中国兽医科学2024，Vol.54Issue(12)：1577-1585,9.DOI:10.16656/j.issn.1673-4696.2024.0220
圆圈病毒禽源1型实时荧光RAA快速检测方法的建立
Establishment of the real-time RAA detection method for Gyrovirus galga 1
摘要
Abstract
In order to establish a rapid detection method for Gyrovirus galga 1(GyVg1),the specific primers and probe were designed based on the conserved region of GyVg1 by searching and comparing se-quences currently available in GenBank.After screening primers and optimization of the reaction con-ditions,a recombinase acid amplication(RAA)detection method for GyVg1 was established.The results showed that GyVg1 could be identified in 20 minutes by the real-time fluorescence RAA assay.This method could only distinguish GyVg1,but not react with Gyrovirus homsa1(GyH1),chicken infectious anemia virus(CIAV),fowl adenovirus 4(FAdV4),H9 subtype avian influenza virus(H9-AIV),Newcastle disease virus(NDV),avian infectious bronchitis virus(IBV),infectious bursal disease virus(IBDV),chicken parvovirus(ChPV),avian leukosis virus(ALV),Mycoplasma gallisepticum(MG)and Mycoplasma synoviae(MS);The lowest limit of detection was 1 × 102 copies/μL and the coefficient of variations in intra-and inter-assay were both less than 6％.231 samples were tested simultaneously by fluorescence RAA and fluorescence quantitative PCR,and the results were consistent with each other.In conclusion,a re-al-time fluorescence RAA method is successfully established for the first time and this method has good specificity,sensitivity and repeatability,which provides a new technical means for the accurate and rapid detection of GyVg1 infection.关键词
圆圈病毒禽源1型/重组酶介导核酸等温扩增/核酸检测Key words
Gyrovirus galga 1/recombinase acid amplication/nucleic acid detection分类
农业科技引用本文复制引用
余丹,赵俊柯,喻华英,张艳芳,尹文巧,谢志勤,李小凤,吴嫒琼,谢芝勋..圆圈病毒禽源1型实时荧光RAA快速检测方法的建立[J].中国兽医科学,2024,54(12):1577-1585,9.基金项目
广西科技基地和人才专项(桂科AD17195083) （桂科AD17195083）
广西兽医生物技术重点实验室自主研究项目(24-035-32-A-01) （24-035-32-A-01）
广西科技先锋队项目(202409-01) （202409-01）
广西八桂学者专项(2019A50) （2019A50）
