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Journal of Biomedicine and Biotechnology
Volume 2012 (2012), Article ID 435982, 5 pages
http://dx.doi.org/10.1155/2012/435982
Research Article

Loop-Mediated Isothermal Amplification for Detection of Staphylococcus aureus in Dairy Cow Suffering from Mastitis

1The College of Traditional Chinese Veterinary Medicine, Agricultural University of Hebei, Dingzhou 073000, China
2Hebei Engineering and Technology Research Center of Veterinary Biological Products, Agricultural University of Hebei, Baoding 071000, China

Received 24 September 2012; Revised 7 November 2012; Accepted 7 November 2012

Academic Editor: Michael D. Coleman

Copyright © 2012 Zhang Tie et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

To develop a rapid detection method of Staphylococcus aureus using loop-mediated isothermal amplification (LAMP), four specific primers were designed according to six distinct sequences of the nuc gene. In addition, the specificity and sensitivity of LAMP were verified and compared with those of PCR. Results showed that the LAMP reaction was completed within 45 min at 62.5°C, and ladder bands were appeared in LAMP products analyzed by gel electrophoresis. After adding 1x SYBR Green l, the positive reaction tube showed green color and the negative reaction tube remained orange, indicating that the LAMP has high specificity. The minimal detectable concentration of LAMP was  CFU/mL and that of PCR was  CFU/mL, indicating that the LAMP was 100 times more sensitive than the PCR. The LAMP method for detection of Staphylococcus aureus has many advantages, such as simple operation, high sensitivity, high specificity, and rapid analysis. Therefore, this method is more suitable for the rapid on-site detection of Staphylococcus aureus.