title: Development and application of real-time PCR for detection of subgroup J avian leukosis virus.
abstract: Subgroup J avian leukosis virus (ALV-J) is an avian retrovirus that causes severe economic losses in the poultry industry. The early identification and removal of virus-shedding birds are important to reduce the spread of congenital and contact infections. In this study, a TaqMan-based real-time PCR method for the rapid detection and quantification of ALV-J with proviral DNA was developed. This method exhibited a high specificity for ALV-J. Moreover, the detection limit was as low as 10 viral DNA copies. The coefficients of variation (CVs) of both interassay and intra-assay reproducibility were less than 1%. The growth curves of ALV-J in DF-1 cells were measured by real-time PCR, yielding a trend line similar to those determined by 50% tissue culture infective dose (TCID(50)) and p27 antigen detection. Tissue samples suspected of ALV infection were evaluated using real-time PCR, virus isolation, and routine PCR, and the positivity rates were 60.1%, 41.6% and 44.5%, respectively. Our data indicated that the real-time PCR method provides a sensitive, specific, and reproducible diagnostic tool for the identification and quantification of ALV-J for clinical diagnosis and in laboratory research.
authors.affiliation: Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, People's Republic of China.
authors.forename: Liting
authors.initials: L
authors.lastname: Qin
authors.forename: Yulong
authors.initials: Y
authors.lastname: Gao
authors.forename: Wei
authors.initials: W
authors.lastname: Ni
authors.forename: Meiyu
authors.initials: M
authors.lastname: Sun
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Wang
authors.forename: Chunhong
authors.initials: C
authors.lastname: Yin
authors.forename: Xiaole
authors.initials: X
authors.lastname: Qi
authors.forename: Honglei
authors.initials: H
authors.lastname: Gao
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D001353:Avian Leukosis / Q000175:diagnosis* / Q000821:virology*; D001354:Avian Leukosis Virus / Q000235:genetics / Q000302:isolation & purification*; D005838:Genotype; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D011200:Poultry; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*; D019562:Viral Load / Q000379:methods