Diagnostic performance of two highly multiplexed respiratory virus assays in a pediatric cohort

Abstract

Background Rapid detection of respiratory viruses is important for management and infection control in hospitalized patients. Multiplex nucleic acid tests (NATs) have begun to replace conventional methods as gold standards for respiratory virus detection. Objective To compare the performance of two large multiplex NATS, ResPlex II (RPII) and Respiratory Virus Surveillance kit with electrospray ionization mass spectrometry (RVS/MS) using nasopharyngeal aspirates (NPAs) from hospitalized children who had been tested previously with conventional methods. Study design Stored residual NPAs (N = 306) were tested concomitantly by RPII and RVS/MS. Alternate NATs were used to adjudicate discordant results. Results More viruses were detected with multiplex NATs (RPII, 110; RVS/MS, 109) than conventional assays (86); diagnostic gain was primarily for fastidious viruses (coronaviruses and enteroviruses [EVs]/human rhinoviruses [HRVs]). Total positive and negative agreement between the multiplex NATs for all viruses detected was quite high (86% positive agreement, 99% negative agreement). Most individual viruses were detected with fairly equivalent accuracy by the multiplex NATs, except for adenoviruses (RPII sensitivity 40%) and human metapneumovirus (RVS/MS sensitivity 42%). RPII had the advantage of detecting EVs and HRVs, however, it demonstrated considerable EV/HRV cross-reactivity (29 HRV-positive specimens by real-time PCR were positive for EV by RPII and 21 specimens positive for HRV only by RT-PCR were dual positive for EV/HRV by RPII). RPII also had reduced sensitivity for HRV detection (in 36 specimens, HRV was detected by RT-PCR but not by RPII). Conclusions Both multiplex NATs were promising, but had notable limitations.

Distribution of respiratory viruses which cause lower respiratory tract infection in pediatric age group

“…Though the low sensitivities and specificities are reported between about 10%-75%, these multiplex RT-PCR kits have been widely used in diagnosis [2,3,5]. The low sensitivity rates of the kits have been frequently reported for H1N1-p, influenzae A, enterovirus, CoV (OC43), and RSV depending on the reference standards due to lack of careful optimization, low viral load, and the patient population of the study.…”

“…In addition, RT-PCR is easy to perform and require no experienced staff [3,5]. In the present study, it was aimed to determine the distribution of viral causes in children admitted to the Department of Pediatrics of Rize Training and Research Hospital between January 2014 and January 2015 with pre-diagnosis of RTI.…”

“…However, in recent years, development and common usage of new antiviral agents have couraged the determination of viruses in cases of RTIs. In addition, epidemiological researches increased the requirement of viral detection [2,3,5].…”

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“…24 In two other studies, the coinfection rates of HRV-EV were found to be higher, compared with other agents. 25,26 As seen in all these studies, why viruses lead to coinfections still remains unclear.…”

Evaluation of the Pan Influenza detection kit utilizing the PLEX-ID and influenza samples from the 2011 respiratory season

“…The PLEX-ID platform has shown previously to be a useful tool for respiratory virus surveillance and Influenza A virus detection, identification and genotyping (Chen et al, 2011a,b; Deyde et al, 2010; Forman et al, 2012; Tang et al, 2013; Jeng et al, 2012; Sampath et al, 2007a,b). Updated assay plates including hemagglutinin (HA) and neuraminidase (NA) primers were developed recently to comply with recommendations from regulatory agencies and to provide an immediate characterization of H1N1 pdm09.…”