A dot enzyme-linked immunosorbent assay for detection of antibodies against Entamoeba histolytica

Sanjai Kumar, A. H. Band, J. C. Samantaray, N. Dang, G. P. Talwar

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

A visual micro-dot enzyme-linked immunosorbent assay (ELISA) based on detection of antibodies against soluble antigens of axenically grown cultures of Entamoeba histolytica is described. The antigen was spotted on a nitrocellulose sheet, the unsaturated sites blocked with bovine serum albumin (BSA) and incubated with 3-fold dilutions of patient sera followed by incubation with protein A conjugated to peroxidase. Enzymic activity was evidenced using the substrate 4-chloro-1-naphthol. A positive reaction produced a blue spot. The sensitivity of the assay was better and comparable to the indirect haemag-glutination assay (IHA) and plate ELISA. The entire assay could be completed within 3 h. Antigen-loaded and pre-blocked nitrocellulose strips could be stored up to 3 months at room temperature and 37°C.

Original languageEnglish (US)
Pages (from-to)125-133
Number of pages9
JournalJournal of Immunological Methods
Volume83
Issue number1
DOIs
StatePublished - Oct 24 1985

Keywords

  • amebiasis
  • axenic Entamoeba histolytica
  • dot ELISA

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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