Diagnostic Accuracy of Seven Commercial Assays for Rapid Detection of Group A Rotavirus Antigens

Abstract

Seven commercial immunochromatographic assays intended for the detection of group A rotavirus antigens in human stool samples were evaluated. These assays showed similar levels of diagnostic accuracy and were suitable for the detection of rotavirus in patients with acute gastroenteritis but missed some asymptomatic rotavirus shedding identified by real-time reverse transcription-PCR. Group A rotaviruses (RVA) are a leading cause of severe gastroenteritis in young children worldwide, with an estimated 450,000 deaths of children under 5 years of age per year (1). Immunochromatographic (ICT) assays are widely used to detect RVA antigens in stool samples. A recent Australian study (2) demonstrated unexpectedly low specificity of the Vikia Rota-Adeno ICT assay (bioMérieux, Marcy l'Etoile, France), while excellent specificity was reported for this assay in previous studies (3,4). This observation underlines the importance of regular assessments of the diagnostic performance of ICT assays in order to accurately diagnose RVA infections. In this study, we compared the diagnostic accuracies of seven commercially available ICT assays intended for the detection of RVA antigens in human stool samples, including the Vikia Rota-Adeno ICT assay.The diagnostic accuracy of ICT assays was first assessed on a collection of surplus material from raw fecal samples collected for RVA screening by three French hospital laboratories (CharlevilleMézières, Dijon, and Saint-Etienne) during the RVA epidemic peak from February through May 2014. Whatever the rotavirus screening result, only specimens with enough material for the seven ICT assays and the reference test were included. Of note, because the asymptomatic individuals were not excluded, this first collection included patients with or without acute gastroenteritis (AGE) symptoms. Specimens were immediately stored at Ϫ20°C. The ICT assays were simultaneously performed on thawed samples from August to October 2014 at the National Reference Centre for Enteric Viruses, University Hospital of Dijon, France (NRCev), according to the manufacturer's instructions. The results of the assays were read by two laboratory technicians (plus a third laboratory technician in cases of discrepancies) blinded to the results of the initial rotavirus screening. The reference test was performed on the same thawed samples and on the same day as the ICT assays. This was an in-house real-time reverse transcriptionquantitative PCR (RT-qPCR) assay adapted from the literature (5) and allowed the simultaneous detection of the RVA VP2 coding gene and pAW109 inhibition control RNA (Applied Biosystems, Foster City, CA, USA). The rotavirus strains were G and P genotyped according to the EuroRotaNet methods (www .eurorota.net/docs.php). For each ICT assay, the sensitivity, specificity, likelihood ratios, and diagnostic odds ratio were calculated using Meta-DiSc software (6) and the statistical analysis (KruskalWallis nonparametric test) was performed with Stata software (StataCorp release 12, College Station, TX, USA)....

Diagnostic accuracy of VIKIA® Rota-Adeno and Premier™ Rotaclone® tests for the detection of rotavirus in Niger

“…The sensitivity of both assays was in accordance with the results of previous studies using RT-PCR as reference standard [ 7 – 9 ]. This reduced sensitivity compared to what has been reported in the manufacturer inserts for both assays could be due to the lack of detection of certain genotypes by EIA and RDT.…”

“…However, the 23 false negative specimens by both EIA and RDT were genotyped and showed the most frequent genotypes in our study (10 G2P[4], 8 G12P[8], 2 G3P[6], 2 G3+G9P[6] and 1 G9P[8]) [ 3 ], suggesting no clear association between lack of detection and particular genotypes. Instead, it is most probably due to the high sensitivity of the reference standard [ 8 , 9 ] and notably to the ability of RT-PCR to detect samples with low viral loads that cannot be detected by RDT and EIA assays [ 7 , 12 ]. Unfortunately, the rotavirus viral load in samples was not assessed during this study to confirm this hypothesis.…”

“…Initial evaluations of rotavirus detection by EIA and RDT mainly used culture, electron microscopy or viral RNA electrophoresis as the gold standard with good sensitivity and specificity [ 5 , 6 ]. Recent evaluations using highly sensitive reference methods as reverse transcription polymerase chain reaction (RT-PCR) have shown lower sensitivity of RDTs and EIAs for the detection of group A rotavirus in stool specimens [ 7 , 8 ]. Few studies have evaluated RDT and EIA tests in parallel to compare their performance on the same samples and showed similar performance for both tests [ 8 , 9 ].…”

“…Few studies have evaluated RDT and EIA tests in parallel to compare their performance on the same samples and showed similar performance for both tests [ 8 , 9 ]. While some studies reported a high level of false positive results with the widely used RDT VIKIA® Rota-Adeno (bioMérieux, Marcy l’Etoile, France) [ 8 , 10 ], other studies showed good sensitivity and specificity [ 7 , 11 ].…”

Prevalence and Genetic Diversity of Enteric Viruses in Children with Diarrhea in Ouagadougou, Burkina Faso

“…This could be explained by the detection method used in the present study. Indeed, real-time RT-PCR has been demonstrated to be more sensitive than antigen detection by immunochromatographic test [ 41 ]. The circulating G and P genotypes showed a diverse pattern, with G9P[ 8 ] strains dominating.…”