Analysis of nonspecific reactions in enzyme-linked immunosorbent assay testing for human rotavirus

R. H. Yolken, P. J. Stopa

Research output: Contribution to journalArticlepeer-review

Abstract

Solid-phase enzyme immunoassays can be utilized to detect antigens directly in clinical specimens. However, a small number of stools which we tested for human rotavirus by enzyme-linked immunosorbent assay (ELISA) were found to have nonspecific activity in the absence of rotaviral antigen. Similar nonspecific activity was found in eight of eight sera which contained rheumatoid factor. This nonspecific activity was markedly reduced by pretreatment of the specimens with reducing agents, normal goat serum, and anti-human immunoglobulin M (IgM). Thus, it is likely that these specimens contain an IgM antibody capable of reacting nonspecifically with the other components of the assay. Although pretreatment with the mild reducing agent N-acetylcysteine markedly reduced this nonspecific activity, such treatment did not reduce the specific ELISA activity due to rotarivus. Other treatments did produce a reduction in specific activity. Thus pretreatment with N-acetylcysteine offers a practical means to increase the specificity of ELISA systems without reducing their sensitivity.

Original languageEnglish (US)
Pages (from-to)703-707
Number of pages5
JournalJournal of clinical microbiology
Volume10
Issue number5
DOIs
StatePublished - 1979

ASJC Scopus subject areas

  • Microbiology (medical)

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