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Vol. 45, No. 3, 2002
Issue release date: May–June 2002 (October 2002)
Intervirology 2002;45:136–141
(DOI:10.1159/000065866)

Human Rhinovirus 87 Identified as Human Enterovirus 68 by VP4-Based Molecular Diagnosis

Ishiko H.a · Miura R.a · Shimada Y.a · Hayashi A.a · Nakajima H.a · Yamazaki S.a · Takeda N.b
aResearch and Development Department, Mitsubishi Kagaku Bio-Clinical Laboratories, and bDepartment of Virology II, National Institute of Infectious Diseases, Tokyo, Japan
email Corresponding Author

Abstract

Human rhinoviruses (HRVs) are the major cause of respiratory infections. We developed a diagnostic method for HRVs based on the reverse-transcription polymerase chain reaction (RT-PCR) and VP4-based phylogenetic analysis. A set of primers used in the RT-PCR of human enteroviruses (EVs) appeared to be capable of amplifying all prototype strains of HRVs, each of which generated a 530-bp fragment. The single exception was HRV-87, which generated a 650-bp fragment, as observed in human EVs. The VP4 nucleotide sequence of HRV-87 showed more than 97% nucleotide identity with human EV-68, and formed a monophyletic cluster along with the prototype strain of EV-68 in the human EV-D cluster. HRV-87 showed the second highest homology (76.8%) with EV-70, another member of the human EV-D, in a sample of 66 human EVs and 12 HRVs. Therefore, HRV-87 should be reclassified into the cluster containing human EV-68.


 goto top of outline Key Words

  • Human rhinoviruses
  • VP4
  • RT-PCR
  • Phylogenetic analysis
  • HRV-87
  • Enterovirus 68

 goto top of outline Abstract

Human rhinoviruses (HRVs) are the major cause of respiratory infections. We developed a diagnostic method for HRVs based on the reverse-transcription polymerase chain reaction (RT-PCR) and VP4-based phylogenetic analysis. A set of primers used in the RT-PCR of human enteroviruses (EVs) appeared to be capable of amplifying all prototype strains of HRVs, each of which generated a 530-bp fragment. The single exception was HRV-87, which generated a 650-bp fragment, as observed in human EVs. The VP4 nucleotide sequence of HRV-87 showed more than 97% nucleotide identity with human EV-68, and formed a monophyletic cluster along with the prototype strain of EV-68 in the human EV-D cluster. HRV-87 showed the second highest homology (76.8%) with EV-70, another member of the human EV-D, in a sample of 66 human EVs and 12 HRVs. Therefore, HRV-87 should be reclassified into the cluster containing human EV-68.

Copyright © 2002 S. Karger AG, Basel


 goto top of outline References
  1. Ishiko H, Shimada Y, Yonaha M, Hashimoto O, Hayashi A, Sakae K, Takeda N: Molecular diagnosis of human enteroviruses by phylogeny-based classification using the VP4 sequence. J Infect Dis 2002;185:744–754.
  2. King AMQ, Brown F, Christian P, Hovi T, Hyypia T, Knowles NJ, Lemon SM, Minor PD, Palmenberg AC, Skern T, Stanway G: Picornaviridae. New York, Academic Press, 1999.
  3. Melnick JL: Enteroviruses: Polioviruses, Coxsackieviruses, Echoviruses, and Newer Enteroviruses, ed 3. Philadelphia, Lippincott-Raven, 1996.
  4. Olive DM, Al-Mufti S, Al-Mulla W, Khan MA, Pasca A, Stanway G, Al-Nakib W: Detection and differentiation of picornaviruses in clinical samples following genomic amplification. J Gen Virol 1990;71:2141–2147.
  5. Rivera VM, Welsh JD, Maizel JV Jr: Comparative sequence analysis of 5′ noncoding region of the enteroviruses and rhinoviruses. Virology 1988;165:42–50.
  6. Rotbart HA: Nucleic acid detection systems for enteroviruses. Clin Microbiol Rev 1991;4:156–168.
  7. Rotbart HA: Enteroviral infections of the central nervous system. Clin Infect Dis 1995;20:971–981.
  8. Nomoto A, Omata T, Toyoda H, Kuge S, Horie H, Kataoka Y, Genba Y, Nakano Y, Imura N: Complete nucleotide sequence of the attenuated poliovirus Sabin 1 strain genome. Proc Natl Acad Sci USA 1982;79:5793–5797.
  9. Kimura M: A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 1980;16:111–120.
  10. Saitou N, Nei M: The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406–425.
  11. Felsenstein J: Confidence limits on phylogenies: An approach using the bootstrap. Evolution 1985;39:783–791.
  12. Savolainen C, Blomqvist S, Mulders MN, Hovi T: Genetic clustering of all 102 human rhinovirus prototype strains: Serotype 87 is closed to human enterovirus 70. J Gen Virol 2002;83:333–340.
  13. Takeda N, Tanimura M, Miyamura K: Molecular evolution of the major capsid protein VP1 of enterovirus 70. J Virol 1994;68:854–862.
  14. Oberste MS, Maher K, Kilpatrick DR, Pallansch MA: Molecular evolution of the human enteroviruses: Correlation of serotype with VP1 sequence and application to picornavirus classification. J Virol 1999;73:1941–1948.

