A Multiplex Quantitative Polymerase Chain Reaction for the Rapid Differential Detection of Subgroups A, B, J, and K Avian Leukosis Viruses

Junfeng Dou; Zui Wang; Li Li; Qin Lu; Xinxin Jin; Xiaochun Ling; Zhengyu Cheng; Tengfei Zhang; Huabin Shao; Xinguo Zhai; Qingping Luo

doi:10.3390/v15091789

A Multiplex Quantitative Polymerase Chain Reaction for the Rapid Differential Detection of Subgroups A, B, J, and K Avian Leukosis Viruses

Viruses. 2023 Aug 23;15(9):1789. doi: 10.3390/v15091789.

Authors

Junfeng Dou^{1

2

3}, Zui Wang^{1

2}, Li Li^{1

2}, Qin Lu^{1

2}, Xinxin Jin^{1

2}, Xiaochun Ling^{1

2}, Zhengyu Cheng^{1

2

4}, Tengfei Zhang^{1

2}, Huabin Shao^{1

2}, Xinguo Zhai³, Qingping Luo^{1

2}

Affiliations

¹ Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture and Rural Affairs), Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Special One, Nanhuyaoyuan, Hongshan District, Wuhan 430064, China.
² Hubei Hongshan Laboratory, Wuhan 430064, China.
³ Department of Animal Medicine, College of Life Science and Food Engineering, Hebei University of Engineering, Handan 056038, China.
⁴ Department of Microbiology and Immunology, Dalhousie University, Halifax, NS B3H 4R2, Canada.

Abstract

Avian leukosis (AL), caused by avian leukosis virus (ALV), is a contagious tumor disease that results in significant economic losses for the poultry industry. Currently, ALV-A, B, J, and K subgroups are the most common in commercial poultry and cause possible coinfections. Therefore, close monitoring is necessary to avoid greater economic losses. In this study, a novel multiplex quantitative polymerase chain reaction (qPCR) assay was developed to detect ALV-A, ALV-B, ALV-J, and ALV-K with limits of detection of 40, 11, 13.7, and 96 copies/µL, respectively, and no cross-reactivity with other ALV subtypes and avian pathogens. We detected 852 cell cultures inoculated with clinical samples using this method, showing good consistency with conventional PCR and ELISA. The most prevalent ALV strain in Hubei Province, China, was still ALV-J (11.74%). Although single infections with ALV-A, ALV-B, and ALV-K were not found, coinfections with different subgroup strains were identified: 0.7% for ALV-A/J, 0.35% for ALV-B/J, 0.25% for ALV-J/K, and 0.12% for ALV-A/B/K and ALV-A/B/J. Therefore, our novel multiplex qPCR may be a useful tool for molecular epidemiology, clinical detection of ALV, and ALV eradication programs.

Keywords: avian leukosis virus; multiplex quantitative PCR; subgroup A/B/J/K.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Avian Leukosis Virus* / genetics
Avian Leukosis* / diagnosis
Cell Culture Techniques
Coinfection* / diagnosis
Coinfection* / veterinary
Multiplex Polymerase Chain Reaction

Grants and funding

This study was funded by research on technological innovation of prevention and control of genetic diseases (G2022027013L), the China Agriculture Research System of MOF and MARA (CARS-41-G13), and Hubei Major Science and Technology Research and Development Project (2020-620-000-002-04 and 2020-620-000-002-01).