Skip to main content
Wiley - PMC COVID-19 Collection logoLink to Wiley - PMC COVID-19 Collection
. 2005 Dec 7;44(4):323–329. doi: 10.1002/jmv.1890440403

Polymerase chain reaction and sequencing for typing rhinovirus RNA

Julie Mori 1,2, Jonathan P Clewley 1,
PMCID: PMC7166759  PMID: 7897363

Abstract

Primers were designed and tested for their ability to distinguish rhinoviruses from enterovi‐ruses. A primer set derived from the 5′‐UTR/VP coding region junction was able to amplify all the rhinovirus serotypes tested. Enteroviruses were either not amplified by these primer pairs or produced a band of larger size that could easily be discriminated from the rhinovirus‐specific product. In contrast, primers embedded in the 5′‐UTR region alone were able to amplify both rhinovirus and enterovirus RNA. It is shown that rhino‐viruses could be specifically typed by sequencing the amplicon derived from this 5′‐UTR set. The sequences of the 5′‐UTR region often previously unsequenced rhinoviruses were derived. The sequences obtained cluster into two groups: 18,41, 15, 30, 63, 31,56, and 44; and 17, 69, and 70. Ampliconsfrom serotypes 17, 69, and 70 also group by sequence with the equivalent region of HRV14 from the genetic database, while the others group with 2 and 89. © 1994 Wiley‐Liss, inc.

Keywords: enteroviruses, picornaviruses, 5′‐UTR‐VP coding region

References

  1. Agol VI (1991): The 5′‐untranslated region of picornaviral genomes. Advancesin Virus Research 40: 103–180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arruda E, Hayden FG (1993): Detection of human rhinovirus RNA in nasal washings. Molecular and Cellular Probes 7: 373–379. [DOI] [PubMed] [Google Scholar]
  3. Balfour‐Lynn IM, Valman HB, Stanway G, Khan M (1992): Use of the polymerase chain reaction to detect rhinovirus in wheezy infants. Archives of Disease in Childhood 67: 760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Becker Y, Darai G, ed (1992): “Diagnosis of Human Viruses by Polymerase Chain Reaction Technology.” Berlin: Springer‐Verlag. [Google Scholar]
  5. Boom R, Sol CJA, Salimans MMM, Jansen CL, Wertheim‐van Dillen PME, van der Noorda J (1990): Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology 28: 495–503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bruce CB, Gama RE, Hughes PJ, Stanway G (1990): A novel method of typing rhinoviruses using the product of a polymerase chain reaction. Archives of Virology 113: 83–87. [DOI] [PubMed] [Google Scholar]
  7. Corpet F (1988): Multiple sequence alignment with hierarchical clustering. Nucleic Acids Research 16: 10881–10890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Couch RB (1992): Rhinoviruses In Lennnette EH. (ed): “Laboratory Diagnosis of Viral Infections.” New York: Marcel Dekker, Inc., pp 709–729. [Google Scholar]
  9. Gama RE, Hughes PJ, Bruce CB, Stanway G (1988): Polymerase chain reaction amplification of rhinovirus nucleic acids from clinical material. Nucleic Acids Research 16: 9346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gama RE, Horsnell PR, Hughes PJ, North C, Bruce CB, Al‐Nakib W, Stanway G (1989): Amplification of rhinovirus specific nucleic acids from clinical samples using the polymerase chain reaction. Journal of Medical Virology 28: 73–77. [DOI] [PubMed] [Google Scholar]
  11. Gibson KM, Mori J, Clewley JP (1993): Detection of HIV‐1 in serum, using reverse transcription and the polymerase chain reaction (RT‐PCR). Journal of Virological Methods 43: 101–111. [DOI] [PubMed] [Google Scholar]
  12. Gow JW, Behan WMH, Clements GB, Woodall C, Riding M, Behan PO (1991): Enteroviral RNA sequences detected by polymerase chain reaction in muscle of patients with postviral fatigue syndrome. British Medical Journal 302: 692–696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gwaltney JM, Colonno RJ, Hamparian VV, Turner RB (1989): Rhino‐viruses In Schmidt NJ, Emmons RW. (eds): “Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections.” Washington, D.C.: American Public Health Association, Inc., pp 579–614. [Google Scholar]
  14. Hughes PJ, North C, Jellis CH, Minor PD, Stanway G (1988): The nucleotide sequence of human rhinovirus IB: Molecular relationships within the rhinovirus genus. Journal of General Virology 69: 49–58. [DOI] [PubMed] [Google Scholar]
  15. Hyypia T, Auvinen P, Maaronen M (1989): Polymerase chain reaction for human picornaviruses. Journal of General Virology 70: 3261–3268. [DOI] [PubMed] [Google Scholar]
  16. Jang SK, Pestova TV, Hellen CU, Witherell GW, Wimmer E (1990): Cap‐independent translation of picornavirus RNAs: Structure and function of the internal ribosomal entry site. Enzyme 44: 292–309. [DOI] [PubMed] [Google Scholar]
  17. Olive DM, Al‐Mufti S, Al‐Mulla W, Khan MA, Pasca A, Stanway G, Al‐Nakib W (1990): Detection and differentiation of picornaviruses in clinical samples following genomic amplification. Journal of General Virology 71: 2141–2147. [DOI] [PubMed] [Google Scholar]
  18. Persing DH, Smith TF, Tenover FC, White TJ, eds (1993): “Diagnostic Molecular Microbiology: Principles and Applications.” Washington, D.C.: American society for Microbiology. [Google Scholar]
  19. Sambrook J, Fritsch EF, Maniatis T (1989): “Molecular Cloning: A Laboratory Manual.” New York: Cold Spring Harbor Laboratory Press. [Google Scholar]
  20. Stanway G, Hughes PJ, Mountford RC, Minor PD, Almond JW (1984): The complete nucleotide sequence of a common cold virus: Human rhinovirus 14. Nucleic Acids Research 12: 7859–7875. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Medical Virology are provided here courtesy of Wiley

RESOURCES