![]() ![]() Received: FebruAccepted: NovemPublished: November 16, 2015Ĭopyright: © 2015 Parker et al. PLoS ONE 10(11):Įditor: Lars Kaderali, University Medicine Greifswald, GERMANY (2015) Analytical Sensitivity Comparison between Singleplex Real-Time PCR and a Multiplex PCR Platform for Detecting Respiratory Viruses. The relationship between TCID 50/mL concentrations and the corresponding copy number related to various ATCC cultures is also reported.Ĭitation: Parker J, Fowler N, Walmsley ML, Schmidt T, Scharrer J, Kowaleski J, et al. Broader detection of influenza A/H3 viruses was demonstrated by the GenMark eSensor RVP. The multiplex PCR platform, the GenMark eSensor RVP, demonstrated improved analytical sensitivity for detecting influenza A/H3 viruses, influenza B virus, human parainfluenza virus 2, and human rhinovirus (1.6–94.8 copies/μL, 0.20–1.98 logs). Largest differences in LOD were demonstrated for assays targeting adenovirus group E, respiratory syncytial virus subtype A, and a generic assay for all influenza A viruses regardless of subtype (319.4–1280.8 copies/μL, 2.50–3.11 log difference). Assays targeting influenza A/H3N2, influenza A/H1N1pdm09, influenza B, and human parainfluenza 1 and 2 were most comparable (1.2–8.4 copies/μL, <1 log difference). Analytical sensitivity between the two methods varied from 1.2–1280.8 copies/μL (0.08–3.11 log differences) for all 12 assays compared. The GenMark eSensor RVP LODs were obtained by converting the TCID 50/mL concentrations reported in the package insert to copies/μL using qPCR. LODs were measured for each singleplex real-time PCR assay and expressed as the lowest copy number detected 95–100% of the time, depending on the assay. In this experiment, the lower limit of detection (LOD) of singleplex real-time PCR assays targeting respiratory viruses is compared to an equivalent panel on a multiplex PCR platform, the GenMark eSensor RVP. ![]() However, multiplexed assays must be well optimized to retain or improve upon the analytic sensitivity of their singleplex counterparts. Multiplex PCR methods are attractive to clinical laboratories wanting to broaden their detection of respiratory viral pathogens in clinical specimens. ![]()
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