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. 2002 Nov 12;30(1):115–125. doi: 10.1016/0166-0934(90)90049-L

Detection of rhinovirus RNA in nasal epithelial cells by in situ hybridization

Christine Bruce 1, Patricia Chadwick 1, Widad Al-Nakib 1,2,∗
PMCID: PMC7119484  PMID: 1964938

Abstract

This paper describes the development and evaluation of in situ hybridization (ISH) for the detection of rhinovirus in cells obtained from nasal washings of volunteers infected with human rhinovirus 14 (HRV-14).

Twenty-five (66%) and 27 (71%) of 38 volunteers inoculated with HRV-14 had evidence of infection by virus isolation and ISH, respectively, on at least one of 4 days investigated after virus challenge. In contrast, only 14 of 38 (37%) volunteers had significant antibody rises as detected by the neutralization test.

Of the 38 volunteers inoculated with HRV-14, only 13 (34%) had symptoms of colds. Of these, 12 (92%) and 10 (77%) were positive by virus isolation or ISH, respectively, on at least one day. Six (46%) had significant antibody rises by neutralization. Similarly, of the 38 volunteers challenged, 22 (58%) were asymptomatic and of these 10 (45.5%) and 12 (54.5%) were positive by virus isolation and ISH, respectively, on at least one day. Only 8 (36.4%) of these asymptomatic volunteers showed significant antibody rises by neutralization.

There were significant associations between the detection of rhinoviruses by ISH and virus isolation on the third day (P<0.025) after virus challenge in the group as a whole and in the symptomatic group.

These results show that generally rhinovirus detection by ISH compares well with virus isolation and both tests are clearly more sensitive than the neutralization test in detecting evidence of infection. It is concluded that ISH is an interesting new technique that may play an important role in the study of rhinovirus infection and pathogenesis.

Keywords: Rhinoviruses, cDNA, Probes, Picornaviruses, In situ hybridization, Respiratory viral infections

References

  1. Al-Nakib W., Stanway G., Forsyth M., Hughes P.J., Almond J.W., Tyrrell D.A.J. Detection of human rhinoviruses and their molecular relationship using cDNA probes. J. Med. Virol. 1986;20:289–296. doi: 10.1002/jmv.1890200311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Al-Nakib W., Stanway G., Forsyth M., Hughes P.J., Almond J.W., Tyrrell D.A.J. Rhinovirus detection by cDNA:RNA hybridization. In: Brinton M.A., Rueckert R.R., editors. Positive Strand RNA Viruses. Vol. 54. Alan R. Liss. Inc; New York: 1987. pp. 487–495. (UCLA Symposia on Molecular and Cellular Biology, New Series). [Google Scholar]
  3. Al-Hakib W., Tyrrell D.A.J. Common cold viruses — Rhinoviruses. In: Ballows A., Hausler W.J., Lennette E.H., editors. Laboratory Diagnosis of Infectious Diseases: Principles and Practice. Springer-Verlag; New York: 1988. pp. 723–742. [Google Scholar]
  4. Beare A.S., Reed S.E. The study of antiviral compounds in volunteers. In: Oxford J.S., editor. Chemoprophylaxis and Virus Infections of the Respiratory tract. CRC Press; Cleveland: 1977. pp. 27–55. [Google Scholar]
  5. Bruce C.B., Al-Nakib W., Almond J.W., Tyrrell D.A.J. Use of synthetic oligonucleotide probes to detect rhinovirus RNA. Arch. Virol. 1989;105:179–187. doi: 10.1007/BF01311355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bruce C.B., Al-Nakib W., Forsyth M., Stanway G., Almond J.W. Detection of enteroviruses using cDNA and synthetic oligonucleotide probes. J. Virol. Methods. 1989;25:233–240. doi: 10.1016/0166-0934(89)90035-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burns J., Graham A.K., Frank C., Fleming K.A., Evans M.F., McGee J.O'D. Detection of low copy human papilloma virus DNA and mRNA in routine paraffin sections of cervix by non-isotopic in situ hybridization. J. Clin. Pathol. 1987;4:858–864. doi: 10.1136/jcp.40.8.858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Couch R.B. The common cold: control? J. Infect. Dis. 1984;150:167–173. doi: 10.1093/infdis/150.2.167. [DOI] [PubMed] [Google Scholar]
  9. Forghani B., Dupuis K.W., Schmidt N.J. Rapid detection of herpes simplex virus DNA in human brain tissue by in situ hybridization. J. Clin. Microbiol. 1985;22:656–658. doi: 10.1128/jcm.22.4.656-658.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Forsyth M., Al-Nakib W., Chadwick P., Stanway G., Hughes P.J., Leckie G., Almond J.W., Tyrrell D.A.J. Rhinovirus detection using probes from the 5' and 3' end of the genome. Arch. Virol. 1989;107:55–63. doi: 10.1007/BF01313878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gomes S.A., Nascimento J.P., Siqueira M.M., Krawczuk M.M., Pereira H.G., Russell W.C. In situ hybridization with biotinylated DNA probes: a rapid diagnostic test for adenovirus upper respiratory infections. J. Virol. Methods. 1985;12:105–110. doi: 10.1016/0166-0934(85)90012-6. [DOI] [PubMed] [Google Scholar]
  12. Gregg I. Provocation of airflow limitation by viral infection: implication for treatment. Eur. J. Resp. Dis. 1983;64:369–379. [PubMed] [Google Scholar]
  13. Jiang X.I., Estes M.K., Metcal F.G. In situ hybridization for quantitative assay of infectious hepatitis A virus. J. Clin. Microbiol. 1989;27:874–879. doi: 10.1128/jcm.27.5.874-879.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kandolf R., Ameis D., Krischner P., Canu A., Hofschneider P.H. Vol. 84. 1987. In situ detection of enteroviral genomes in myocardial cells by nucleic acid hybridization: an approach to the diagnosis of viral heart disease; pp. 6272–6276. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Krilov L., Pierik L., Keller E., Mahan K., Watson D., Hirsch M., Hamparian V., Mckintosh K. The association of rhinoviruses with lower respiratory tract disease in hospitalized patients. J. Med. Virol. 1986;19:345–352. doi: 10.1002/jmv.1890190407. [DOI] [PubMed] [Google Scholar]
  16. Myint S., Sidell S., Tyrrell D. Detection of Human Coronavirus 229E in nasal washing using RNA:RNA hybridization. J. Med. Virol. 1989;29:70–73. doi: 10.1002/jmv.1890290113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Naoumov N.V., Alexander G.J.M., O'Grady J.G., Aldis P., Portmann B.C., Williams R. Rapid diagnosis of cytomegalovirus infection by in situ hybridization in liver grafts. Lancet. 1988:1361–1364. doi: 10.1016/s0140-6736(88)92180-0. [DOI] [PubMed] [Google Scholar]
  18. Phillpotts R.J. Clones of MRC-C cells may be superior to the parent line for the culture of 229E-like strains of human respiratory coronavirus. J. Virol. Methods. 1983;6:267–269. doi: 10.1016/0166-0934(83)90041-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sperber S.J., Hayden F.G. Chemotherapy of Rhinovirus colds. Antimicrob. Agents Chemother. 1988;32:409–419. doi: 10.1128/aac.32.4.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tyrrell D.A.J., Barrow I., Arthur J. Local hyperthermia benefits natural and experimental common colds. Br. Med. J. 1989;298:1280–1283. doi: 10.1136/bmj.298.6683.1280. [DOI] [PMC free article] [PubMed] [Google Scholar]

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