Houng H S, Sethabutr O, Echeverria P
Department of Enteric Infections, Walter Reed Army Institute of Research, Washington, DC 20307-5100, USA.
Diagn Microbiol Infect Dis. 1997 May;28(1):19-25. doi: 10.1016/s0732-8893(97)89154-7.
A simple polymerase chain reaction (PCR) procedure using IS630-specific primers was developed as a general diagnostic probe to detect Shigella and enteroinvasive Escherichia coli (EIEC). However, IS630 and the other two previously reported molecular probes, ipaH and ial, cannot be used to differentiate among Shigella serotypes and EIEC strains that cause dysentery. The sensitivity of PCR protocol was determined to be 100-200 shigellae for each PCR reaction. An enrichment incubation would allow the detection of shigellae in stool samples with low bacterial concentration; i.e., < 10(4) CFU/gram. Serotype-specific primers derived from the rfc genes of differentiate among Shigella serotypes in the laboratory, such as S. sonnei, S. flexneri, and S. dysenteriae 1. It was demonstrated further that the multiplex PCR system containing rfc-specific primers can efficiently identify the most prominent Shigella serotypes in raw stool samples of acute diarrheal patients.
开发了一种使用IS630特异性引物的简单聚合酶链反应（PCR）程序，作为检测志贺氏菌和肠侵袭性大肠杆菌（EIEC）的通用诊断探针。然而，IS630以及其他两种先前报道的分子探针ipaH和ial，不能用于区分引起痢疾的志贺氏菌血清型和EIEC菌株。每个PCR反应的PCR方案灵敏度确定为100 - 200个志贺氏菌。富集培养可以检测细菌浓度低的粪便样本中的志贺氏菌；即<10(4) CFU/克。源自rfc基因的血清型特异性引物可在实验室中区分志贺氏菌血清型，如宋内志贺氏菌、福氏志贺氏菌和痢疾志贺氏菌1型。进一步证明，包含rfc特异性引物的多重PCR系统可以有效地识别急性腹泻患者原始粪便样本中最主要的志贺氏菌血清型。
