副溶血性弧菌tdh基因的分子信标PCR技术检测

万成松; 谭翰清; 温文川

副溶血性弧菌tdh基因的分子信标PCR技术检测

Detection of thermostable direct hemolysin gene in Vibrio parahaemolyticus by molecular beacon PCR

摘要

摘要:
目的采用分子信标PCR技术进行副溶血性弧菌tdh基因检测。
方法在反应体系中加入分子信标探针, 对13株副溶血性弧菌和其他细菌分别进行tdh基因实时和终点法荧光检测。
结果 2株副溶血性弧菌和阳性质粒的终点法检测荧光值分别为114.9, 95.2, 90.0。实时PCR检测的CT值分别为26.2, 26.8, 32.0。其他肠道细菌终点法检测荧光值为47.0~69.1;CT值> 32.0或无值, 与琼脂糖电泳分析结果一致。
结论分子信标PCR技术可以准确、快速、实时、简便地进行副溶血性弧菌tdh基因检测。

Abstract:
Objective To detect the thermostable direct hemolysin(tdh)gene of Vibrio parahaemolyticus using molecular beacon PCR.
Methods The tdh gene of 2 strains of Vibrio parahaemolyticus and 13 strains of other enteric pathogens was respectively amplified in the PCR reaction system containing a molecular beacon probe, and record the fluorescent value in the terminal of PCR and during the real time PCR.
Results The fluorescent value of 2 strains of Vibrio parahaemolyticus and the positive plasmid were 114.9, 95.2, 90.0 when reading in the terminal of PCR, others were 47.0~69.1;and the Ct value of 2 strains of V. parahaemolyticus and the positive plasmid were 26.2, 26.8, 32.0 using molecular beacon real time PCR, and others were over 32.The results using PCR were identical with the agarose gel electrophrosis.
Conclusion Molecular beacon PCR is a rapid, special, sensitive, convenient technique to detect the V. parahaemolyticus possessing tdh gene.

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