Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Jan;34(1):140–143. doi: 10.1128/jcm.34.1.140-143.1996

Diagnosis of viral respiratory tract infections in children by using a reverse transcription-PCR panel.

L L Gilbert 1, A Dakhama 1, B M Bone 1, E E Thomas 1, R G Hegele 1
PMCID: PMC228747  PMID: 8748290

Abstract

Reverse transcription-PCR (RT-PCR) is a sensitive method for detection of RNA virus nucleic acid sequences in clinical respiratory specimens. Previous studies have focused on RT-PCR for a single virus, but this approach is limited by the inability to establish a specific etiology when the RT-PCR result is negative and by the inability to document simultaneous infections involving more than one virus. The purpose of this study was to apply a panel of RT-PCR protocols for respiratory syncytial virus, parainfluenza virus, and picornaviruses to respiratory specimens from 80 children suspected to have acute viral respiratory tract infections and to correlate RT-PCR results with viral culture results and clinical diagnosis. In comparison with viral culture, the RT-PCR panel had a sensitivity of over 94% and showed evidence of simultaneous infections in a significantly greater proportion of specimens (20.0% versus 3.8%; P < 0.002). For specimens in which no viruses were detected by culture, the proportion of specimens with positive picornavirus RT-PCR results was significantly greater than the proportion of specimens with positive respiratory syncytial virus or parainfluenza virus RT-PCR results (P < 0.001). There were no statistically significant associations between RT-PCR results and clinical diagnosis. In summary, the RT-PCR panel provides an improved approach to obtain new insights into acute viral respiratory tract infections in children.

Full Text

The Full Text of this article is available as a PDF (255.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. CHANOCK R. M., PARROTT R. H. ACUTE RESPIRATORY DISEASE IN INFANCY AND CHILDHOOD: PRESENT UNDERSTANDING AND PROSPECTS FOR PREVENTION. Pediatrics. 1965 Jul;36:21–39. [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Claas E. C., Sprenger M. J., Kleter G. E., van Beek R., Quint W. G., Masurel N. Type-specific identification of influenza viruses A, B and C by the polymerase chain reaction. J Virol Methods. 1992 Sep;39(1-2):1–13. doi: 10.1016/0166-0934(92)90120-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cubie H. A., Inglis J. M., Leslie E. E., Edmunds A. T., Totapally B. Detection of respiratory syncytial virus in acute bronchiolitis in infants. J Med Virol. 1992 Dec;38(4):283–287. doi: 10.1002/jmv.1890380410. [DOI] [PubMed] [Google Scholar]
  5. Frick O. L., German D. F., Mills J. Development of allergy in children. I. Association with virus infections. J Allergy Clin Immunol. 1979 Apr;63(4):228–241. doi: 10.1016/0091-6749(79)90106-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hegele R. G., Hayashi S., Bramley A. M., Hogg J. C. Persistence of respiratory syncytial virus genome and protein after acute bronchiolitis in guinea pigs. Chest. 1994 Jun;105(6):1848–1854. doi: 10.1378/chest.105.6.1848. [DOI] [PubMed] [Google Scholar]
  7. Johnston S. L., Sanderson G., Pattemore P. K., Smith S., Bardin P. G., Bruce C. B., Lambden P. R., Tyrrell D. A., Holgate S. T. Use of polymerase chain reaction for diagnosis of picornavirus infection in subjects with and without respiratory symptoms. J Clin Microbiol. 1993 Jan;31(1):111–117. doi: 10.1128/jcm.31.1.111-117.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Karron R. A., Froehlich J. L., Bobo L., Belshe R. B., Yolken R. H. Rapid detection of parainfluenza virus type 3 RNA in respiratory specimens: use of reverse transcription-PCR-enzyme immunoassay. J Clin Microbiol. 1994 Feb;32(2):484–488. doi: 10.1128/jcm.32.2.484-488.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Karron R. A., O'Brien K. L., Froehlich J. L., Brown V. A. Molecular epidemiology of a parainfluenza type 3 virus outbreak on a pediatric ward. J Infect Dis. 1993 Jun;167(6):1441–1445. doi: 10.1093/infdis/167.6.1441. [DOI] [PubMed] [Google Scholar]
  10. McIntosh K., Ellis E. F., Hoffman L. S., Lybass T. G., Eller J. J., Fulginiti V. A. The association of viral and bacterial respiratory infections with exacerbations of wheezing in young asthmatic children. J Pediatr. 1973 Apr;82(4):578–590. doi: 10.1016/S0022-3476(73)80582-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ohlin A., Hoover-Litty H., Sanderson G., Paessens A., Johnston S. L., Holgate S. T., Huguenel E., Greve J. M. Spectrum of activity of soluble intercellular adhesion molecule-1 against rhinovirus reference strains and field isolates. Antimicrob Agents Chemother. 1994 Jun;38(6):1413–1415. doi: 10.1128/aac.38.6.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pattemore P. K., Johnston S. L., Bardin P. G. Viruses as precipitants of asthma symptoms. I. Epidemiology. Clin Exp Allergy. 1992 Mar;22(3):325–336. doi: 10.1111/j.1365-2222.1992.tb03094.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Subbarao E. K., Griffis J., Waner J. L. Detection of multiple viral agents in nasopharyngeal specimens yielding respiratory syncytial virus (RSV). An assessment of diagnostic strategy and clinical significance. Diagn Microbiol Infect Dis. 1989 Jul-Aug;12(4):327–332. doi: 10.1016/0732-8893(89)90098-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES