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.

journal_name

J Clin Microbiol

authors

Qin L,Gao Y,Ni W,Sun M,Wang Y,Yin C,Qi X,Gao H,Wang X

doi

10.1128/JCM.02030-12

subject

Has Abstract

pub_date

2013-01-01 00:00:00

pages

149-54

issue

1

eissn

0095-1137

issn

1098-660X

pii

JCM.02030-12

journal_volume

51

pub_type

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