State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping, Beijing, People's Republic of China.
PLoS One. 2012;7(10):e41978. doi: 10.1371/journal.pone.0041978. Epub 2012 Oct 15.
Plesiomonas shigelloides is one of the causative agents of human gastroenteritis, with increasing number of reports describing such infections in recent years. In this study, the hugA gene was chosen as the target to design loop-mediated isothermal amplification (LAMP) assays for the rapid, specific, and sensitive detection of P. shigelloides. The performance of the assay with reference plasmids and spiked human stools as samples was evaluated and compared with those of quantitative PCR (qPCR). No false-positive results were observed for the 32 non-P. shigelloides strains used to evaluate assay specificity. The limit of detection for P. shigelloides was approximately 20 copies per reaction in reference plasmids and 5×10(3) CFU per gram in spiked human stool, which were more sensitive than the results of qPCR. When applied in human stool samples spiked with 2 low levels of P. shigelloides, the LAMP assays achieved accurate detection after 6-h enrichment. In conclusion, the LAMP assay developed in this study is a valuable method for rapid, cost-effective, and simple detection of P. shigelloides in basic clinical and field laboratories in the rural areas of China.
希氏气单胞菌是人类肠胃炎的病原体之一,近年来越来越多的报告描述了此类感染。在这项研究中,选择 hugA 基因为靶标,设计了环介导等温扩增(LAMP)检测方法,用于快速、特异性和敏感地检测希氏气单胞菌。评估了该检测方法与定量 PCR(qPCR)在参考质粒和加标人粪便样本中的性能。在用于评估检测方法特异性的 32 株非希氏气单胞菌菌株中,未观察到假阳性结果。在参考质粒中,希氏气单胞菌的检测限约为每个反应 20 个拷贝,在加标人粪便中为 5×10(3) CFU/克,比 qPCR 的结果更敏感。当应用于加标有 2 低水平希氏气单胞菌的人粪便样本时,经过 6 小时富集后,LAMP 检测方法可实现准确检测。总之,本研究中开发的 LAMP 检测方法是在中国农村地区基本临床和现场实验室快速、经济高效且简单检测希氏气单胞菌的一种有价值的方法。