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Applied and Environmental Microbiology, February 2004, p. 1059-1067, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1059-1067.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Restriction Fragment Length Polymorphisms and Sequence Analysis: an Approach for Genotyping Infectious Pancreatic Necrosis Virus Reference Strains and Other Aquabirnaviruses Isolated from Northwestern Spain{dagger}

J. M. Cutrín,1* J. L. Barja,1 B. L. Nicholson,2 I. Bandín,1 S. Blake,2 and C. P. Dopazo1

Unidad de Ictiopatología, Instituto de Acuicultura, Departamento de Microbiología y Parasitología, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain,1 Department of Biochemistry, Microbiology and Molecular Biology, University of Maine, Orono, Maine 044692

Received 5 August 2003/ Accepted 12 November 2003

Reference strains of infectious pancreatic necrosis virus resembling the 10 recognized serotypes and local isolates of aquabirnaviruses isolated in northwestern Spain from reservoirs (mollusks) and from asymptomatic and carrier cultured fish were genotyped by restriction fragment length polymorphism (RFLP) and nucleic acid sequence analyses. The RFLP analysis yielded seven genogroups, each of which was clearly correlated with a serotype. Sequence analysis of the three open reading frames provided quite similar results in terms of genogrouping. Based on the results of this study and in order to unify the two types of assays, we propose placing aquabirnaviruses into six genogroups, four of which can be subdivided into two genotypes based on a two-step restriction analysis. The genotyping corresponds with serotyping as follows: genogroup I includes two genotypes corresponding to serotypes A9 (genotype I.1) and A1 (genotype I.2); genogroup II corresponds to serotype A3; genogroup III includes genotypes III.1 (serotype A2) and III.2 (serotype B1); genogroups IV and V include two genotypes, each corresponding to serotypes A5, A6, A7, and A8 (genotypes IV.1, IV.2, V.1, and V.2, respectively);and genogroup VI corresponds to serotype A4. As expected, most local isolates belonged to genotype III.1 and genogroup II. However, a few local isolates corresponded to the American types of genogroup I. Finally, based on the results of this study and due to its simplicity, the two-step restriction analysis assay is proposed as a method for typing new isolates of aquabirnaviruses, and the results correspond to the results of conventional serotyping.

* Corresponding author. Mailing address: Unidad de Ictiopatología, Instituto de Acuicultura, Departamento de Microbiología y Parasitología, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain. Phone: 34 981 563 100, ext. 16052. Fax: 34 981 547 165. E-mail: iacuvir{at}uscmail.usc.es.

{dagger} Scientific contribution no. 001/2003 of the Instituto de Acuicultura.

Applied and Environmental Microbiology, February 2004, p. 1059-1067, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1059-1067.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Nishizawa, T., Kinoshita, S., Yoshimizu, M. (2005). An approach for genogrouping of Japanese isolates of aquabirnaviruses in a new genogroup, VII, based on the VP2/NS junction region. J. Gen. Virol. 86: 1973-1978 [Abstract] [Full Text]  


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