. After 4.5 years of follow-up, the cumulative incidence of loss to follow-up was 50%. Factors associated with attrition were being homeless, being illiterate, belonging to a disadvantaged community, being symptomatic at the time of the HIV diagnosis, male gender, and not living near a town. Widows were given nutritional support and, therefore, had better retention in care. The results of this study highlight the need to improve the retention in care of ART ineligible patients in India. Gerardo Alvarez-Uria, Manoranjan Midde, Raghavakalyan Pakam, and Praveen Kumar Naik Copyright © 2013 Gerardo Alvarez-Uria et al. All rights reserved. A Qualitative Study of Barriers to Enrollment into Free HIV Care: Perspectives of Never-in-Care HIV-Positive Patients and Providers in Rakai, Uganda Thu, 22 Aug 2013 10:26:08 +0000 Background. Early entry into HIV care is low in Sub-Saharan Africa. In Rakai, about a third (31.5%) of HIV-positive clients who knew their serostatus did not enroll into free care services. This qualitative study explored barriers to entry into care from HIV-positive clients who had never enrolled in care and HIV care providers. Methods. We conducted 48 in-depth interviews among HIV-infected individuals aged 15–49 years, who had not entered care within six months of result receipt and referral for free care. Key-informant interviews were conducted with 12 providers. Interviews were audio-recorded and transcripts subjected to thematic content analysis based on the health belief model. Results. Barriers to using HIV care included fear of stigma and HIV disclosure, women’s lack of support from male partners, demanding work schedules, and high transport costs. Programmatic barriers included fear of antiretroviral drug side effects, long waiting and travel times, and inadequate staff respect for patients. Denial of HIV status, belief in spiritual healing, and absence of AIDS symptoms were also barriers. Conclusion. Targeted interventions to combat stigma, strengthen couple counseling and health education programs, address gender inequalities, and implement patient-friendly and flexible clinic service hours are needed to address barriers to HIV care. Gertrude Nakigozi, Lynn Atuyambe, Moses Kamya, Fredrick E. Makumbi, Larry W. Chang, Neema Nakyanjo, Godfrey Kigozi, Fred Nalugoda, Valerian Kiggundu, David Serwadda, Maria Wawer, and Ronald Gray Copyright © 2013 Gertrude Nakigozi et al. All rights reserved. Mediterranean Spotted Fever in Southeastern Romania Sun, 18 Aug 2013 08:52:32 +0000 Although cases of Mediterranean spotted fever (MSF) have been reported for decades in southeastern Romania, there are few published data. We retrospectively studied 339 patients, diagnosed with MSF at the National Institute of Infectious Diseases “Prof. Dr. Matei Bals” between 2000 and 2011, in order to raise awareness about MSF in certain regions of Romania. According to the Raoult diagnostic criteria 171 (50.4%) had a score >25 points. Mean age was 52.5 years. One hundred and fifty-five (90.6%) patients were from Bucharest and the surrounding region. Almost all patients presented with fever (99.4%) and rash (98.2%), and 57.9% had evidence of a tick bite. There were no recorded deaths. Serologic diagnosis was made by indirect immunofluorescence assay. Of the 171 patients, serology results for R. conorii were available in 147. One hundred and twenty-three (83.7%) of them had a titer IgG ≥1 : 160 or a fourfold increase in titer in paired samples. MSF is endemic in southeastern Romania and should be considered in patients with fever and rash even in the absence of recognized tick exposure. Since the disease is prevalent in areas highly frequented by tourists, travel-associated MSF should be suspected in patients with characteristic symptoms returning from the endemic area. Daniela Pitigoi, Ioana D. Olaru, Daniela Badescu, Alexandru Rafila, Victoria Arama, and Adriana Hristea Copyright © 2013 Daniela Pitigoi et al. All rights reserved. HBV Induced HCC: Major Risk Factors from Genetic to Molecular Level Wed, 07 Aug 2013 15:54:08 +0000 Hepatocellular carcinoma (HCC) is a deadly and emerging