Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus

X.H. Wu; Z.Q. Yao; Q.Q. Zhao; S. Chen; Z.Z. Hu; Z. Xie; L.Y. Chen; J. Ji; F. Chen; X.H. Zhang; Q.M. Xie

Poultry Science (Apr 2022)

Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus

X.H. Wu,
Z.Q. Yao,
Q.Q. Zhao,
S. Chen,
Z.Z. Hu,
Z. Xie,
L.Y. Chen,
J. Ji,
F. Chen,
X.H. Zhang,
Q.M. Xie

Affiliations

X.H. Wu: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China
Z.Q. Yao: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China
Q.Q. Zhao: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China
S. Chen: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China
Z.Z. Hu: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China
Z. Xie: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China
L.Y. Chen: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China
J. Ji: Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, P. R. China
F. Chen: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou, Guangdong 510642, P. R. China
X.H. Zhang: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou, Guangdong 510642, P. R. China
Q.M. Xie: Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou, Guangdong 510642, P. R. China; Corresponding author:

Journal volume & issue: Vol. 101, no. 4
p. 101743

Abstract

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ABSTRACT: Subgroup J Avian leukosis virus (ALV-J) is an important pathogen of poultry tumor diseases. Since its discovery, it has caused significant economic losses to the poultry industry. Thus, the rapid detection of molecular level with strong specificity is particularly important whether poultry are infected with ALV-J. In this study, we designed primers and probe for real-time fluorescent reverse-transcription recombinase-aided amplification assay (RT-RAA) based on the ALV-J gp85 sequence. We had established a real-time fluorescent RT-RAA method and confirmed this system by verifying the specificity and sensitivity of the primers and probe. In addition, repeatability tests and clinical sample regression tests were used for preliminary evaluation of this detection method. The sensitivity of established method was about 101 copies/μL, and the repeatability of the CV of the CT value is 4%, indicating repeatability is good. Moreover, there was no cross-reactivity with NDV, IBV, IBDV, H9N2, MDV, and REV, and other avian leukosis virus subgroups, such as subgroups A, B, C, D, K and E. Importantly, the real-time fluorescent RT-RAA completed the test within 30 min at a constant temperature of 41°C. Forty-two clinical samples with known background were tested, and the test results were coincided with 100%. Overall, these results suggested that the real-time fluorescent RT-RAA developed in this study had strong specificity, high sensitivity, and good feasibility. The method is simple, easy, and portable, that is suitable for clinical and laboratory diagnosis, and provides technical support for the prevention and control of ALV-J.

Published in Poultry Science

ISSN: 0032-5791 (Print); 1525-3171 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Agriculture: Animal culture
Website: https://www.journals.elsevier.com/poultry-science

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