Loop-mediated isothermal amplification assay for the detection of Aeromonas hydrophila
MENG Shuang, WANG Yi, WANG Yan, YE Chang-yun
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center of Disease Control and Prevention, Beijing 102206, China
Abstract:To develop a sensitive and specific loop-mediated isothermal amplification(LAMP) assay for the detection of Aeromonas hydrophila, a set of 6 primers targeted toward the desA gene of the A. hydrophila was designed using PrimerExplorer V4 software(Eiken Chemical Co. Ltd., Tokyo, Japan) based on the conserved sequences. The performance of the assay with reference plasmids, simulated human stools, and clinical specimens were evaluated and compared with quantitative PCR(qPCR). No false-positive results were observed for the 33 non-A. hydrophila strains used to evaluate assay specificity. The sensitivity of the LAMP assay was approximately 20 copies per reaction in reference plasmids and 5×103 CFU per gram in spiked human stool, which were more sensitive than the results of qPCR. The performance of the LAMP assay was evaluated with clinical specimens, which had the better detection ability than qPCR. In conclusion, the LAMP assay developed in this study is a valuable method for rapid, cost-effective, and simple detection of A. hydrophila in basic clinical and field laboratories.
孟双, 王毅, 王艳, 叶长芸. 环介导等温扩增技术在嗜水气单胞菌检测中的应用[J]. 中国人兽共患病学报, 2016, 32(1): 1-6.
MENG Shuang, WANG Yi, WANG Yan, YE Chang-yun. Loop-mediated isothermal amplification assay for the detection of Aeromonas hydrophila. Chinese Journal of Zoonoses, 2016, 32(1): 1-6.
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