disease leading to death in Asian countries. High hepatitis B virus (HBV) load and chronic hepatitis B (CHB) infection increase the risk of developing HCC. HBV is a DNA virus that can integrate DNA into host genome thereby increase the yield of transactivator protein HBxAg that may deregulate many pathways involving in metabolism of cells. Several monogenic and polygenic risk factors are also involved in HCC development. This review summarizes the mechanism involved in HCC development and discusses some promising therapies to make HCC curative. Ambreen Ayub, Usman Ali Ashfaq, and Asma Haque Copyright © 2013 Ambreen Ayub et al. All rights reserved. A Two-Tube Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Sixteen Human Respiratory Virus Types/Subtypes Mon, 05 Aug 2013 09:44:42 +0000 There is a need for the development of a rapid and sensitive diagnosis of respiratory viral pathogens. With an intended application in provincial Centers for Diseases Control and Prevention, in this study, we present a two-tube multiplex RT-PCR assay (two-tube assay) using automatic electrophoresis to simultaneously detect sixteen common respiratory viruses. The specificity and the sensitivity of the assay were tested. The assay could detect 20–200 copies per reaction when each viral type was assayed individually, 2000 copies with 9 premixed viral targets in the multiplexed assay in tube 1, and 200 copies with 8 premixed templates in tube 2. A total of 247 specimens were used to evaluate the two-tube assay, and the results were compared with those obtained from the Luminex xTAG RVP Fast assay. The discordant results were confirmed by sequencing or by the Seeplex RV15 ACE detection kit. There were no false positives, but six false negatives occurred with the two-tube assay. In conclusion, the two-tube assay is demonstrated to have great potential for routine surveillance of respiratory virus infection in China. Jin Li, Shunxiang Qi, Chen Zhang, Xiumei Hu, Hongwei Shen, Mengjie Yang, Ji Wang, Miao Wang, Wenbo Xu, and Xuejun Ma Copyright © 2013 Jin Li et al. All rights reserved. Secondary Bacterial Infections in Patients with Seasonal Influenza A and Pandemic H1N1 Thu, 20 Jun 2013 13:17:01 +0000 The aim of the present study is to analyse the secondary bacterial infections in a large group of patients with seasonal influenza A and influenza A(H1N1) pdm09. Patients diagnosed with seasonal influenza A and influenza A(H1N1) pdm09 between 2005 and 2009 were enrolled in the study. Data was retrieved from medical records and laboratory information systems (LIS). In total, 1094 patients with laboratory confirmed influenza were studied. There were 352 patients with seasonal influenza A and 742 patients with influenza A(H1N1) pdm09. The patients with influenza A were older and had higher comorbidity than patients with influenza A(H1N1) pdm09 ( and , resp.). Hospital admission was higher in influenza A group (). In contrast, ICU admission was higher in patients with influenza A(H1N1) pdm09 than influenza A patients (). There were higher numbers of bacterial samples taken and culture positivity in patients with influenza A than patients with influenza A(H1N1) pdm09 ( and , resp.). In both groups, the majority of the patients with positive bacterial cultures had underlying diseases. The present study shows that the patient characteristics and the frequency of secondary bacterial infections were different in patients with seasonal influenza A and in patients with influenza A(H1N1) pdm09. Karin Liderot, Marcus Ahl, and Volkan Özenci Copyright © 2013 Karin Liderot et al. All rights reserved. Mucosal Leishmaniasis: An Underestimated Presentation of a Neglected Disease Tue, 18 Jun 2013 16:36:22 +0000 We present a review of current knowledge about mucosal leishmaniasis (ML). Although involvement of mucous membranes is classically admitted in New World leishmaniasis, particularly occurring in infection by Leishmania (L.) braziliensis species complex, ML is also a possible presentation of Old World leishmaniasis, in either L. donovani or L. major species complex infections. Thus, ML has to be considered