Development of a multiplex polymerase chain reaction assay for diarrheagenic Escherichia coli and Shigella spp. and its evaluation on colonies, culture broths, and stool

被引：41

作者：

Taniuchi, Mami ^{[1
]}

Walters, Carol C. ^{[2
]}

Gratz, Jean ^{[1
,3
]}

Maro, Athanasia ^{[3
]}

Kumburu, Happiness ^{[3
]}

Serichantalergs, Oralak ^{[4
]}

Sethabutr, Orntipa ^{[4
]}

Bodhidatta, Ladaporn ^{[4
]}

Kibiki, Gibson ^{[3
]}

Toney, Denise M. ^{[2
]}

Berkeley, Lynette ^{[5
]}

Nataro, James P. ^{[6
]}

Houpt, Eric R. ^{[1
]}

机构：

[1] Univ Virginia, Div Infect Dis & Int Hlth, Charlottesville, VA 22908 USA

[2] Virginia Dept Gen Serv, Div Consolidated Lab Serv, Richmond, VA 23219 USA

[3] Kilimanjaro Christian Med Ctr, Biotechnol Lab, Moshi, Tanzania

[4] Armed Forces Res Inst Med Sci, Dept Enter Dis, Bangkok 10400, Thailand

[5] Univ Maryland, Ctr Vaccine Dev, Baltimore, MD 21201 USA

[6] Univ Virginia, Dept Pediat, Charlottesville, VA 22908 USA

来源：

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE | 2012年 / 73卷 / 02期

关键词：

Multiplex PCR; Diarrheagenic E. coli; Luminex; Shigella; Enteroaggregative E. coli; Enterohemorrhagic E. coli; Enteropathogenic E. coli; Enterotoxigenic E. coli; Diarrhea; Enteroinvasive E. coli; PCR; Shiga toxin-producing E. coli; Virulence genes; YOUNG-CHILDREN; PCR; GENE; IDENTIFICATION; INFECTIONS; AMPLIFICATION; TRAVELERS; VARIANTS; O157-H7; PROBE;

D O I：

10.1016/j.diagmicrobio.2012.03.008

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

Detection of diarrheagenic Escherichia coli (DEC) typically depends on identification of virulence genes from stool cultures, not on stool itself. We developed a multiplex polymerase chain reaction (PCR) assay that detects key DEC virulence genes (stx1, stx2, eae, bfpA, ipaH, LT, STh, aaiC, aatA). The assay involved a multiplex PCR reaction followed by detection of amplicon(s) using Luminex beads. The assay was evaluated on over 100 colony and broth specimens. We then evaluated the assay using DNA extracted from stool, colony pools, and Gram-negative broths, using stool spiked with known quantities of DEC. Performance of the assay on stool DNA was most quantitative, while stool broth DNA offered the lowest limit of detection. The assay was prospectively evaluated on clinical specimens in Tanzania. Stool DNA yielded higher sensitivity than colony pools compared with broth DNA as the standard. We propose using this assay to screen for DEC directly in stool or stool broths. (C) 2012 Elsevier Inc. All rights reserved.

引用

页码：121 / 128

页数：8

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