Rapid detection of Enteroinvasive Escherichia coli by nucleic acid immunochromatography

Abstract

ObjectiveTo establish a nucleic acid immunochromatography method for rapid detection of Enteroinvasive Escherichia coli （EIEC）.MethodsThe target single-stranded DNA was prepared by asymmetric PCR， the invE which was the virulence gene of EIEC and uidA which was the marker gene of Escherichia coli were detected by immunochromatography.ResultsIn the asymmetric PCR system， the optimal ratio of forward and reverse primer for uidA and invE was 1∶3， and the optimal primer concentrations （reverse primer） were 0.2 μmol/L and 0.25 μmol/L， respectively. The optimal cycle number of amplification was 40. The lowest detection limit of this method was 3.97 × 10-3 ng/μL， and the specificity was comparable to that of PCR-gel electrophoresis.ConclusionThe method which had the advantages of convenience， less time consuming， low detection cost， good accuracy and specificity， was suitable for primary laboratory.

Keywords

• detection
• enteroinvasive escherichia coli
• virulence gene
• immunochromatography