多重PCR液相芯片技术在腹泻病原体检测中的应用  

Application of multiplex PCR liquid chip technology in the detection of diarrhea pathogens

作　　者：黎俊宏[1] 姚萍[1] 李琼[1] 王凤鸣[1] 杜强[1] 屠博文[1] 吉俊敏[1] 

机构地区：[1]常州市疾病预防控制中心,江苏常州213022

出　　处：《现代预防医学》2017年第18期3390-3394,共5页Modern Preventive Medicine

基　　金：常州市科技支撑计划(社会发展)(项目号:CE20165042);江苏省基础研究计划(自然科学基金)--青年基金项目(项目号:BK20150250)

摘　　要：目的探讨多重PCR液相芯片技术检测腹泻病原体的可行性,为突发公共卫生事件处置提供实验依据。方法采用x TAGRGastrointestinal Pathogen Panel(x TAG GPP)对模拟腹泻阳性样本和783份腹泻疫情样本进行检测,结果与常规检测法比较。结果模拟腹泻阳性样本均检出15种腹泻病原体。783份腹泻疫情标本中未检出艰难梭状芽胞杆菌(Clostridium difficile,C.difficile)、大肠埃希菌O157(Escherichia coli O157,E.coli O157)、微小隐孢子虫(Cryptosporidium,C.parvum)、痢疾阿米巴(Entamoeba histolytica,E.histolytica)和贾第虫(Giardia,Giar)。病毒和细菌检出率分别为55.04%和38.19%;诺如病毒GI/GII型(Norovirus GI/GII,NV GI/GII)、轮状病毒A型(Rotavirus A,RV A)、空肠弯曲菌(Campylobacter jejuni,C.jejuni)、肠毒素性大肠埃希菌LT/ST型(Enterotoxigenic E.coli LT/ST,ETEC)、志贺氏菌(Shigella,Shig)、产志贺毒素大肠埃希菌stx1/stx2型(Shiga-like Toxin producing E.coli stx 1/stx 2,STEC)、小肠结肠炎耶尔森菌(Yersinia enterocolitica,Y.enterocolitica)检出率差异均有统计学意义。结论多重PCR液相芯片技术具有高通量、高灵敏度、高特异性等优点,能较全面评估腹泻病原体和检测混合感染样本,处理好经费和PCR污染的问题,该方法可用于腹泻疫情的快速应急检测。Objective The aim of this study was to investigate the emergency detection feasibility of the multiple PCR liquid chip technology in diarrhea outbreaks, and provide the laboratory basis for the public health emergency disposal. Methods The simulation of diarrhea positive and 783 diarrhea outbreaks samples were detected by xTAGGas trointestinal Pathogen Panel（x TAGRGPP）. And the results were compared with the conventional detection methods. Results Fifteen pathogens were detected in the simulation of diarrhea positive samples. In 783 diarrhea outbreak specimens, Clostridium difficile（C.difficile）, Escherichia coli O157（E. coli O157）, Cryptosporidium（C. parvum）, Entamoeba histolytica（E. histolytica） and Giardia（Giar） were not detected. The viral and bacterial detection rates were 55.04 % and 38.19 %, respectively. Norovirus GI/GII（NV GI/GII）, Rotavirus A（RV A）, Campylobacter jejuni（C. jejuni）, Enterotoxigenic E. coli LT/ST（ETEC）, Shigella（Shig）, Shiga-like toxin producing E. coli stx 1/stx 2（STEC）, Yersinia enterocolitica（Y. enterocolitica） were statistically significant. There were co-infections diarrhea outbreaks pathogens. Conclusion The multiple PCR liquid chip technology can evaluate diarrhea pathogens relative comprehensively and detect mixed infection samples with advantages of high throughput,high sensitivity and high specificity. It can be used for rapid emergency detection in diarrhea outbreaks.

关 键 词：多重PCR 液相芯片 xTAGGPP 腹泻疫情 

分 类 号：R183.4[医药卫生—流行病学]