志贺氏菌
琼脂糖凝胶电泳
大肠杆菌
生物
聚合酶链反应
微生物学
实时聚合酶链反应
分子生物学
DNA
病毒学
基因
遗传学
作者
Orntipa Sethabutr,Malabi M. Venkatesan,Sylvia Yam,Lorrin Pang,Bonnie L. Smoak,Willie Sang,P Echeverria,David N. Taylor,Daniel W. Isenbarger
标识
DOI:10.1016/s0732-8893(00)00122-x
摘要
PCR techniques applied to diarrheal stools reliably diagnose Shigella and enteroinvasive Escherichia coli (EIEC) infections. Identification of PCR products using agarose gel electrophoresis (AGE) and hybridization with DNA probes has several shortcomings. Automated methods of identifying PCR products that process larger numbers of specimens can facilitate epidemiologic studies and standardize results. In this study, we used ELISA following PCR to detect ipaH gene sequences of Shigella and EIEC from 89 diarrheal stools. Results of ELISA were compared with AGE with and without DNA probe, and with culture. Two specimen preparation methods were compared as well: boiling/centrifugation, and purification with silicon dioxide (SiO(2)). Both PCR product-detection methods identified significantly more infections than did culture. PCR-ELISA detected significantly more infections than PCR-AGE when processed using SiO2 (P = 0.014). PCR-ELISA allows screening of larger numbers of specimens, automates test results, and avoids use of mutagenic reagents. PCR-ELISA is faster than PCR-AGE when testing large numbers of specimens, although not when testing small numbers of specimens.
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