Establishment and Application of a Real-Time Recombinase Polymerase Amplification Assay for the Detection of Avian Leukosis Virus Subgroup J

• DOI: 10.3389/fvets.2022.847194
• Publication Date: 2022-07-07T08:42:40Z
• Authors: Guanggang Qu; Yun Li; Zhongwei Zhao; Lizhong Miao; Feng Wei; Na Tang; Qingqing Xu; Venugopal Nair; Yongxiu Yao; Zhiqiang Shen
• Keywords: Avian leukosis; Recombinase Polymerase Amplification

Avian leukosis caused by avian virus (ALV), belonging to the genus Alpharetrovirus of family Retroviridae, is associated with benign and malignant tumors in hemopoietic cells poultry. Although several methods have been developed for ALV detection, most them are not suitable rapid on-site testing due instrument limitations, professional operators, or low sensitivity method. Herein, we described real-time recombinase polymerase amplification (RPA) assay detection subgroup J (ALV-J). The major viral structural glycoprotein gp85, highly specific subgroup, was used as molecular target RPA assay. results were obtained at 38°C within 20 min, 10 copies/μl standard plasmid pMD18-T-gp85 template per reaction. Real-time capable ALV-J-specific without cross-reaction other non-targeted pathogens. Of 62 clinical samples tested, ALV-positive rates RPA, PCR, PCR 66.13% (41/62), 59.68% (37/62), 67.74% (42/62), respectively. diagnostic agreement between 98.39% (61/62), kappa value 0.9636. ALV-J seems promising sensitive laboratories. It especially a poor resource environment, thus facilitating prevention control ALV-J.