期刊
JOURNAL OF VIROLOGICAL METHODS
卷 102, 期 1-2, 页码 1-8出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0166-0934(01)00372-X
关键词
QC-RT-PCR; real time RT-PCR; ALV-J; fluorogenic; probe; TCID50
Avian leukosis virus subgroup J (ALV-J) infections cause significant economic losses because mortality, tumor production, decreased production, and cost for eradication. Current quantification methods for ALV-J expressed by TCID50 are difficult to determine because of the lack of cytopathic effect in cell cultures and non-specificity of currently available antigen-capture ELISA tests. In this study, a one-tube fluorescent probe based real time RT-PCR method was developed for quantification of ALV-J and compared with available quantification methods. Cell lysates with different TCID(50)s determined by cell culture and antigen capture ELISA (ag-ELISA) were used for one-tube real time RT-PCR using fluorogenic probe and quantitative competitive RT-PCR (QC-RT-PCR). The results of QC-RT-PCR and real time RT-PCR were highly correlated to the TCID(50)s determined by conventional culture methods. They were also very specific, sensitive, easy to perform, reproducible, and rapid compared with conventional methods. These RT-PCR based quantification methods of ALV-J viral RNA will be useful for virological and pathogenesis studies, (C) 2002 Elsevier Science B.V. All rights reserved.
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