Direct detection of Escherichia coli in positive blood culture bottles by loop-mediated isothermal amplification method
10.3969/j.issn.1671-8348.2015.11.025
- VernacularTitle:环介导等温扩增法直接检测阳性血培养瓶中大肠埃希菌
- Author:
Chunyang REN
;
Jie TIAN
- Publication Type:Journal Article
- Keywords:
loop-mediated isothermal amplification;
testing;
Escherichia coli
- From:
Chongqing Medicine
2015;(11):1514-1515,1518
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a rapid ,sensitive direct method for detecting Escherichia coli (E .coli) in the positive blood culture bottles by using the loop‐mediated isothermal amplification (LAMP) .Methods Based on the lacZ (accession number :M74750) gene sequence of E .coli in the gene sequence GenBank by the National Institutes of Health (NIH) ,4 specific primers (2 inner primers and outer primers) were designed ,the lacZ gene was amplified and the amplification reaction was optimized .The sen‐sitivity and specificity of this method were verified and its comparison with the traditional culture method was conducted .Results Sixty‐eight strains of E .coli were detected in 300 positive blood culture bottles by adopting the LAMP method .The designed prim‐ers had good sensitivity and good specificity for the amplification of E .coli .The sensitivity and specificity of LAMP all reached 100% compared with the traditional method ,but the traditional method needed 3 d to get the results ,while LAMP only needed 1 h . Conclusion LAMP is extremely rapid for detecting E .coli with low cost ,strong specificity and high sensitivity and is suitable for clinical application .
