Highly sensitive and confirmatory Sample 5 L, total time 135 minutes, antigen coated plate HRP and tetramethyl benzidine (substrate) Highly sensitive and confirmatory Detection of JEV positive and negative results in CSF and serum
Can differentiate infection type, intensity, and presence of JEV strains and detects specific IgM in the cerebrospinal fluid or in the blood Detection of immunoglobulin in human serum to JEV- derived antigens. In vitro diagnostic use Status of viral encephalitis, neuroinflammation
MAC-ELISA
MAC ELISA is used to diagnose secondary flavivirus infection
Used to detect true positive and true negative sensitivity and specificity in JEV affected patients
Single TaqMan assay
Highly sensitive and confirmatory
Diagnoses virus antigens
Immunofluorescent test
Highly sensitive and confirmatory
Works as a valuable alternative to the established methods in detecting anti-JEV antibodies after vaccination in travelers and helps in the diagnosis of acutely infected persons, in vitro labeling of NPC cells
Fluoresecent markers are used Fluorescent dye 7-ADD binds to DNA. The labelled cells are detected by FL-1 channel by FACS and are analyzed by using Cell Quest Pro software to quantify percentage of labelled cells.
The amount of flouresecent antibody bound to each cell can be quantified
Florescence resonance energy transfer (FRET)
Highly sensitive
Detects interaction of antigens in cells
Plaque reduction or JEV-antibody neutralization test (PRNT)
Moderately sensitive and confirmatory On incubation, the antibody forming cells release immunoglobulin which coats the surrounding erythrocytes. Complement causes lysis of coated cells and plaque clear or red cells are counted. Hemolytic efficiency of IgM antibodies is detected
Can detect humeral immune response generated after immunization with JE inactivated vaccine Used for viremia determination Percentage neutralization is calculated from number of plaques obtained
Microcomplement fixation test
Moderately sensitive
Can detect cellular factors and antigens
Virus overlay protein binding assay (VOPBA)
Highly sensitive and confirmatory
Detects JE virus receptor molecules on the cells
YUNEL assay
Highly sensitive
Apoptosis, cell membrane disruption, and morphology
Lumbar puncture test and CSF analysis
Moderately sensitive and confirmatory
Probable and confirmed JE
MRI (magnetic resonance imaging)
Moderately sensitive and confirmatory
Can locate bilateral thalamic lesions with hemorrhage, and any abnormality generated in basal ganglia, putamen, pons, spinal cord, and cerebellum may also show pathological abnormalities
CT scan (computed tomography)
Highly sensitive and confirmatory
Can locate hyperintense lesions in the areas of the thalamus, cerebrum, and cerebellum
EEG (electroencephalogram)
Moderately sensitive and confirmatory
Reveals diffuse and burst suppression
CBC (complete blood count)
Confirms the presence of JEV infection in children and helps in clinical analysis of blood parameters
Detect leukocytosis, leucopenia, anemia and thrombocytopenia, and supportive lymphocytic pleocytosis
Platelet count
Sensitive and supportive for clinical analysis
Can detect effect of fever on blood platlets
Hemagglutination inhibition test (HA)
Moderately sensitive Agglutination is done by using antigen coated particles
Antibody detection to detect rheumatoid factors identification of antibodies to soluble antigens. HA is used to detect JEV in various passages
Compliment fixation test (CF) or crosslinking of antigens
Moderately sensitive
Antibody detection. Surface antigens are detected by using labeled antibodies. Both monovalent and divalent antibodies are used
Immunotyping
Highly sensitive and confirmatory
Differentiates genotypes of JE virus
RPHA, IFA, immunoperoxidase
Moderately sensitive
Antigen detection
Immunoblotting
Highly sensitive
JEV generated infection in NCPs and recognizes decrease in the number of colony forming neurosphere and their self-reveal, HRP, PBS-T
IDD (immunodouble diffusion test)
Moderately sensitive
Immunologic relationship between the antigens related or indicative or unrelated Precipitate forms an opaque line in the cross-reactive region
Cell death assay (annexin-propidium iodide staining test)
Highly sensitive
Can recognize apoptotic cell death in control and JEV infected cells. FITC labeled annexin and propidium iodide are used
Nephrometry
Moderately sensitive
Antigen and antibody dilutions are used to create cloudiness, and greater sensitivity can be generated by using monochromatic light from a laser and by adding PEG to solution to increase the size of aggregation
Neutralization tests
Moderately sensitive
Neutralization antibody titre in sera and in CSF can recognize homologous virus, the challenge virus, and the selected wild-type JE virus
Flow cytometry (FACS)
Highly sensitive
Intracellular signaling of JEV antigen, to detect percentage of anti-JEV-FITC positive cells.
Immunohistochemistry
Highly sensitive
Intracellular localization of NS3 by using anti- JEV antibodies
Precipitin test
Moderately sensitive
Quantitative analysis of antigen and antibody interaction
SRID
Highly sensitive
To know the amount of antigen in unknown samples
Neurovirulence test
Highly sensitive
To detect histopathological recognition of JEV pathogenesis in brain and in associated tissues. Prediction of level and cause of neurovirulence
Anticomplementarity Test
Highly sensitive
Identification of lesion scores
Hemolysin test
Moderately sensitive
Percent of hemolysis in RBCs
DNA microarray
Highly sensitive and confirmatory
Expression of genes and proteins
Site directed mutagenesis
Highly confirmatory
Detects amino acid substitutions in E, NS1, and NS2 proteins, clone-specific substitutions, and heterogeneity substitutions and is used to detect possible mutations in structural and non structural viral proteins
Real-time polymerase chain reaction (RT-PCR)
Highly sensitive
Target sequences can be detected in genes and viral genome. Amplification of immunotype strain, cloning, and expression of NS3 gene of NS3 protein of JEV
RNA studies oligonucleotide primer-based detection of JEV functional sequences in different genes and gene copies
Highly sensitive
Detects molecular pathogenesis at the level of enzymes, genes, factors, and proteins. Synthesis and secretion of JEV-induced proteins
Presence of JEV viral and virus secreted antibodies are detected in cerebrospinal fluid (CSF) and serum samples. For component-based detection of JEV, a wide variety of conventional techniques such as viral neutralization, hemagglutination (HI), and complement fixation and immunoflourescent staining are used. Laboratory diagnosis of JE virus is mostly confirmed by immunological, molecular, and biophysical methods. Most of the laboratory-based tests and clinical diagnostic tests are routinely used to detect presence of JEV virus and its pathogenesis but all such tests are labor-intensive, expensive, and cumbersome.
