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

doi:10.1016/0022-1759(85)90065-1

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 journal › Article › peer-review

29 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 language	English (US)
Pages (from-to)	125-133
Number of pages	9
Journal	Journal of Immunological Methods
Volume	83
Issue number	1
DOIs	https://doi.org/10.1016/0022-1759(85)90065-1
State	Published - Oct 24 1985
Externally published	Yes

Keywords

amebiasis
axenic Entamoeba histolytica
dot ELISA

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1016/0022-1759(85)90065-1

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title = "A dot enzyme-linked immunosorbent assay for detection of antibodies against Entamoeba histolytica",

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.",

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AU - Kumar, Sanjai

AU - Band, A. H.

AU - Samantaray, J. C.

AU - Dang, N.

AU - Talwar, G. P.

PY - 1985/10/24

Y1 - 1985/10/24

N2 - 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.

AB - 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.

KW - amebiasis

KW - axenic Entamoeba histolytica

KW - dot ELISA

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

Abstract

Keywords

ASJC Scopus subject areas

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