Establishment and application of CODEHOP PCR assay for detection of Pasteurella spp.in laboratory animals
10.3969.j.issn.1671-7856.2017.01.017
- VernacularTitle:巴斯德杆菌属CODEHOP PCR检测方法的建立与初步应用
- Author:
Jin XING
;
Yufang FENG
;
Bingfei YUE
;
Zhengming HE
;
Xiaomei SUN
;
Jiejie DAI
- Keywords:
Pasteurella spp.;
CODEHOP;
PCR detection;
Laboratory animals
- From:
Chinese Journal of Comparative Medicine
2017;27(1):85-90
- CountryChina
- Language:Chinese
-
Abstract:
Objective We established a rapid detection method of Pasteurella spp.and provided a reference for microbiological quality control of laboratory animal .Methods According to the β subunit of bacterial RNA polymerase ( rpoB) protein multiple alignments of 13 different Pasteurella spp.published in NCBI .The degenerate primers were designed by CODEHOP designer online .CODEHOP PCR method was applied to detecting Pasteurella spp.after the specificity and sensitivity of the method had been evaluated by 21 reference strains .Results Standard strain amplified fragment were about 200 bp by degenerate primers PastF6/PastR5.The primers are able to distinguish between Pasteurella spp.and the other pathognic organisms of laboratory animal respiratory tracts .Sensitivity of this method were 0.2 pg/μL~2 pg/μL to different Pasteurella.The Pasteurella positive rate was 19.1% in 609 animal ' s respiratory samples .The accuracy of positive results was 100%through verifying by sequenced and blast .Conclusions The established method has good specificity and sensitivity .It can be used to detect Pasteurella spp.in animal samples .
