Establishment of duplex quantitative real-time PCR detection method for gyrovirus galga1 and gyrovirus homsa1
10.16303/j.cnki.1005-4545.2025.01.09
- VernacularTitle:圆圈病毒禽源1型和圆圈病毒人源1型二重荧光定量PCR检测方法的建立
- Author:
Dan YU
1
;
Zhixun XIE
;
Junke ZHAO
;
Yanfang ZHANG
;
Zhiqin XIE
;
Liji XIE
;
Wen-qiao YIN
;
Huaying YU
Author Information
1. 广西大学动物科学技术学院,广西南宁 530004;广西壮族自治区兽医研究所农业农村部中国(广西)-东盟跨境动物疫病防控重点实验室/广西兽医生物技术重点实验室,广西南宁 530001
- Publication Type:Journal Article
- Keywords:
gyrovirus galga1;
gyrovirus homsa1;
duplex real-time PCR
- From:
Chinese Journal of Veterinary Science
2025;45(1):59-65,73
- CountryChina
- Language:Chinese
-
Abstract:
Gyrovirus galga1(GyVg1)and gyrovirus homsa1(GyH1)are two newly discovered cir-coviruses that can cause symptoms related to transmissible viral proventriculitis of chickens.These viruses have been reported in various regions worldwide.This research aims to establish a duplex real-time PCR assay capable of identifying and detecting GyVg1 and GyH1.Specific primers and probes were designed based on the conserved regions of GyVg1 and GyH1 respectively using all genome sequence data currently available in GenBank.After optimizing reaction conditions,the du-plex real-time PCR detection method was established and further validated by comparing it with a conventional PCR assay and sequencing results from an analysis of 256 clinical samples collected in 2023 across eight regions of Nanning,Guangxi.The results showed that GyVg1 and GyH1 could be identified in 1 h by the duplex real-time PCR assay and two pairs of primer probes can amplify effectively but there is no any cross reaction with other pathogens.Besides,the detection limit was determined to be 7.5 copies/μL.The correlation coefficient of standard curves exceeded 0.99,and CV for intra-and inter-assay was less than 0.45％.Based on clinical performance,when the quanti-ty of template was greater than or equal to 100 copies,the agreements between the duplex real-time PCR assay and the conventional PCR assay were 94.3％(GyVg1)and 100％(GyH1).In con-clusion,the newly developed duplex real-time PCR assays exhibited good specificity,sensitivity and repeatability,which could contribute to the rapid detection and differentiation of GyVg1 and GyH1.
