Development and Validation of DNA Microarray for Genotyping Group A Rotavirus VP4 (P[4], P[6], P[8], P[9], and P[14]) and VP7 (G1 to G6, G8 to G10, and G12) Genes
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Published:2007-08
Issue:8
Volume:45
Page:2641-2648
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ISSN:0095-1137
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Container-title:Journal of Clinical Microbiology
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language:en
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Short-container-title:J Clin Microbiol
Author:
Honma Shinjiro1, Chizhikov Vladimir2, Santos Norma13, Tatsumi Masatoshi1, do Carmo S. T. Timenetsky Maria4, Linhares Alexandre C.5, Mascarenhas Joana D'Arc P.5, Ushijima Hiroshi6, Armah George E.7, Gentsch Jon R.8, Hoshino Yasutaka1
Affiliation:
1. Epidemiology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 2. Laboratory of Method Development, Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, Maryland 3. Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janerio, Brazil 4. Instituto Adolfo Lutz, São Paulo, Brazil 5. Instituto Evandro Chagas, Secretaria de Vigilância em Saúde, Belém, Brazil 6. University of Tokyo, Tokyo, Japan 7. University of Ghana, Legon, Ghana 8. Gastroenteritis and Respiratory Viruses Laboratory Branch, Centers for Disease Control and Prevention, Atlanta, Georgia
Abstract
ABSTRACT
Previously, we reported the development of a microarray-based method for the identification of five clinically relevant G genotypes (G1 to G4 and G9) (V. Chizhikov et al., J. Clin. Microbiol. 40:2398-2407, 2002). The expanded version of the rotavirus microarray assay presented herein is capable of identifying (i) five clinically relevant human rotavirus VP4 genotypes (P[4], P[6], P[8], P[9], and P[14]) and (ii) five additional human rotavirus VP7 genotypes (G5, G6, G8, G10, and G12) on one chip. Initially, a total of 80 cell culture-adapted human and animal reference rotavirus strains of known P (P[1] to P[12], P[14], P[16], and P[20]) and G (G1-6, G8 to G12, and G14) genotypes isolated in various parts of the world were employed to evaluate the new microarray assay. All rotavirus strains bearing P[4], P[6], P[8], P[9], or P[14] and/or G1 to G6, G8 to G10, or G12 specificity were identified correctly. In addition, cross-reactivity to viruses of genotype G11, G13, or G14 or P[1] to P[3], P[5], P[7], P[10] to P[12], P[16], or P[20] was not observed. Next, we analyzed a total of 128 rotavirus-positive human stool samples collected in three countries (Brazil, Ghana, and the United States) by this assay and validated its usefulness. The results of this study showed that the assay was sensitive and specific and capable of unambiguously discriminating mixed rotavirus infections from nonspecific cross-reactivity; the inability to discriminate mixed infections from nonspecific cross-reactivity is one of the inherent shortcomings of traditional multiplex reverse transcription-PCR genotyping. Moreover, because the hybridization patterns exhibited by rotavirus strains of different genotypes can vary, this method may be ideal for analyzing the genetic polymorphisms of the VP7 or VP4 genes of rotaviruses.
Publisher
American Society for Microbiology
Subject
Microbiology (medical)
Reference60 articles.
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