A New High-throughput Real-time PCR Assay for the Screening of Multiple Antimicrobial Resistance Genes in Broiler Fecal Samples from China.
- Author:
Jie CHE
1
;
Jin Xing LU
1
;
Wen Ge LI
1
;
Yun Fei ZHANG
1
;
Xiao Fei ZHAO
1
;
Min YUAN
1
;
Xue Mei BAI
1
;
Xia CHEN
1
;
Juan LI
1
Author Information
- Publication Type:Journal Article
- Keywords: Antimicrobial resistance; Antimicrobial resistance gene; Broiler fecal sample; High-throughput real-time PCR array
- From: Biomedical and Environmental Sciences 2019;32(12):881-892
- CountryChina
- Language:English
-
Abstract:
OBJECTIVE:Antimicrobial resistance (AMR) has become a global concern and is especially severe in China. To effectively and reliably provide AMR data, we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology, and screened multiple AMR genes in broiler fecal samples.
METHODS:A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection. A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR genes.
RESULTS:The new assay with limits of detection ranging from 40.9 to 8,000 copies/reaction. The sensitivity rate, specificity rate, positive predictive value, negative predictive value and correct indices were 99.30%, 98.08%, 95.31%, 99.79%, and 0.9755, respectively. Utilizing this assay, we demonstrate that AMR genes are widely spread, with positive detection rates ranging from 0 to 97.07% in 273 broiler fecal samples. blaCTX-M, blaTEM, mcr-1, fexA, cfr, optrA, and intI1 showed over 80% prevalence. The dissemination of AMR genes was distinct between the two farms.
CONCLUSION:We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal samples. The widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.
