Diagnostic Accuracy and Analytical Sensitivity of IDEIA Norovirus Assay for Routine Screening of Human Norovirus

Abstract

Noroviruses (NoVs) are recognized as the leading cause of epidemic and sporadic acute gastroenteritis. Early detection of NoV is crucial to control the spread of the disease. In this study, we evaluated the diagnostic accuracy, analytical sensitivity, and analytical reactivity of the IDEIA Norovirus assay (an enzyme immunoassay [EIA]) in a prospective and retrospective study design. A total of 557 prospectively collected fecal samples and a panel of 97 archived fecal samples, including 21 different GI and GII genotypes, were tested by conventional reverse transcription-PCR (RT-PCR)/bidirectional sequencing, real-time RT-PCR, and electron microscopy. The sensitivity and specificity of the EIA were 57.6% and 91.9%, respectively. The sensitivity for detecting NoV in fecal samples from outbreaks improved from 44.1% when three samples were tested to 76.9% when five samples per outbreak were tested. The EIA was able to detect strains from 7 GI and 11 GII genotypes. The analytical sensitivity of the EIA was 3.1 ؋ 10 6 and 1.6 ؋ 10 7 virus particles g ؊1 of fecal sample for NoV GI and GII strains, respectively. Most GII samples positive by EIA had a threshold cycle (C T ) of <26.5, and 50% of the GII samples negative by EIA had a C T of >25.6, suggesting that, although strains from genotypes GI.8, GII.10, and GII.16 were not detected, the low sensitivity of the EIA is primarily caused by low virus concentration. In conclusion, the current EIA may be of use as a rapid screening test during a norovirus outbreak investigation when multiple fecal samples are available; however, sporadic samples should be tested by molecular methods.

Performance characteristics of enzyme linked immunosorbent assay and rapid immunochromatographic test for routine screening of human norovirus

“…They found a sensitivity 90.4%, specificity 96.4%, PPV 88.0%, NPV 97.2% and accuracy 95.0%. Costantini et al (6) found that the sensitivity and specificity of the IDEIA Norovirus kit ranged from 57.4% to 77.8% and 79.7% to 91.9%, respectively, when the results were compared by different reference standards.…”

“…Norwalk virus was discovered in 1972 by electron microscopy, then cloning and sequencing in 1990 occur. Diagnosis of NoV was done by conventional reverse RT-PCR assay in the mid-1990s followed by real-time RT-PCR (rRT-PCR) assays, which is now the method used for diagnosis of NoV in most clinical laboratories (6).The role of NoV as an important cause of both epidemic and sporadic gastroenteritis were better understood using molecular techniques and sequence analysis (7). Wide differences within this genus were detected using genomic analysis.…”

“…These kits require antibodies that cross react between NoV genogroups to overcome the high antigenic diversity of NoVs and help to detect new strains. The usefulness of these kits for routine screening of samples should be investigated because the sensitivity and specificity of NoV diagnostic kits vary with the diagnostic goal (outbreak or sporadic cases) and test design (6).…”

Rational Design of Duplex Specific Nuclease for One-Step Isothermal Viral RNA Detection

“…However, given the limited laboratory options for propagation of norovirus [55,56] and commercial kits for rapid diagnosis remain underdeveloped [57,58]. As a result, clinical laboratories commonly use reverse transcription-polymerase chain reaction (RT-PCR), immunoassays or hybridization assays [59][60][61][62], which can lead to false positives, provide lower sensitivities and require significant time and materials costs [62][63][64][65][66][67][68][69][70].…”

Design and Evaluation of Three Immuno-based Assays for Rapid Detection of Human Norovirus Virus-like Particles

“…ELISAs can analyze a myriad of pathological samples using multiple screening modes and reporter molecules, giving the assay broad versatility, but their need for high viral loads [5,27,30] limits the assays' application mostly in clinical settings. Despite their relatively lower sensitivity when compared to PCR-based methods [5,20], ELISA-based assays have been used to detect norovirus in human stool samples [5,25,[36][37][38][39][40][41][42][43], human sera [29], and food samples [26,31,32,44,45] Using norovirus GI.1 (Norwalk) virus-like particles (VLPs) as a model viral system, the objective of this study was to develop and evaluate several ELISA-based assays for rapid detection of varying concentrations of GI.1 VLPs (0.037-3.7 μg/mL). HuNoV VLPs are replication-incompetent, macromolecular protein assemblies with capsid structures and antigenic properties resembling those of innate norovirus particles [46][47][48].…”