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Journal of Clinical Microbiology, September 2003, p. 4298-4303, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4298-4303.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

GeneScan Reverse Transcription-PCR Assay for Detection of Six Common Respiratory Viruses in Young Children Hospitalized with Acute Respiratory Illness

Dean D. Erdman,^1* Geoffrey A. Weinberg,² Kathryn M. Edwards,³ Frances J. Walker,¹ Barbara C. Anderson,¹ Jörn Winter,¹ Monica González,⁴ and Larry J. Anderson¹

Centers for Disease Control and Prevention, Atlanta, Georgia 30333,¹ Department of Pediatrics, University of Rochester, Rochester, New York 14642,² Pediatrics Department, Vanderbilt University, Nashville, Tennessee 37235,³ Hospital Central de las Fuerzas Armadas, Montevideo, Uruguay⁴

Received 1 May 2003/ Returned for modification 27 June 2003/ Accepted 1 July 2003

A reverse transcription-PCR (RT-PCR) assay based on automated fluorescent capillary electrophoresis and GeneScan software analysis was developed to detect six common respiratory viruses in clinical specimens from young children. Assays for human respiratory syncytial virus (HRSV); human parainfluenza viruses 1, 2, and 3 (HPIV1, -2, and -3, respectively); and influenza A and B viruses were incorporated into a single standard assay format. The optimized assay panel was used to test 470 respiratory specimens obtained from 462 children hospitalized with acute respiratory illness that had been previously tested by viral culture (405 specimens) or direct immunofluorescence staining (DIF) (65 specimens). Of 93 specimens positive for respiratory viruses by culture or DIF, 86 (92%) were positive by RT-PCR, including 66 HRSV, 2 HPIV2, 5 HPIV3, 3 influenza A virus, and 10 influenza B virus specimens. An additional 119 respiratory viruses were identified by RT-PCR in 116 patients for whom results were negative by viral isolation or DIF. We conclude that the GeneScan RT-PCR panel can markedly improve detection of acute respiratory virus infections in young children.

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* Corresponding author. Mailing address: Respiratory and Enteric Viruses Branch, Mailstop G-09, Centers for Disease Control and Prevention, 1600 Clifton Rd., N.E., Atlanta, GA 30333. Phone: (404) 639-3727. Fax: (404) 639-1307. E-mail: dde1{at}cdc.gov.

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Journal of Clinical Microbiology, September 2003, p. 4298-4303, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4298-4303.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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