Establishment and Application of Loop-mediated Isothermal Amplification Assay for Detection of Staphylococcus aureus
ZHANG Xiao-Yu, WANG Wen-Wen, MA Chen-Jie, WU Xiao-Ling, DENG Guang-Cun*
School of Life Sciences/Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in the Western China, Ningxia University, Yinchuan 750021, China
Abstract：Bovine mastitis is the most harmful common disease in dairy farms, which seriously affects the healthy development of dairy industry. Bacterial infection is main cause of bovine mastitis. Staphylococcus aureus is one of the most common causative pathogen which causing bovine mastitis. Therefore, the rapid detection method for Staphylococcus aureus is of great significance to realize the early prevention of mastitis in dairy cows. The loop-mediated isothermal amplification (LAMP) technique is a new nucleic acid amplification method that could quickly detect target DNA under isothermal conditions. In this study, a set of 6 specific primers were designed according to the nuc gene (GenBank No. DQ507379.1) of S. aureus, and LAMP technique for S. aureus was established by optimizing the reaction conditions, and then the specificity, sensitivity and clinical detection effect of the method were evaluated. The results showed that the established method could specifically detect S. aureus, the minimum detect limit was 2.7×101 copies/μL, and the sensitivity of LAMP detection was 100 times higher than that of PCR method. The LAMP method which had the advantage of simplicity, rapidity, sensitivity and high specificity, was suitable to rapidly detect S. aureus in dairy farms. This study provides a technical reference for the early diagnosis of dairy cow mastitis caused by other pathogenic bacteria.
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