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.

scholarly journals Adenovirus, parainfluenza virus and respiratory syncytial virus antibodies in the sera of Jamaicans

1972 ◽  
Vol 70 (3) ◽  
pp. 523-529 ◽  
Author(s):  
Roy Jennings
Keyword(s):  
Respiratory Syncytial Virus ◽  
Respiratory Virus ◽  
Respiratory Viruses ◽  
High Titre ◽  
Parainfluenza Virus ◽  
Age Group ◽  
Syncytial Virus ◽  
Almost All ◽  
Type 3

SUMMARYSurveys for respiratory virus antibodies in the Jamaican population have shown that adenovirus, respiratory syncytial virus and parainfluenza types 1 and 3 virus antibodies are acquired early in life. The incidence of haemagglutination-inhibiting antibodies to parainfluonza viruses increases rapidly with age and almost all adults possess parainfluenza type 3 antibody, usually in high titre. Parainfluenza type 1 antibodies are only slightly less common. Complement-fixing antibodies to the adenovirus group were also observed to increase in incidence with age.Complement-fixing antibody to respiratory syncytial virus was less common in Jamaican sera than antibody to the other respiratory viruses described here. The highest titres were observed in the youngest age-group.

scholarly journals In-house standardization and validation of a multiplex RT-PCR assay for the detection of 13 respiratory viruses

Nova ◽  
2016 ◽  
Vol 14 (26) ◽  
pp. 9
Author(s):  
Hernán Vargas ◽  
Ángela Diaz ◽  
Yamile Celis ◽  
Liliana Díaz ◽  
Sandra Gómez ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Respiratory Viruses ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Lower Efficiency ◽  
Single Target ◽  
Syncytial Virus ◽  
Sensitivity Specificity

<p>Background. Multiplex real time PCR is increasingly used to diagnose respiratory viruses and has shown to be superior to traditional methods, such as culture and antigen detection. Objective. Standardization and validation of a multiplex real-time PCR assay for the detection of 13 respiratory viruses. Methods. The assay was validated using RNA control targets and comparing results to single-target PCR’s. Results. Using RNA controls the multiplex format was found to be as sensitive and specific as the single-target PCRs, and no competition was observed between targets. The efficiencies for most of the reactions were approximately 90%, but a lower efficiency was found for Parainfluenza 2 with a rate of amplification in each cycle of 86.63%. On the other hand, a higher efficiency was observed in respiratory syncytial virus A and respiratory syncytial virus B ((93.07% each). Conclusion: This multiplex RT-PCR format shows an adequate efficiency, demonstrating an excellent sensitivity, specificity and repeatability for all the studied respiratory viruses.</p>

scholarly journals Multicenter Clinical Evaluation of the Alere i Respiratory Syncytial Virus Isothermal Nucleic Acid Amplification Assay

2017 ◽  
Vol 56 (3) ◽  
Author(s):  
Ferdaus Hassan ◽  
Lindsay M. Hays ◽  
Aleta Bonner ◽  
Bradley J. Bradford ◽  
Ruffin Franklin ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Nucleic Acid ◽  
Care Setting ◽  
Point Of Care ◽  
Nucleic Acid Amplification ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Isothermal Nucleic Acid Amplification ◽  
Syncytial Virus ◽  
Respiratory Specimens

ABSTRACTThe Alere i respiratory syncytial virus (RSV) assay is an isothermal nucleic acid amplification test capable of detecting RSV directly from respiratory specimens, with results being available in ≤13 min after test initiation. The objective of this study was to evaluate the performance characteristics of the Alere i RSV assay in a point-of-care setting by using direct nasopharyngeal (NP) swab specimens (direct NP) and nasopharyngeal swab specimens eluted and transported in viral transport medium (VTM NP). The study was a prospective, multicenter, clinical trial conducted at 9 sites across the United States to evaluate the clinical performance of the Alere i RSV assay with respiratory specimens obtained from both children (age, <18 years) and older adults (age, >60 years). The performance of the Alere i RSV assay was compared with that of the reference method, the Prodesse ProFlu+ real-time reverse transcriptase PCR (RT-PCR) assay. All specimens with discrepant test results were tested further by a second FDA-cleared PCR assay (the Verigene respiratory virus plus nucleic acid test; Luminex Inc., TX). A total of 554 subjects with signs and symptoms of respiratory infections were enrolled, and respiratory samples were collected in this study. In comparison with the ProFlu+ real-time RT-PCR, the overall sensitivity and specificity of Alere i RSV assay for the detection of RSV were 98.6% (95% confidence interval [CI], 94.4 to 99.7%) and 98.0% (95% CI, 95.8 to 99.1%), respectively, for direct NP and 98.6% (95% CI, 94.4 to 99.7%) and 97.8% (95% CI, 95.5 to 98.9%), respectively, for VTM NP. The Alere i RSV is a highly sensitive and specific molecular assay ideal for rapid RSV detection in patients in the point-of-care setting due to its minimal hands-on time and rapid result availability.

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.

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