PCR快速检测腹泻患者粪中痢疾杆菌致病基因ipaH和ial
痢疾杆菌;粪便;聚合酶链反应;ipaH基因;ial基因,李小丽,阴赪宏,温艳,黄敏君,栗绍刚,齐志群,李威,通讯作者:,RapidpolymerasechainreactionassayforipaHandialgenesinfecesf
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李小丽, 阴赪宏, 温艳, 黄敏君, 栗绍刚, 齐志群, 李威, 北京热带医学研究所 首都医科大学附属北京友谊医院 北京市 100050
李小丽, 2005级首都医科大学附属北京友谊医院硕士生, 主要从事细菌性痢疾的研究.
通讯作者: 阴赪宏, 100050, 北京市宣武区永安路95号, 首都医科大学附属北京友谊医院北京热带医学研究所.
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收稿日期: 2007-04-27 接受日期: 2007-05-24
Rapid polymerase chain reaction assay for ipaH and ial genes in feces from shigellosis
Xiao-Li Li, Cheng-Hong Yin, Yan Wen, Min-Jun Huang, Shao-Gang Li, Zhi-Qun Qi, Wei Li
Xiao-Li Li, Cheng-Hong Yin, Yan Wen, Min-Jun Huang, Shao-Gang Li, Zhi-Qun Qi, Wei Li, Beijing Tropical Medicine Institute, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
Correspondence to: Cheng-Hong Yin, Beijing Tropical Medicine Institute, Beijing Friendship Hospital, Capital Medical University, 95 Yong’an Road, XuanWu District, Beijing 100050, China. modscn@yahoo.com.cn
Received: 2007-04-27 Accepted: 2007-05-24
Abstract
AIM: To establish a fast, specific and sensitive diagnostic method for detecting the invasion plasmid antigen H (ipaH) and the invasion-associated locus (ial) DNA sequence of Shigella.
METHODS: Seventy-one stool specimens from patients with acute diarrhea were tested by two methods; a traditional culture method, and a polymerase chain reaction (PCR) and nested-PCR assay. PCR–amplified products were initially evaluated by electrophoresis with 20 g/L agarose gel and visualization by ethidium bromide staining.
RESULTS: Serological assay classified the Shigella isolates as follows: 24 Shigella flexneri, 23 Shigella sonnei and one Shigella dysenteriae. Salmonella was isolated from 23 of the 71 stool specimens by conventional coproculture. PCR assays showed that 46 of the 48 Shigella spp. were positive for theipaH gene, 34 were positive for the ial gene, and 14 were negative. Nested-PCR assays detected 14 specimens as negative for ial, 12 were positive, and 46 specimens were positive for both the ipaH and ial genes ......
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