 goto top of outline Author Contacts

Hiroaki Ishiko, PhD
Research and Development Department
Mitsubishi Kagaku Bio-Clinical Laboratories Inc.
Shimura 3-30-1, Itabashi, Tokyo 174-8555 (Japan)
Tel. +81 3 5994 2332, Fax +81 3 5994 2972, E-Mail Ishiko@m1.mbcl.co.jp


 goto top of outline Article Information

Received: Received: April 22, 2002
Accepted after revision: May 1, 2002
Number of Figures : 3, Number of Tables : 0, Number of References : 14


 goto top of outline Publication Details

Intervirology (International Journal of Basic and Medical Virology)
Founded 1973 by J.L. Melnick; continued by F. Rapp (1986–1990); M.J. Buchmeier and C.R. Howard (1991–1993)

Vol. 45, No. 3, Year 2002 (Cover Date: May-June 2002 (Released October 2002))

Journal Editor: Rüdiger W. Braun, Stuttgart
ISSN: 0300–5526 (print), 1423–0100 (Online)

For additional information: http://www.karger.com/journals/int


Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Abstract

Human rhinoviruses (HRVs) are the major cause of respiratory infections. We developed a diagnostic method for HRVs based on the reverse-transcription polymerase chain reaction (RT-PCR) and VP4-based phylogenetic analysis. A set of primers used in the RT-PCR of human enteroviruses (EVs) appeared to be capable of amplifying all prototype strains of HRVs, each of which generated a 530-bp fragment. The single exception was HRV-87, which generated a 650-bp fragment, as observed in human EVs. The VP4 nucleotide sequence of HRV-87 showed more than 97% nucleotide identity with human EV-68, and formed a monophyletic cluster along with the prototype strain of EV-68 in the human EV-D cluster. HRV-87 showed the second highest homology (76.8%) with EV-70, another member of the human EV-D, in a sample of 66 human EVs and 12 HRVs. Therefore, HRV-87 should be reclassified into the cluster containing human EV-68.



 goto top of outline Author Contacts

Hiroaki Ishiko, PhD
Research and Development Department
Mitsubishi Kagaku Bio-Clinical Laboratories Inc.
Shimura 3-30-1, Itabashi, Tokyo 174-8555 (Japan)
Tel. +81 3 5994 2332, Fax +81 3 5994 2972, E-Mail Ishiko@m1.mbcl.co.jp


 goto top of outline Article Information

Received: Received: April 22, 2002
Accepted after revision: May 1, 2002
Number of Figures : 3, Number of Tables : 0, Number of References : 14


 goto top of outline Publication Details

Intervirology (International Journal of Basic and Medical Virology)
Founded 1973 by J.L. Melnick; continued by F. Rapp (1986–1990); M.J. Buchmeier and C.R. Howard (1991–1993)

Vol. 45, No. 3, Year 2002 (Cover Date: May-June 2002 (Released October 2002))

Journal Editor: Rüdiger W. Braun, Stuttgart
ISSN: 0300–5526 (print), 1423–0100 (Online)

For additional information: http://www.karger.com/journals/int


Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

References

  1. Ishiko H, Shimada Y, Yonaha M, Hashimoto O, Hayashi A, Sakae K, Takeda N: Molecular diagnosis of human enteroviruses by phylogeny-based classification using the VP4 sequence. J Infect Dis 2002;185:744–754.
  2. King AMQ, Brown F, Christian P, Hovi T, Hyypia T, Knowles NJ, Lemon SM, Minor PD, Palmenberg AC, Skern T, Stanway G: Picornaviridae. New York, Academic Press, 1999.
  3. Melnick JL: Enteroviruses: Polioviruses, Coxsackieviruses, Echoviruses, and Newer Enteroviruses, ed 3. Philadelphia, Lippincott-Raven, 1996.
  4. Olive DM, Al-Mufti S, Al-Mulla W, Khan MA, Pasca A, Stanway G, Al-Nakib W: Detection and differentiation of picornaviruses in clinical samples following genomic amplification. J Gen Virol 1990;71:2141–2147.
  5. Rivera VM, Welsh JD, Maizel JV Jr: Comparative sequence analysis of 5′ noncoding region of the enteroviruses and rhinoviruses. Virology 1988;165:42–50.
  6. Rotbart HA: Nucleic acid detection systems for enteroviruses. Clin Microbiol Rev 1991;4:156–168.
  7. Rotbart HA: Enteroviral infections of the central nervous system. Clin Infect Dis 1995;20:971–981.
  8. Nomoto A, Omata T, Toyoda H, Kuge S, Horie H, Kataoka Y, Genba Y, Nakano Y, Imura N: Complete nucleotide sequence of the attenuated poliovirus Sabin 1 strain genome. Proc Natl Acad Sci USA 1982;79:5793–5797.
  9. Kimura M: A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 1980;16:111–120.
  10. Saitou N, Nei M: The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406–425.
  11. Felsenstein J: Confidence limits on phylogenies: An approach using the bootstrap. Evolution 1985;39:783–791.
  12. Savolainen C, Blomqvist S, Mulders MN, Hovi T: Genetic clustering of all 102 human rhinovirus prototype strains: Serotype 87 is closed to human enterovirus 70. J Gen Virol 2002;83:333–340.
  13. Takeda N, Tanimura M, Miyamura K: Molecular evolution of the major capsid protein VP1 of enterovirus 70. J Virol 1994;68:854–862.
  14. Oberste MS, Maher K, Kilpatrick DR, Pallansch MA: Molecular evolution of the human enteroviruses: Correlation of serotype with VP1 sequence and application to picornavirus classification. J Virol 1999;73:1941–1948.