title: Evaluation of end-point titration, single dilution and capture enzyme immunoassays for measurement of antirotaviral IgA and IgM in infantile secretions and serum.
abstract: In order to facilitate measurement of antirotaviral IgA in large collections of faeces and secretions, adaptations of enzyme immunoassay methods for estimating antirotaviral IgA and IgM in duodenal fluid, saliva, faeces and serum were studied. To quantitate specific IgA, a single dilution of each sample was assayed. Results were expressed as antirotaviral IgA units derived from a standard curve. Units were calculated by log-logit analysis on computer. There was strong correlation between antirotaviral IgA units and end-point titres in 257 faecal samples (correlation coefficient r = 0.92) and in 182 duodenal fluids and salivary samples (correlation coefficient r = 0.74). The assay was validated using acute and convalescent faeces from children with or without rotavirus infection. Immune conversions in IgA were detected in 33 (75%) of the children by units and 34 (77%) by titres. None of nine children with gastroenteritis due to other infectious agents showed immune conversions to rotavirus. A monoclonal capture IgM assay showed similar end-point titres and numbers of immune conversions when compared with a direct assay for antirotaviral IgM in serum and secretions. Use of the capture method eliminated false-positive reactions with the cell control. The assay for antirotaviral IgA units in secretions is simple, rapid, reproducible and reliable, and has proven of value in longitudinal epidemiological studies of rotavirus coproIgA profiles. Both the capture IgM technique and the single dilution IgA method permit analysis of large numbers of specimens and are appropriate for examination of immune responses to natural rotavirus infection or during vaccine trials.
authors.affiliation: Department of Gastroenterology, Royal Children's Hospital, Parkville, Victoria, Australia.
authors.forename: B S
authors.initials: BS
authors.lastname: Coulson
authors.forename: K
authors.initials: K
authors.lastname: Grimwood
authors.forename: R F
authors.initials: RF
authors.lastname: Bishop
authors.forename: G L
authors.initials: GL
authors.lastname: Barnes
mesh_terms: D000914:Antibodies, Viral / Q000032:analysis*; D002675:Child, Preschool; D005243:Feces / Q000032:analysis; D005759:Gastroenteritis / Q000097:blood / Q000276:immunology* / Q000382:microbiology; D005766:Gastrointestinal Contents / Q000032:analysis; D006801:Humans; D007124:Immunoenzyme Techniques; D007070:Immunoglobulin A / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis*; D007223:Infant; D007231:Infant, Newborn; D012401:Rotavirus / Q000276:immunology* / Q000302:isolation & purification; D012463:Saliva / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D007070:Immunoglobulin A; D007075:Immunoglobulin M