Establishment and application of triplex TaqMan quantitative PCR for detection of blaNDM,mcr-1 and cfr
10.16303/j.cnki.1005-4545.2025.02.10
- VernacularTitle:耐药基因blaNDM、mcr-1和cfr三重TaqMan qPCR检测方法的建立及应用
- Author:
Wei YANG
1
;
Haihang YU
;
Yunmeng WANG
;
Jue WANG
;
Yu HAN
;
Xiaoyue HU
;
Zhiwei CHEN
;
Junxia LU
;
Ying GAO
;
Ning ZHANG
Author Information
1. 河北省畜牧兽医研究所,河北保定 071000
- Publication Type:Journal Article
- Keywords:
blaNDM;
mcr-1;
cfr;
antibiotic resistance genes;
multiplex fluorescence quantitative PCR
- From:
Chinese Journal of Veterinary Science
2025;45(2):243-248,273
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to simultaneously detect three antibiotic resistance genes(blaNDM,mcr-1 and cfr).A triplex fluorescence quantitative PCR method was established.Plasmids,primers and probes were designed and optimized.The method could specifically detect blaNDM,mcr-1 and cfr,but not other antibiotic resistance genes.The R2 of the standard curves of the three antibiotic re-sistance genes were all greater than 0.999,and the coefficients of variation were all lower than 1％.The lowest detection limits of the plasmids were 1 × 102 copies/μL.This method was used to de-tect 800 bacterial samples.The results showed that 32 samples contained mcr-1 gene,40 samples contained blaNDM gene,2 samples contained cfr gene,8 samples contained both mcr-1 and blaNDM genes.There were no samples carrying three antibiotic resistance genes detected.The results indica-ted that the triplex fluorescence quantitative PCR method established in this experiment had the advantages of high sensitivity,specificity and stability.It was suitable for rapid detection of blaNDM,mcr-1 and cfr antibiotic resistance genes in clinical practice.It provided a convenient and quick method basis for the detection of antibiotic resistance genes.
