+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Evaluation of a commercial real-time reverse transcription polymerase chain reaction kit for the diagnosis of Bovine respiratory syncytial virus infection



Evaluation of a commercial real-time reverse transcription polymerase chain reaction kit for the diagnosis of Bovine respiratory syncytial virus infection



Journal of Veterinary Diagnostic Investigation 22(2): 238-241



Recently a commercial real-time reverse transcription polymerase chain reaction (RT-PCR) kit has been marketed for the detection of Bovine respiratory syncytial virus (BRSV). However, diagnostic interpretation of the results of this kit requires its comparison to commonly used methods. Therefore, the objective of this study was to evaluate the performance of this kit in comparison with the conventional direct fluorescent antibody test (FAT). Twenty BRSV strains and 14 heterologous bovine viruses were used to check the kit's sensitivity and specificity. The efficiency and detection limit of the kit were determined by testing dilution series of a BRSV strain. The comparison between real-time RT-PCR kit and FAT was performed with 94 clinical samples from calves with clinical signs of respiratory disease including lung tissues (n = 55), transtracheal aspiration samples (n = 20), and nasal swab samples (n = 19). All of the BRSV strains tested were detected by real-time RT-PCR. No cross-reaction was shown with the 14 heterologous bovine viruses. The real-time RT-PCR was 99.3% efficient with a detection limit of 0.1 TCID(50) (50% tissue culture infective dose). The results of real-time RT-PCR and FAT were concordant for 65 of the 94 clinical samples tested. The remaining 29 clinical samples were positive by real-time RT-PCR and negative by FAT, demonstrating the higher sensitivity of real-time RT-PCR. In conclusion, the kit evaluated in this study was sensitive, specific, and had a low threshold of detection. Furthermore, the use of this kit instead of FAT allows an improvement of the sensitivity for the detection of BRSV in clinical samples.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 053062868

Download citation: RISBibTeXText

PMID: 20224083

DOI: 10.1177/104063871002200211


Related references

Evaluation of consistency in quantification of gene copy number by real-time reverse transcription quantitative polymerase chain reaction and virus titer by plaque-forming assay for human respiratory syncytial virus. Microbiology and Immunology 62(2): 90-98, 2018

Development and evaluation of a quantitative real time reverse transcriptase polymerase chain reaction assay for respiratory syncytial virus. Abstracts of the Interscience Conference on Antimicrobial Agents & Chemotherapy 43: 498, 2003

Reverse transcription polymerase chain reaction (RT-PCR) for diagnosis of respiratory syncytial virus infection in adults: use of a single-tube hanging droplet nested PCR. Journal of Medical Virology 63(3): 259-263, 2001

Utility of real time polymerase chain reaction in the diagnosis of respiratory syncytial virus infection among adult patients. Revista Chilena de Infectologia 24(6): 441-445, 2007

Development of a real time reverse transcriptase polymerase chain reaction for the detection of bovine respiratory syncytial virus in clinical samples and its comparison with immunohistochemistry and immunofluorescence antibody testing. Veterinary Microbiology 126(1-3): 264-270, 2008

Identifying bovine respiratory syncytial virus by reverse transcription-polymerase chain reaction and oligonucleotide hybridizations. Journal of Clinical Microbiology 31(5): 1237-1240, 1993

Diagnosis of Enzootic Pneumonia in Danish Cattle: Reverse Transcription-Polymerase Chain Reaction Assay for Detection of Bovine Respiratory Syncytial Virus in Naturally and Experimentally Infected Cattle. Journal of Veterinary Diagnostic Investigation 11(5): 416-422, 1999

Diagnosis of enzootic pneumonia in Danish cattle: reverse transcription-polymerase chain reaction assay for detection of bovine respiratory syncytial virus in naturally and experimentally infected cattle. Journal of Veterinary Diagnostic Investigation 11(5): 416-422, 1999

Optimization of the elution buffer and concentration method for detecting hepatitis E virus in swine liver using a nested reverse transcription-polymerase chain reaction and real-time reverse transcription-polymerase chain reaction. Journal of Virological Methods 206: 99, 2014

A new high-speed droplet-real-time polymerase chain reaction method can detect bovine respiratory syncytial virus in less than 10 min. Journal of Veterinary Medical Science 76(3): 477-480, 2014

Comparison of real-time reverse transcription loop-mediated isothermal amplification and real-time reverse transcription polymerase chain reaction for duck Tembusu virus. Journal of Virological Methods 182(1-2): 50-55, 2012

Comparison of FilmArray Respiratory Panel and laboratory-developed real-time reverse transcription-polymerase chain reaction assays for respiratory virus detection. Diagnostic Microbiology and Infectious Disease 74(4): 379-383, 2012

Dual Combined Real-Time Reverse Transcription Polymerase Chain Reaction Assay for the Diagnosis of Lyssavirus Infection. Plos Neglected Tropical Diseases 10(7): E0004812, 2016

Respiratory syncytial virus outbreak in a long-term care facility detected using reverse transcriptase polymerase chain reaction: an argument for real-time detection methods. Journal of the American Geriatrics Society 57(3): 482-485, 2009

Comparison of real-time reverse transcriptase-polymerase chain reaction and nested or commercial reverse transcriptase-polymerase chain reaction for the detection of hepatitis E virus particle in human serum (vol 56, pg 269, 2006). Diagnostic Microbiology and Infectious Disease 57(3): 353, 2007