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通过聚合酶链反应直接检测原代粪便培养物中的大肠杆菌志贺样毒素基因。

Direct detection of Escherichia coli Shiga-like toxin genes in primary fecal cultures by polymerase chain reaction.

作者信息

Paton A W, Paton J C, Goldwater P N, Manning P A

机构信息

Department of Microbiology, Adelaide Children's Hospital, South Australia.

出版信息

J Clin Microbiol. 1993 Nov;31(11):3063-7. doi: 10.1128/jcm.31.11.3063-3067.1993.

Abstract

A single pair of oligonucleotide primers was used for polymerase chain reaction amplification of a 212- or 215-bp region of Escherichia coli Shiga-like toxin (SLT) genes from crude fecal culture extracts. Genes were typed by hybridization of the polymerase chain reaction products to SLT-I- or SLT-II-specific oligonucleotide probes. The procedure was capable of detecting fewer than 10 SLT-producing E. coli organisms per ml of culture against a background of more than 10(9) other organisms per ml and provides a rapid and sensitive means of screening primary fecal cultures for the presence of such strains. When this procedure was used to test primary cultures from gut contents or feces from various patient groups, including apparently healthy infants, approximately half of all samples yielded positive results for SLT-I and/or SLT-II sequences.

摘要

使用一对寡核苷酸引物通过聚合酶链反应从粪便粗培养提取物中扩增大肠杆菌志贺样毒素(SLT)基因的212或215bp区域。通过聚合酶链反应产物与SLT-I或SLT-II特异性寡核苷酸探针杂交对基因进行分型。该方法能够在每毫升培养物中存在超过10⁹个其他微生物的背景下,检测出每毫升培养物中少于10个产SLT的大肠杆菌,为筛查初级粪便培养物中此类菌株的存在提供了一种快速且灵敏的方法。当使用该方法检测来自包括看似健康的婴儿在内的各种患者群体的肠道内容物或粪便的初级培养物时,大约一半的样本对SLT-I和/或SLT-II序列产生阳性结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae55/266220/0860e994df9c/jcm00023-0238-a.jpg

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