Author:
Li Haiqin,Zhu Yujun,Wan Chunhe,Wang Zhangzhang,Liu Lei,Tan Meifang,Zhang Fanfan,Zeng Yanbing,Huang Jiangnan,Wu Chengcheng,Huang Yu,Kang Zhaofeng,Guo Xiaoqiao
Abstract
Abstract
Background
Goose astrovirus (GoAstV) is an important pathogen that causes joint and visceral gout in goslings. It has been circulating in many provinces of China since 2017. Goose astrovirus genotypes 2 (GoAstV-2) is the main epidemic strain, and its high morbidity and mortality have caused huge economic losses to the goose industry. An accurate point-of-care detection for GoAstV-2 is of great significance. In this study, we developed a real-time reverse transcription recombinase polymerase amplification (RT-RPA) method for the on-site detection of GoAstV-2 infection.
Results
The real-time RT-RPA reaction was carried out at a constant temperature of 39 °C, and the entire detection time from nucleic acid preparation to the end of amplification was only 25 min using the portable device. The results of a specificity analysis showed that no cross-reaction was observed with other related pathogens. The detection limit of the assay was 100 RNA copies/μL. The low coefficient of variation value indicated excellent repeatability. We used 270 clinical samples to evaluate the performance of our established method, the positive concordance rates with RT-qPCR were 99.6%, and the linear regression analysis revealed a strong correlation.
Conclusions
The established real-time RT-RPA assay showed high rapidity, specificity and sensitivity, which can be widely applied in the laboratory, field and especially in the resource-limited settings for GoAstV-2 point-of-care diagnosis.
Funder
basic research and talent training of Jiangxi Academy of Agricultural Sciences
Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention
Fujian Science and Technology Project
Jiangxi Agriculture Research System
National Natural Science Foundation
the Innovation Team of Jiangxi Agricultural University
Publisher
Springer Science and Business Media LLC
Subject
General Veterinary,General Medicine
Cited by
1 articles.
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