霍乱、伤寒、痢疾及其他感染性腹泻仍是影响人类健康的重要疾病。目前血清型鉴定是肠道致病菌临床检测和疾病监测的金标准,主要依赖玻片凝集实验。该经典检测手段存在操作繁琐、耗时长、重现性差、价格贵、易受主观因素干扰等诸多缺陷。基于分子生物学的血清型分子鉴定成为最有前途的替代方案。近年国内外报道了多种血清型分子鉴定方法,但实际应用仍有限，原因在于：通量低，覆盖的血清型种类少；芯片或微球技术操作繁琐且易产物污染，且需昂贵的专用设备。本课题组提出的基于荧光探针熔解曲线的多重检测技术（MeltPleX），可在荧光PCR仪上单管一步检测42-48个靶基因。本项目拟用该技术研发针对4种肠道致病菌（霍乱弧菌、志贺菌、副溶血弧菌、致泻性大肠杆菌）的一套集成核酸提取、扩增和检测的一体式快速血清型分子鉴定系统，鉴定结果4小时内，软件自动判读结果。旨在为基层防控重要肠道传染病提供适宜的技术手段，保障食品安全和人民健康。

Cholera, typhoid, dysentery and other infectious diarrhea are still main diseases of human. At present, serological detection is the gold standard of clinical identification and surveillance of enteropathogens. As we known, serological detection is based on slide agglutination test. Because of disadvantages of complexity operation, time consuming, poor repeatability and expensive expense, a new technology based on molecular identification system for serotyping which will replace the conventional one is needed urgently. With the occurrence of new technologies, several new methods have been developed in recent years. But those methods still had unacceptable defects, as listed below:(1)low detective channels; (2) contaminated PCR outcomes and complicated operation process of microarray and microspheres technology; (3) expensive equipment. Our research group has invented a novel technology named MeltPleX, which is based on several new technologies (Probe melting, multiplex real-time PCR). By using MeltPleX, 42-48 target genes can be detected in one tube on real-time PCR machine. MeltPleX would be applied to detect and classify the serotypes of 4 common enteropathogens (Cholerae,Shigella, V.P, Diarrhea-causing Escherichia coli)by using an integrated molecular identification system (extraction, amplification and detection). The whole detection of MeltPleX costs 4 hours and the result can be analyzed by software automatically. The developed method could be used in surveillance and detection of intestinal infectious diseases to ensure food safety and human health.