not only as a Latin American disease but as an Old and New World disease. We describe ML epidemiology, pathogenesis, clinics, diagnosis, and therapy. Considering both its highly disfiguring lesions and its possible lethal outcome, ML should not be underestimated by physicians. Moreover, leishmaniasis is expected to increase its burden in many countries as sandfly vector distribution is widespreading towards non-endemic areas. Finally, the lack of clear understanding of ML pathogenesis and the absence of effective human vaccines strongly claim for more research. Alessio Strazzulla, Salvatore Cocuzza, Marilia Rita Pinzone, Maria Concetta Postorino, Stefano Cosentino, Agostino Serra, Bruno Cacopardo, and Giuseppe Nunnari Copyright © 2013 Alessio Strazzulla et al. All rights reserved. NRAMP1, VDR, HLA-DRB1, and HLA-DQB1 Gene Polymorphisms in Susceptibility to Tuberculosis among the Chinese Kazakh Population: A Case-Control Study Mon, 10 Jun 2013 10:01:54 +0000 Background. To explore the potential role of natural-resistance-associated macrophage protein 1 (NRAMP1) gene, vitamin D receptor (VDR) gene, (human leukocyte antigen, (HLA-DRB1) HLA) -DRB1 gene, and HLA-DQB1 gene polymorphisms in susceptibility to tuberculosis (TB) in the Chinese Kazakh population. Methods. A case-control study was performed on the Chinese Kazak population. Genetic polymorphisms of NRAMP1 gene (3′UTR) and VDR gene (TaqI and FokI) were analysed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing analysis in TB patients and healthy controls. Genetic polymorphisms of HLA-DRB1 gene and HLA-DQB1 gene in the two groups were detected with polymerase chain reaction-sequence-specific primers (PCR-SSPs) technique and sequencing analysis. Results. There was statistically significant difference in the 3′UTR polymorphism between the TB patients and healthy controls in the Chinese Kazak population (; OR = 1.859; 95% CI = 1.182–2.926). Significant difference was observed in the FokI polymorphism between the TB patients and healthy controls (; OR = 1.530; 95% CI = 1.007–2.325). It does not disclose any significant association between the disease and TaqI (). Alleles HLA-DRB1*04 and HLA-DQB1*0201 occurred more frequently in patients than in controls ( and 0.002; OR = 1.889 and 1.802; 95% CI = 1.153–3.095 and 1.230–2.639, resp.). Conclusions. Polymorphisms in the NRAMP1 gene, VDR gene, HLA-DRB1 gene, and HLA-DQB1 gene are statistically associated with susceptibility to TB in the Chinese Kazakh population. Fang Wu, Wanjiang Zhang, Le Zhang, Jiangdong Wu, Chunzhu Li, Xianjie Meng, Xi Wang, Peng He, and Jie Zhang Copyright © 2013 Fang Wu et al. All rights reserved. Prevalence and Risk Factors of Toxoplasma gondii in Fattening Pigs Farm from Yucatan, Mexico Thu, 06 Jun 2013 14:30:51 +0000 The objective of this study was to estimate the prevalence and identify risk factors associated with the presence of Toxoplasma gondii in pig-fattening farms from Yucatan, Mexico. A cross-sectional study was conducted with a two-stage sampling. There were 429 pigs sampled from 39 farms randomly selected. Blood samples were collected to obtain DNA and serum. The presence of IgM and IgG antibodies was determined by indirect ELISA. Prevalence was estimated by diagnostic test. Potential risk factors to be included in a marginal logistic regression were tested by chi-square or Fisher. The prevalence of IgM and IgG was 92.5% (397/429) (CI 89.9–95.1%) and 95.8% (411/429) (CI 93.7–97.8%), respectively. Regarding PCR, a prevalence of 50.8% (218/429) (CI 45.9–55.6%) was found. The logistic regression showed an association with herd size and type of feeder (). The risk of a case in farms with ≤400 pigs was 27.9 times higher than in farms with >400 pigs. The manual feeder was a significant protective factor associated with the seropositive against T. gondii. Results indicate a high circulation of T. gondii in pig-fattening farms from Yucatan, finding an increased risk of infection for those farms with less than 400 animals and automatic feeders. A. Ortega-Pacheco, K. Y. Acosta Viana, E. Guzmán-Marín, J. C. Segura-Correa, M. Álvarez-Fleites, and