大肠埃希菌O157：H7嵌合荧光RAM检测分析

赵春燕; 李凡; 刘晓梅

doi:10.11847/zgggws2006-22-06-19

大肠埃希菌O157：H7嵌合荧光RAM检测分析

Detection of Escherichia coli O157:H7 by SYBR Green I real-time ramification amplification method

摘要

摘要:
目的建立大肠埃希菌O157:H7的嵌合荧光法(SYBRGreen I)实时网状分枝扩增(RAM)检测技术.
方法将产志贺样毒素大肠埃希菌O157:H7靶基因递比稀释确定SYBRGreen I实时RAM的灵敏度, 并进一步检测临床分离的菌株.
结果 SYBRGreen I实时网状分枝扩增技术最低能检测10个产志贺样毒素大肠埃希菌O157:H7, 检测信号出现的时间与靶基因的浓度成正比, 临床分离3株产志贺样毒素大肠埃希菌O157为阳性, 而非致病性大肠埃希菌为阴性.
结论 SYBRGreen I实时RAM是一种快速、灵敏、准确、实时、环保的检测大肠埃希菌O157:H7的新核酸扩增技术.

Abstract:
Objective To establish SYBRGreen I real-time ramification amplification method(RAM)fordetection of Escherichia coli O157:H7.
Methods The bacteria were diluted in 100-fold serial with saline from 107 to 10 bacteria/Ll, and detected by rea-l time RAMfordetermining the sensitivity.Clinical isolates were detected forfurtherstudy.
Results The lowest numberof targets detected by rea-l time RAMwas 10 bacteria/Ll, and the time of appearance of detectable signal was depended on the target concentration.Three clinical isolates of E.coli O157:H7 were positive forshig a tox in gene, while nonpathogenic E.coli were negative.
Conclusion SYBRGreen Ireal-time RAMis rapid, accurate, simple, real-time environ-mental protecting, and could be another DNA amplification method to detect E.coli O157:H7.

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