scholarly journals Preliminary Evaluation of a Multiplex Reverse Transcription-PCR Assay Combined with a New DNA Chip Hybridization Assay for Detecting Respiratory Syncytial Virus

2007 ◽  
Vol 45 (11) ◽  
pp. 3811-3813 ◽  
Author(s):  
M. Causse ◽  
A. D. Garcia-Mayorgas ◽  
J. B. Gutierrez ◽  
M. Casal
Keyword(s):  
Respiratory Syncytial Virus ◽  
Reverse Transcription ◽  
Dna Chip ◽  
Hybridization Assay ◽  
Preliminary Evaluation ◽  
Pcr Assay ◽  
Reverse Transcription Pcr ◽  
Syncytial Virus

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

1999 ◽  
Vol 37 (6) ◽  
pp. 1858-1862 ◽  
Author(s):  
Jean-François Valarcher ◽  
Hervé Bourhy ◽  
Jacqueline Gelfi ◽  
François Schelcher
Keyword(s):  
Respiratory Syncytial Virus ◽  
Reverse Transcription ◽  
Lavage Fluid ◽  
Bovine Respiratory Syncytial Virus ◽  
Virus Infections ◽  
Rt Pcr ◽  
Nucleoprotein Gene ◽  
Reverse Transcription Pcr ◽  
Field Samples ◽  
Syncytial Virus

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.

scholarly journals Direct Detection of Respiratory Syncytial Virus, Parainfluenza Virus, and Adenovirus in Clinical Respiratory Specimens by a Multiplex Reverse Transcription-PCR Assay

1998 ◽  
Vol 36 (11) ◽  
pp. 3149-3154 ◽  
Author(s):  
Carla Osiowy
Keyword(s):  
Tissue Culture ◽  
Respiratory Syncytial Virus ◽  
Respiratory Virus ◽  
Respiratory Viruses ◽  
Parainfluenza Virus ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Reverse Transcription Pcr ◽  
Syncytial Virus ◽  
Respiratory Specimens

Diagnosis of respiratory virus infections currently involves detection by isolation or antigen detection, which usually identifies only a single suspected agent. To permit identification of more than one respiratory virus in clinical specimens, a rapid detection method involving a single-step, multiplex reverse transcription-PCR (RT-PCR) assay was developed. The assay included five primer sets that amplified the RNA of respiratory syncytial virus subtypes A and B, parainfluenza virus types 1, 2, and 3, and adenovirus types 1 to 7. Initially the assay was tested on tissue culture-grown virus and was found to be specific for all 12 prototype viruses tested, with no interassay cross amplification or amplification of other respiratory viruses. Assay sensitivity allowed a detection range of 0.2 50% tissue culture infectious dose (TCID50) for adenovirus to 250 TCID50 for parainfluenza virus type 1. The multiplex RT-PCR assay was also able to directly detect viruses in respiratory specimens, with virus being detected in 41 of 112 samples as compared to 34 of 112 samples detected by direct immunofluorescence or antigen detection following specimen culture. This suggests that the multiplex RT-PCR assay can be used as a rapid and sensitive diagnostic method for major respiratory viruses.

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