M. Jiménez-Coello Copyright © 2013 A. Ortega-Pacheco et al. All rights reserved. A History of the Development of Brucella Vaccines Mon, 03 Jun 2013 13:46:48 +0000 Brucellosis is a worldwide zoonosis affecting animal and human health. In the last several decades, much research has been performed to develop safer Brucella vaccines to control the disease mainly in animals. Till now, no effective human vaccine is available. The aim of this paper is to review and discuss the importance of methodologies used to develop Brucella vaccines in pursuing this challenge. Eric Daniel Avila-Calderón, Ahidé Lopez-Merino, Nammalwar Sriranganathan, Stephen M. Boyle, and Araceli Contreras-Rodríguez Copyright © 2013 Eric Daniel Avila-Calderón et al. All rights reserved. Mixed Production of Filamentous Fungal Spores for Preventing Soil-Transmitted Helminth Zoonoses: A Preliminary Analysis Sun, 21 Apr 2013 16:20:33 +0000 Helminth zoonoses are parasitic infections shared by humans and animals, being the soil-transmitted helminths (STHs) mainly caused by roundworms (ascarids) and hookworms. This study was aimed to assess the individual and/or mixed production of two helminth-antagonistic fungi, one ovicide (Mucor circinelloides) and other predator (Duddingtonia flagrans). Fungi were grown both in Petri plates and in a submerged culture (composed by water, NaCl, Na2HPO4 · 12 H2O, and wheat (Triticum aestivum)). A Fasciola hepatica recombinant protein (FhrAPS) was incorporated to the cultures to improve fungal production. All the cultured plates showed fungal growth, without difference in the development of the fungi when grown alone or mixed. High counts of Mucor spores were produced in liquid media cultures, and no significant differences were achieved regarding single or mixed cultures, or the incorporation of the FhrAPS. A significantly higher production of Duddingtonia spores after the incorporation of the FhrAPS was observed. When analyzing the parasiticide efficacy of the fungal mixture, viability of T. canis eggs reduced to 51%, and the numbers of third stage cyathostomin larvae reduced to 4%. It is concluded, the capability of a fungal mixture containing an ovicide (Mucor) and a predator species (Duddingtonia) for growing together in a submerged medium containing the FhrAPS offers a very interesting tool for preventing STHs. M. S. Arias, C. F. Cazapal-Monteiro, J. Suárez, S. Miguélez, I. Francisco, F. L. Arroyo, J. L. Suárez, A. Paz-Silva, R. Sánchez-Andrade, and P. Mendoza de Gives Copyright © 2013 M. S. Arias et al. All rights reserved. MLST Genotypes and Antibiotic Resistance of Campylobacter spp. Isolated from Poultry in Grenada Sun, 24 Feb 2013 09:18:27 +0000 This study determined whether multilocus sequence types (MLST) of Campylobacter from poultry in 2 farms in Grenada, West Indies, differed by farm, antimicrobial resistance and farm antibiotic use. Farm A used fluoroquinolones in the water and Farm B used tetracyclines. The E-test was used to determine resistance of isolates to seven antibiotics. PCR of the IpxA gene confirmed species and MLST was used to characterize 38 isolates. All isolates were either C. jejuni or C. coli. Farm antibiotic use directly correlated with antimicrobial resistance of Campylobacter isolates. Almost 80% of the isolates from Farm A were fluoroquinolone resistant and 17.9% of the isolates from Farm B were fluoroquinolone resistant. All Campylobacter isolates from Farm A were tetracycline sensitive, whereas 35.7% of isolates from Farm B were tetracycline resistant. Six previously recognized sequence types (STs) and 2 novel STs were identified. Previously recognized STs were those overwhelmingly reported from poultry and humans globally. Isolates with the same ST did not always have the same antibiotic resistance profile. There was little ST overlap between the farms suggesting that within-farm transmission of Campylobacter genotypes may dominate. MLST typing was useful for tracking Campylobacter spp. among poultry units and can help elucidate Campylobacter host-species population structure and its relevance to human health. Diana Stone, Margaret