A simple method for evaluating Cryptosporidium‐specific antibodies used in monitoring environmental water samples

A simple method for evaluating Cryptosporidium‐specific antibodies used in monitoring environmental water samples

A simple method is described for the evaluation and quality control of Cryptosporidium‐specific antibodies used in monitoring environmental water samples. Purified oocysts were fluorescently labelled with a test antibody at the appropriate concentration. Labelled oocysts were analysed using flow cyt...

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Bibliographic Details
Published in:Letters in applied microbiology Vol. 25; no. 5; pp. 316 - 320
Main Authors: Vesey, G., Deere, D., Weir, C.J., Ashbolt1, N., Williams, K.L., Veal, D.A.
Format: Journal Article
Language:English
Published: Oxford, UK Blackwell Science Ltd 01-11-1997
Blackwell Science
Subjects:
Animals
Antibodies, Protozoan - analysis
Biological and medical sciences
Biotechnology
Cryptosporidium - immunology
Flow Cytometry
Fluorescent Antibody Technique
Fundamental and applied biological sciences. Psychology
General aspects and techniques
Protozoa
Water - parasitology
Performance evaluation
Protozoa
Cryptosporidium
Flow cytometry
Apicomplexa
Parasite
Monoclonal antibody
Method
Optimization
Specificity
Quality control
Water quality
Detection
Immunofluorescence
Monitoring
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Summary:A simple method is described for the evaluation and quality control of Cryptosporidium‐specific antibodies used in monitoring environmental water samples. Purified oocysts were fluorescently labelled with a test antibody at the appropriate concentration. Labelled oocysts were analysed using flow cytometry and a region was defined on a bivariate dotplot of fluorescence versus light scatter that enclosed all oocysts. Concentrates of environmental water samples that did not contain oocysts were then incubated with the test antibody and analysed using flow cytometry. The number of particles that appeared in the region defined for oocysts was recorded and was a measure of non‐specific binding. The technique provides a simple, rapid and quantitative tool for both evaluating the binding specificity of test antibodies and optimizing sample staining conditions.
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ISSN:0266-8254
1472-765X
DOI:10.1046/j.1472-765X.1997.00237.x