Liang Hao, Wen Ziyu, Li Ying, Duan Yongxiang, Gu Yixin, Zhang Maojun
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Nanshan Center for Disease Control and Prevention, Shenzhen, China.
Front Microbiol. 2018 Dec 5;9:2995. doi: 10.3389/fmicb.2018.02995. eCollection 2018.
is one of the most common pathogens leading to the bacterial diarrheal illness. In order to set up one effective culture independent assay for the screen of the infection in the diarrheal patients, the quadruple real-time PCR method comparing to the culture based on the enriched filtration method which was recognized as the most effective isolation method was assessed for 190 stool samples from the diarrheal patients collected during the Foodborne Diseases Active Surveillance Network in Beijing. This multiple real-time PCR was designed to identify the genus, , and simultaneously. With the enrichment culture method, 23 (12.1%, 23/190) isolates were obtained (20 and 3 ), however, 31 samples (16.3%, 31/190) were detected positively with the real-time PCR (21 , 8 , and 2 genus only). With the comparison, the real-time-PCR method is more sensitive than the enrichment filtration method (16.3 vs. 12.1%, = 0.021). Among the culture-positive samples, 95.7% (22/23) were detected positively by PCR which indicate the specificity of this method was higher. These two methods were consistent well (Kappa = 0.785, < 0.05). Comparing to the culture methods, the result of the multiple real-time PCR method is sensitive, reliable and rapid. The present study indicated this multiple real-time PCR can be used both for the surveillance network and the preceding screen for bacteria isolation. This is first comparative study between the culture and multiple real-time PCR method for identification in stool specimens from the diarrheal patients.
是导致细菌性腹泻疾病最常见的病原体之一。为建立一种有效的非培养检测方法以筛查腹泻患者中的感染情况，对北京市食源性疾病主动监测网络期间收集的190份腹泻患者粪便样本，评估了与基于富集过滤法（被认为是最有效的分离方法）的培养法相比的四重实时荧光定量PCR方法。这种多重实时荧光定量PCR旨在同时鉴定属、和。采用富集培养法，获得了23株（12.1%，23/190）分离株（20株和3株），然而，实时荧光定量PCR检测到31份样本（16.3%，31/190）呈阳性（21株、8株和2株仅属）。相比之下，实时荧光定量PCR方法比富集过滤法更敏感（16.3%对12.1%，P = 0.021）。在培养阳性样本中，95.7%（22/23）通过PCR检测呈阳性，这表明该方法的特异性更高。这两种方法一致性良好（Kappa = 0.785，P < 0.05）。与培养方法相比，多重实时荧光定量PCR方法的结果敏感、可靠且快速。本研究表明这种多重实时荧光定量PCR可用于监测网络和细菌分离的前期筛查。这是首次对腹泻患者粪便标本中鉴定的培养法和多重实时荧光定量PCR方法进行比较研究。