Davis, Katherine Baker, Tom Besser, Rohini Roopnarine, and Ravindra Sharma Copyright © 2013 Diana Stone et al. All rights reserved. Asymptomatic Malaria Correlates with Anaemia in Pregnant Women at Ouagadougou, Burkina Faso Sun, 11 Nov 2012 17:08:01 +0000 Sub-Saharan Africa records each year about thirty-two million pregnant women living in areas of high transmission of Plasmodium falciparum causing malaria. The aim of this study was to carve out the prevalence of asymptomatic malaria among pregnant women and to emphasize its influence on haematological markers. The prevalence of Plasmodium falciparum asymptomatic infection among pregnant women was 30% and 24% with rapid detection test (RDT) and microscopy, respectively. The prevalence of P. falciparum asymptomatic malaria was reduced among pregnant women using sulfadoxine-pyrimethamine's intermittent preventive treatment and 61% of them were anaemic. Anaemia was significantly more common in women infected with P. falciparum compared with the uninfected pregnant women. Most of the women had normal levels of homocysteine and low levels of folate, respectively. Therefore, the systematic diagnosis of malaria should be introduced to pregnant women as a part of the antenatal care. Zoenabo Douamba, Cyrille Bisseye, Florencia W. Djigma, Tegwinde R. Compaoré, Valérie Jean Telesphore Bazie, Virginio Pietra, Jean-Baptiste Nikiema, and Jacques Simpore Copyright © 2012 Zoenabo Douamba et al. All rights reserved. Epithelial Cell Coculture Models for Studying Infectious Diseases: Benefits and Limitations Wed, 05 Oct 2011 09:12:50 +0000 Countless in vitro cell culture models based on the use of epithelial cell types of single lineages have been characterized and have provided insight into the mechanisms of infection for various microbial pathogens. Diverse culture models based on disease-relevant mucosal epithelial cell types derived from gastrointestinal, genitourinary, and pulmonary organ systems have delineated many key host-pathogen interactions that underlie viral, parasitic, and bacterial disease pathogenesis. An alternative to single lineage epithelial cell monoculture, which offers more flexibility and can overcome some of the limitations of epithelial cell culture models based on only single cell types, is coculture of epithelial cells with other host cell types. Various coculture models have been described, which incorporate epithelial cell types in culture combination with a wide range of other cell types including neutrophils, eosinophils, monocytes, and lymphocytes. This paper will summarize current models of epithelial cell coculture and will discuss the benefits and limitations of epithelial cell coculture for studying host-pathogen dynamics in infectious diseases. Benjamin L. Duell, Allan W. Cripps, Mark A. Schembri, and Glen C. Ulett Copyright © 2011 Benjamin L. Duell et al. All rights reserved. A New Microarray System to Detect Streptococcus pneumoniae Serotypes Thu, 09 Jun 2011 15:06:52 +0000 Streptococcus pneumoniae, one of the most common gram-positive pathogens to colonize the human upper respiratory tract, is responsible for many severe infections, including meningitis and bacteremia. A 23-valent pneumococcal vaccine is available to protect against the 23 S. pneumoniae serotypes responsible for 90% of reported bacteremic infections. Unfortunately, current S. pneumoniae serotype testing requires a large panel of expensive antisera, assay results may be subjective, and serotype cross-reactions are common. For this study, we designed an oligonucleotide-based DNA microarray to identify glycosyltransferase gene sequences specific to each vaccine-related serotype. Out of 56 isolates representing different serotypes, only one isolate, representing serotype 23A, was not detected correctly as it could not be distinguished from serotype 23F. Our data suggest that the microarray provides a more cost-effective and reliable way of monitoring pneumococcal capsular types. Yuka Tomita, Akira Okamoto, Keiko Yamada, Testuya Yagi, Yoshinori Hasegawa, and Michio Ohta Copyright © 2011 Yuka Tomita et al. All rights reserved.