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.

Clinical Evaluation of the New High-Throughput Luminex NxTAG Respiratory Pathogen Panel Assay for Multiplex Respiratory Pathogen Detection

“…The lower observed sensitivity for CoV HKU-1 and CoV OC43 may be due to the low positive sample number for this target and the fact that NxTAG-RPP detected two of the three positives. This performance represented a significant improvement from the previous Luminex xTAG RVPv2 assay (15,16,22). However, the assay was found to be oversensitive, with false-positive results noted in 12 of 20 targets (60.0%) (adenovirus, HCoV-229E, HCoV-OC43, HCoV-NL63, enterovirus/rhinovirus, influenza A/H3, hMPV, PIV2, PIV3, PIV4, RSV, and HBoV).…”

Targeted Sequencing of Respiratory Viruses in Clinical Specimens for Pathogen Identification and Genome-Wide Analysis

“…The method is expected to detect all commonly known respiratory viral pathogens that are diagnosed with molecular tests such as Luminex xTAG RVP assay, FilmArray Respiratory Panel (RP) tests, multiplex real-time PCR tests for respiratory viral infection with comparable sensitivity and accuracy [9][10][11]. The large capture panel and the scheme of genome-wide capture allow detection of most known respiratory viruses and viruses with high sequence divergences.…”

Partial comparison of the NxTAG Respiratory Pathogen Panel Assay with the Luminex xTAG Respiratory Panel Fast Assay V2 and singleplex real-time polymerase chain reaction for detection of respiratory pathogens

“…The Luminex Respiratory Virus Panel (RVP) Fast Assay was the first version of an assay with the ability to simultaneously identify targets of 19 different viral types and subtypes in respiratory secretions. Although some studies have reported that the RVP Fast Assay may provide results comparable or superior to those of culture and DFA for the diagnosis of respiratory viral infections (Mahony et al, 2007;Pabbaraju et al, 2011), the RVP Fast Assay has been criticized because direct comparisons with real-time PCR have shown that discordant results may occur for some viruses, and reduced sensitivity occurs mainly in samples with low viral loads (Choudhary et al, 2016;Esposito et al, 2016b;Gadsby et al, 2010). To overcome these limitations, a new assay, the Luminex RVP Fast Assay v2, has been developed.…”