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Human cysticercosis and taeniasis: molecular approaches for specific diagnosis and parasite identification
D P McManus1, E Garcia-Zepeda, A Reid
1Department of Pure and Applied Biology, Imperial College of Science and Technology, London, U.K.
Abstract:
The construction and antibody screening of Taenia solium cDNA libraries, generated in the Escherichia coli bacteriophage lambda gt11, with the identification of clones putatively expressing antigen B, T. solium-specific and other antigens is described. Lysogens were produced from a number of selected clones and beta-galactosidase fusion peptides ranging in Mr of approximately 135,000-150,000 were demonstrated. These proteins were shown by immunoblotting to be reactive with a pool of sera from cysticercotic patients originally used in the cDNA library screening. We report a method whereby Taenia (T. saginata and T. pisiformis) eggs can be detected with high sensitivity in a specific DNA dot-blot hybridisation assay using total parasite DNA as probe. We show also that intra-specific DNA variability occurs in T. solium isolates obtained from different geographical areas and discuss the potential significance of this heterogeneity.
Insights
Researchers screened Taenia solium cDNA libraries to identify antigens, aiding in cysticercosis diagnosis. A sensitive DNA dot-blot assay was developed for detecting Taenia eggs, revealing genetic diversity in Taenia solium populations.
Area of Science:
- Molecular Biology
- Parasitology
- Immunology
Background:
- Taenia solium causes cysticercosis, a significant human health concern.
- Accurate detection of Taenia species and their antigens is crucial for diagnosis and control.
Purpose of the Study:
- To construct and screen Taenia solium cDNA libraries for antigen identification.
- To develop a sensitive DNA-based method for detecting Taenia eggs.
- To investigate genetic variability within Taenia solium isolates.
Main Methods:
- