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Journal of Clinical Microbiology, June 1999, p. 1858-1862, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Evaluation of a Nested Reverse Transcription-PCR Assay Based on the Nucleoprotein Gene for Diagnosis of Spontaneous and Experimental Bovine Respiratory Syncytial Virus Infections

Jean-François Valarcher,1 Hervé Bourhy,2 Jacqueline Gelfi,3 and François Schelcher1,*

UMR Institut National de la Recherche Agronomique-Ecole Nationale Vétérinaire de Toulouse de Physiopathologie Infectieuse et Parasitaire des Ruminants1 and UMR Institut National de la Recherche Agronomique-Ecole Nationale Vétérinaire de Toulouse de Microbiologie Moléculaire,3 ENVT, 31076 Toulouse cedex 3, and Unité de la Rage, Institut Pasteur, 75724 Paris cedex 15,2 France

Received 17 November 1998/Returned for modification 8 February 1999/Accepted 1 March 1999

The first nested reverse transcription (RT)-PCR based on the nucleoprotein gene (n RT-PCR-N) of the bovine respiratory syncytial virus (BRSV) has been developed and optimized for the detection of BRSV in bronchoalveolar lavage fluid cells of calves. This test is characterized by a low threshold of detection (0.17 PFU/ml), which is 506 times lower than that obtained by an enzyme immunosorbent assay (EIA) test (RSV TESTPACK ABBOTT). During an experimental infection of 17 immunocompetent calves less than 3 months old, BRSV RNA could be detected up to 13 days after the onset of symptoms whereas isolation in cell culture was possible only up to 5 days. Compiling results obtained by conventional techniques (serology, antigen detection, and culture isolation) for 132 field samples collected from calves with acute respiratory signs revealed that n RT-PCR-N showed the highest diagnostic sensitivity and very good specificity. This n RT-PCR-N with its long period of detection during BRSV infection thus provides a valuable tool for diagnostic and epidemiological purposes.

* Corresponding author. Mailing address: UMR INRA-ENVT de Physiopathologie Infectieuse et Parasitaire des Ruminants, ENVT, 23 Chemin des Capelles, 31076 Toulouse cedex 3, France. Phone: 33 05-61-19-38-33. Fax: 33 05-61-19-38-34. E-mail: f.schelcher{at}envt.fr.

Journal of Clinical Microbiology, June 1999, p. 1858-1862, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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