Evaluation of a commercial ELISA for detecting Norwalk-like virus antigen in faeces

Richards, Alison F.; Lopman, Ben; Gunn, Annabel; Curry, Alan; Ellis, D. A.; Cotterill, Hilary; Ratcliffe, SJ; Jenkins, Mary; Appleton, Hazel; Gallimore, Christopher; Gray, Jim; Brown, David

doi:10.1016/s1386-6532(02)00267-6

Cited by 96 publications

(66 citation statements)

References 19 publications

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“…13 In addition the gut flora in neonates and especially premature neonates could differ from infants and adults, thus explaining discrepancies between the DAKO kit's evaluation trials and our results, 14 but this explanation is unlikely given that this test is licensed for use in humans, including neonates. Based on the data described above, we question the high specificity of certain norovirus antigen EIA suggested in published investigations, 7,15 particularly considering the consequences, that is, the cost and logistic problems caused by the hygienic measures such as ward closures resulting from positive NoV results. We call for studies into the specificity of these tests for norovirus detection in stool samples from neonates.…”

Section: Discussionmentioning

confidence: 98%

Apparently non-specific results found using a norovirus antigen immunoassay for fecal specimens from neonates

et al. 2007

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Norovirus is increasingly recognized as a frequent cause of non-bacterial gastroenteritis. Despite a 10-fold increase in the number of cases reported following the availability of enzyme immunoassays, there are no reports yet from preterm neonates. We report on a sudden clustering of antigen-positive enzyme immuno assays results in a level III neonatal intensive care unit, involving 22 of 43 infants screened. Although antigen-positive samples were significantly associated with bloody stools (P<0.001) and gastric residues (P<0.02), norovirus infection could not be confirmed by reversetranscriptase polymerase chain reaction or electron microscopy. We question the validity of the so called norovirus-specific antigen assays and warn against overreacting to positive enzyme immunoassays results without reverse-transcriptase polymerase chain reaction confirmation especially in the neonatal setting.

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Section: Discussionmentioning

confidence: 98%

Apparently non-specific results found using a norovirus antigen immunoassay for fecal specimens from neonates

et al. 2007

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“…First, collection of samples after onset of symptoms by three days may play a role in lack of detection of NoV (7). Furthermore, long periods of fecal storage before testing may also affect the level of the virus, as proteolytic degradation may take place (24,25).Collection and examination of fecal samples within 72 h after onset of viral gastroenteritis symptoms was done in our study, while in other studies samples were tested after storage for 2 to 3 years (26). Second, the outcome of all studies may be affected by differences in inclusion criteria fecal samples.…”

Section: In This Workmentioning

confidence: 99%

“…Some studies included specimens only when the clinical symptoms indicated a NoV outbreak. On the other hand some studies tested all samples that were sent to the laboratory (24,25). Finally, the discrepancy in sensitivity of Rida quick and EIA kit found in different studies may be explained by the antigenic diversity of NoV.Certain genotypes may be missed becausethese kits use monoclonal and polyclonal antibodies formed against virus-like particles representing GI and GII genotypes.…”

Section: In This Workmentioning

confidence: 99%

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

Sharaf¹,

Morsi²,

Gerges³

2016

Af J Clin Exp Micro

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Noroviruses (NoV) are identified as the major cause of epidemic and sporadic acute gastroenteritis. Controlling the spread of the disease needs early recognition of NoV. This study investigated the contribution of norovirus to sporadic cases of pediatric gastroenteritis in Zagazig University Hospitals and studied the performance characteristics of enzyme linked immunosorbent assay(EIA) and immunochromatographic (ICT) assay for their ability to detect NoV. Two hundred stool specimens were collected from pediatric patients with acute gastroenteritis. Samples were tested for Norovirus presence by reverse transcription PCR (RT-PCR), ICT kit and EIA. 27% of the samples showed the 338-bp portion of the RNA-dependent RNA polymerase (RdRp) gene of both Norovirus genogroups I and II by RT-PCR. The ICT assay showed high specificity (97.94%) and high sensitivity (85.18%). The EIA showed high specificity (93.8%) but low sensitivity (64.8%). In conclusion, the high detection rate of NoV as the cause of diarrhea in children reported in this study supports their addition in screenings to identify sporadic cases of acute gastroenteritis. The ICT and RIA Norovirus kits may be useful for rapid screening of stool samples from patients with acute gastroenteritis. However, RT-PCR should be considered for negative samples to be confirmed. CHARACTERISTIQUES DE PERFORMANCE DU DOSAGE IMMUNO -ENZYMATIQUE ET TEST IMMUNOCHROMATOGRAPUIQUE RAPIDE POUR LE DEPISTAGE ROUTINE DU NOROVIRUS HUMAINE

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“…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].…”

Section: Introductionmentioning

confidence: 99%

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

Broglie¹,

Moore²,

Jaykus³

et al. 2014

J Anal Bioanal Tech

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This study designed and evaluated three versatile immuno-based assays for the rapid detection of GI.1 norovirus virus-like particles at low (0-3.0 μg/mL) levels: 1) enzymatic absorbance-based ELISAs, 2) a fluorescentbased immunoassay, and 3) a "signal-down" capture ELISA. Variables including controllable variations in assay format (indirect or sandwich), assay time, binding sequence, and reporter molecule (fluorophore or enzyme) were thoroughly investigated and optimized in all three assays. Selectivity tests in the three-hour absorbance-based ELISA using VLPs representing two GI and two GII strains indicated the assays were selective to GI strains over GII strains. The three-hour enzymatic absorbance-based assay turned out to be a robust and rapid method capable of detecting GI.I VLPs in the range of 0.037 to 0.555 μg/mL, and the three-hour fluorescent immunoassay was capable of detecting VLPs in a high concentration range of 0.5-2.0 μg/mL under optimized conditions. The "signal-down" capture ELISA was conceptually demonstrated for the detection of VLPs at concentrations in excess of 1.0 μg/ mL, but did not appear to be suitable for quantifying VLPs under its current conditions. The methods reported here provide proof-of-concept that various ELISA-type approaches could be further developed to provide robust norovirus detection assays having various detection ranges, limits, and linearity.

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Evaluation of a commercial ELISA for detecting Norwalk-like virus antigen in faeces

Cited by 96 publications

References 19 publications

Apparently non-specific results found using a norovirus antigen immunoassay for fecal specimens from neonates

Apparently non-specific results found using a norovirus antigen immunoassay for fecal specimens from neonates

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

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

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