Quantification of chicken anaemia virus by competitive polymerase chain reaction

A quantitative method for chicken anaemia virus (CAV) was developed using competitive polymerase chain reaction (PCR). Competitive template was constructed by deletion of 33 nucleotides from a wildtype DNA clone of CAV. Quantification of CAV DNA molecules by the competitive PCR was rapid and highly reproducible when compared with conventional infectivity titration methods. The ratios of the viral DNA molecules and infectivity titres in MDCC-MSB1 cells varied between 1.3 and 3.55 log10 among several isolates, suggesting the existence of different infection efficiencies to MDCC-MSB1 cells by isolates. The competitive PCR will be useful for studying CAV infection in vivo and/or in vitro.

[1]  Kanji Hirai,et al.  Marek’s Disease , 2001, Current Topics in Microbiology and Immunology.

[2]  K. Nakamura,et al.  Detection of chicken anaemia virus DNA from formalin-fixed tissues by polymerase chain reaction. , 1998, Research in veterinary science.

[3]  A. Tanaka,et al.  Cloning and sequencing of the genome of chicken anaemia virus (CAV) TK-5803 strain and comparison with other CAV strains. , 1996, The Journal of veterinary medical science.

[4]  S. Takagi,et al.  Pathogenicity for chicks of six isolates of chicken anemia virus (CAV) isolated in the past two years ('93, '94) , 1996 .

[5]  C. Parrish,et al.  Determination of the detection limit of the polymerase chain reaction for chicken infectious anemia virus. , 1993, Avian diseases.

[6]  K. Mawhinney,et al.  Detection and differentiation of chicken anemia virus isolates by using the polymerase chain reaction , 1992, Journal of clinical microbiology.

[7]  M. Noteborn,et al.  Characterization of cloned chicken anemia virus DNA that contains all elements for the infectious replication cycle , 1991, Journal of virology.

[8]  M. Mcnulty.  Chicken anaemia agent: a review. , 1991, Avian pathology : journal of the W.V.P.A.

[9]  M. Mcnulty,et al.  Economic effects of subclinical chicken anemia agent infection in broiler chickens. , 1991, Avian diseases.

[10]  K. Imai,et al.  Development of a microtest method for serological and virological examinations of chicken anemia agent. , 1990, Nihon juigaku zasshi. The Japanese journal of veterinary science.

[11]  F. Rixon,et al.  Purification and biochemical characterization of chicken anaemia agent. , 1990, The Journal of general virology.

[12]  T. Nunoya,et al.  Isolation of chicken anaemia agent and Marek's disease virus from chickens vaccinated with turkey herpesvirus and lesions induced in chicks by inoculating both agents. , 1987, Avian pathology : journal of the W.V.P.A.

[13]  K. Imai,et al.  Pathogenicity and antigenicity of eleven isolates of chicken anaemia agent (CAA). , 1986, Avian pathology : journal of the W.V.P.A.

[14]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[15]  N. Yuasa.  Propagation and infectivity titration of the Gifu-1 strain of chicken anemia agent in a cell line (MDCC-MSB1) derived from Marek's disease lymphoma. , 1983, National Institute of Animal Health quarterly.

[16]  T. Taniguchi,et al.  Isolation and some characteristics of an agent inducing anemia in chicks. , 1979 .

[17]  Y. Akiyama,et al.  Two cell lines from lymphomas of Marek's disease. , 1974, Biken journal.