Sequencing analysis of whole genome of one strain of bovine Akabane virus and es-tablishment of fluorescence quantitative PCR for virus detection
10.16303/j.cnki.1005-4545.2024.09.05
- VernacularTitle:1株牛阿卡斑病毒全基因组的测定分析及荧光定量PCR检测方法的建立
- Author:
Jiafu SHANG
1
;
Le HU
;
Mingke LI
;
Yujian WU
;
Xingwei NI
;
Xiaowei YANG
;
Xia LIU
;
Liwu ZHANG
;
Tingting XU
;
Guangwei ZHAO
Author Information
1. 西南大学动物医学院,重庆荣昌 402460
- Keywords:
Akabane virus;
whole genome;
fluorescence quantitative PCR;
bovine;
Guizhou Province
- From:
Chinese Journal of Veterinary Science
2024;44(9):1872-1881
- CountryChina
- Language:Chinese
-
Abstract:
In order to understand the prevalence of Akabane disease(AKAD)in Guizhou Province and the molecular characteristics of the isolates,the whole-genome sequence of a strain of Akabane virus(AKAV)from a bovine AKAD-positive sample was determined and analyzed.The genotype and genetic variation of the strain were also explored.Based on the conserved S sequence,a fluores-cence quantitative PCR(qPCR)detection method was established and applied for the investigation of AKAV infection status in four large-scale beef cattle farms of Guizhou.Results showed that the S,M and L fragments of the bovine strain were highly homologous to the Tianjin strain(TJ2016/China/2016)and the Australian strain(JaLAB39/Australia/1959),where they were in the same evolutionary branch and belonged to genotype Ⅱ.Sensitivity assay found that the lowest detection limit was 2.5 X 101 copies/μL.Specificity assay showed the established method detected only AKAV with no amplification on bovine bluetongue virus(BLUV),Pasteurella multocida(PM),bovine infectious rhinotracheitis virus(IBRV)and bovine Mycoplasma bovis.The variation coefficients of inter-and intra batches in the repeatability test were both lower than 2.26％.These findings illus-trated that the established qPCR method had high sensitivity,good specificity and repeatability.A total of 298 serum samples from 4 large-scale beef cattle farms in Qianxi City and Huangping County of Guizhou Province were collected and tested for AKAV by the method.Out of 298 sam-ples,25 positive samples(25/298)were detected as positive with a positive rate of 8.39％.In sum-mary,this work provided the reference data for a deep understanding of the molecular prevalence of AKAV in Guizhou Province and laid foundation for the prevention and control of AKAD.
