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

• Published in: Journal of clinical virology 2003, Vol.26 (1), p.109-115
• Main Authors: Richards, A.F, Lopman, B, Gunn, A, Curry, A, Ellis, D, Cotterill, H, Ratcliffe, S, Jenkins, M, Appleton, H, Gallimore, C.I, Gray, J.J, Brown, D.W.G
• Format: Article
• Language: English
• Subjects: Antigen detection; Antigens, Viral - analysis; Biological and medical sciences; Caliciviridae Infections - epidemiology; Caliciviridae Infections - virology; Capsid Proteins - analysis; Capsid Proteins - genetics; Capsid Proteins - immunology; Disease Outbreaks; EIA; Enzyme-Linked Immunosorbent Assay; Feces - virology; Fundamental and applied biological sciences. Psychology; Gastroenteritis - epidemiology; Gastroenteritis - virology; Genotype; Human viral diseases; Humans; Infectious diseases; Medical sciences; Microbiology; Microscopy, Electron; NLV; Norovirus; Norovirus - classification; Norovirus - genetics; Norovirus - immunology; Norovirus - isolation & purification; Reagent Kits, Diagnostic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral - genetics; Sensitivity and Specificity; SRSV; Stools; Techniques used in virology; Viral diseases; Viral diseases of the digestive system; Virology
• ISSN: 1386-6532; 1873-5967
• DOI: 10.1016/S1386-6532(02)00267-6

A commercially available enzyme immunoassay, the IDEIA™ Norwalk-like virus (NLV) enzyme linked immunosorbent assay (ELISA; Dako Cytomation, Ely, UK) for detecting NLV antigen in faecal samples and determining the NLV genogroup was evaluated. The performance of the ELISA was compared with that of electron microscopy and the reverse transcription polymerase chain reaction by testing a panel of faecal samples collected from patients involved in outbreaks of gastroenteritis. When compared with reverse transcription-polymerase chain reaction (RT-PCR), the ELISA had a sensitivity and specificity of 55.5 and 98.3%, respectively. This compares with a sensitivity and specificity for EM of 23.9 and 99.2%, respectively. The sensitivity and specificity of the ELISA for determining the aetiology of a Norwalk virus-like outbreak, based on two or more positive samples within an outbreak, were 52.2 and 100% when two samples were collected from an outbreak and 71.4 and 100% when six or more samples were collected. The ELISA correctly identified the NLV genogroups of viruses previously characterised by partial DNA sequencing. The ELISA is a suitable alternative to the preliminary screening by EM for investigating outbreaks of gastroenteritis. Outbreaks, negative by ELISA should be examined by RT-PCR in order to detect strains non-reactive in the assay and virus strains from representative ELISA positive outbreaks should be characterised fully to allow the genetic diversity of NLVs co-circulating in the population to be described.