Comparison of fast-track diagnostics respiratory pathogens multiplex real-time RT-PCR assay with in-house singleplex assays for comprehensive detection of human respiratory viruses

► FTDRP is a commercial multiplex real-time RT-PCR assay for 16 respiratory viruses. ► The FTDRP assay was compared with in-house singleplex real-time RT-PCR assays. ► Overall, the FTDRP and in-house assays performed comparably, with some exceptions. Fast-track Diagnostics respiratory pathogens (FTD...

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Bibliographic Details
Published in:Journal of virological methods 2012-11, Vol.185 (2), p.259-266
Main Authors: Sakthivel, Senthilkumar K., Whitaker, Brett, Lu, Xiaoyan, Oliveira, Danielle B.L., Stockman, Lauren J., Kamili, Shifaq, Oberste, M. Steven, Erdman, Dean D.
Format: Article
Language:English
Subjects:
Adenoviridae - genetics
Adenoviridae - isolation & purification
Adenovirus
Adenovirus Infections, Human - diagnosis
Adenovirus Infections, Human - virology
Biological and medical sciences
Children
diagnostic techniques
Enterovirus
Fundamental and applied biological sciences. Psychology
Humans
instrumentation
Integration
Microbiology
Multiplex
Nasopharyngeal Diseases - diagnosis
Nasopharyngeal Diseases - virology
Pathogens
Picornaviridae Infections - diagnosis
Picornaviridae Infections - virology
Polymerase chain reaction
Prospective Studies
Quality control
quantitative polymerase chain reaction
Reagent Kits, Diagnostic
Real-time PCR
Real-Time Polymerase Chain Reaction - methods
Respiratory syncytial virus
Respiratory Syncytial Virus Infections - diagnosis
Respiratory Syncytial Virus Infections - virology
Respiratory Syncytial Virus, Human - genetics
Respiratory Syncytial Virus, Human - isolation & purification
Respiratory virus
reverse transcriptase polymerase chain reaction
Reverse Transcriptase Polymerase Chain Reaction - methods
Rhinovirus
Rhinovirus - genetics
Rhinovirus - isolation & purification
Sensitivity and Specificity
Techniques used in virology
Time Factors
Viral Load
Virology
viruses
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Summary:► FTDRP is a commercial multiplex real-time RT-PCR assay for 16 respiratory viruses. ► The FTDRP assay was compared with in-house singleplex real-time RT-PCR assays. ► Overall, the FTDRP and in-house assays performed comparably, with some exceptions. Fast-track Diagnostics respiratory pathogens (FTDRP) multiplex real-time RT-PCR assay was compared with in-house singleplex real-time RT-PCR assays for detection of 16 common respiratory viruses. The FTDRP assay correctly identified 26 diverse respiratory virus strains, 35 of 41 (85%) external quality assessment samples spiked with cultured virus and 232 of 263 (88%) archived respiratory specimens that tested positive for respiratory viruses by in-house assays. Of 308 prospectively tested respiratory specimens selected from children hospitalized with acute respiratory illness, 270 (87.7%) and 265 (86%) were positive by FTDRP and in-house assays for one or more viruses, respectively, with combined test results showing good concordance (K=0.812, 95% CI=0.786–0.838). Individual FTDRP assays for adenovirus, respiratory syncytial virus and rhinovirus showed the lowest comparative sensitivities with in-house assays, with most discrepancies occurring with specimens containing low virus loads and failed to detect some rhinovirus strains, even when abundant. The FTDRP enterovirus and human bocavirus assays appeared to be more sensitive than the in-house assays with some specimens. With the exceptions noted above, most FTDRP assays performed comparably with in-house assays for most viruses while offering enhanced throughput and easy integration by laboratories using conventional real-time PCR instrumentation.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2012.07.010