title: Comparison of the conventional multiplex RT-PCR, real time RT-PCR and Luminex xTAG® RVP fast assay for the detection of respiratory viruses.
abstract: Detection of respiratory viruses using polymerase chain reaction (PCR) is sensitive, specific and cost effective, having huge potential for patient management. In this study, the performance of an in-house developed conventional multiplex RT-PCR (mRT-PCR), real time RT-PCR (rtRT-PCR) and Luminex xTAG(®) RVP fast assay (Luminex Diagnostics, Toronto, Canada) for the detection of respiratory viruses was compared. A total 310 respiratory clinical specimens predominantly from pediatric patients, referred for diagnosis of influenza A/H1N1pdm09 from August 2009 to March 2011 were tested to determine performance characteristic of the three methods. A total 193 (62.2%) samples were detected positive for one or more viruses by mRT-PCR, 175 (56.4%) samples by real time monoplex RT-PCR, and 138 (44.5%) samples by xTAG(®) RVP fast assay. The overall sensitivity of mRT-PCR was 96.9% (95% CI: 93.5, 98.8), rtRT-PCR 87.9% (95% CI: 82.5, 92.1) and xTAG(®) RVP fast was 68.3% (95% CI: 61.4, 74.6). Rhinovirus was detected most commonly followed by respiratory syncytial virus group B and influenza A/H1N1pdm09. The monoplex real time RT-PCR and in-house developed mRT-PCR are more sensitive, specific and cost effective than the xTAG(®) RVP fast assay.
authors.affiliation: National Institute of Virology, Human Influenza Group, Pune, India.
authors.forename: Manohar L
authors.initials: ML
authors.lastname: Choudhary
authors.affiliation: National Institute of Virology, Human Influenza Group, Pune, India.
authors.forename: Siddharth P
authors.initials: SP
authors.lastname: Anand
authors.affiliation: National Institute of Virology, Human Influenza Group, Pune, India.
authors.forename: Shamal A
authors.initials: SA
authors.lastname: Tikhe
authors.affiliation: National Institute of Virology, Human Influenza Group, Pune, India.
authors.forename: Atul M
authors.initials: AM
authors.lastname: Walimbe
authors.affiliation: National Institute of Virology, Human Influenza Group, Pune, India.
authors.forename: Varsha A
authors.initials: VA
authors.lastname: Potdar
authors.affiliation: National Institute of Virology, Human Influenza Group, Pune, India.
authors.forename: Mandeep S
authors.initials: MS
authors.lastname: Chadha
authors.affiliation: National Institute of Virology, Human Influenza Group, Pune, India.
authors.forename: Akhilesh C
authors.initials: AC
authors.lastname: Mishra
mesh_terms: D000818:Animals; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000821:virology*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000175:diagnosis / Q000821:virology*; D014780:Viruses / Q000302:isolation & purification*
keywords: RT-PCR; RVP assay; respiratory viruses; xTAG®