利用表面等离子体共振（SPR）检测人粪便样本中大肠杆菌O157:H7的PCR产物。

Detection of PCR products of Escherichia coli O157:H7 in human stool samples using surface plasmon resonance (SPR).

作者信息

Kai E, Ikebukuro K, Hoshina S, Watanabe H, Karube I

机构信息

Research Center for Advanced Science and Technology, The University of Tokyo, Komaba, Japan.

出版信息

FEMS Immunol Med Microbiol. 2000 Dec;29(4):283-8. doi: 10.1111/j.1574-695X.2000.tb01535.x.

DOI:10.1111/j.1574-695X.2000.tb01535.x

PMID:11118909

Abstract

A method for the rapid detection of verotoxin-producing Escherichia coli O157:H7 in stools was evaluated. Strains possessing Shiga toxin-2 (stx-2) genes were isolated from stool samples and amplified using oligonucleotide primers. Stools spiked with cultured E. coli O157:H7 (strain 298 or strain 1646) were detected to be polymerase chain reaction (PCR) positive at 10(2) cfu per 0.1 g of stool. Stool samples from patients and healthy carriers showed a high correlation between positive results for a PCR and the presence of verotoxin-producing E. coli O157:H7, confirmed by isolation of serotype O157:H7 on sorbitol MacConkey medium (10 of 10 stool samples). These PCR products could be detected using a BIAcore 2000 surface plasmon resonance device using peptide nucleic acid as a sensor probe. In this report we use this method for the rapid detection of DNA from significant pathogenic organisms.

摘要

对一种用于快速检测粪便中产志贺毒素大肠杆菌O157:H7的方法进行了评估。从粪便样本中分离出携带志贺毒素2（stx-2）基因的菌株，并使用寡核苷酸引物进行扩增。接种培养的大肠杆菌O157:H7（菌株298或菌株1646）的粪便，每0.1克粪便中含有10² 菌落形成单位（cfu）时，检测到聚合酶链反应（PCR）呈阳性。患者和健康携带者的粪便样本显示，PCR阳性结果与产志贺毒素大肠杆菌O157:H7的存在之间具有高度相关性，这通过在山梨醇麦康凯培养基上分离出O157:H7血清型得到证实（10份粪便样本中有10份）。这些PCR产物可以使用BIAcore 2000表面等离子体共振装置，以肽核酸作为传感器探针进行检测。在本报告中，我们使用这种方法快速检测重要致病生物体的DNA。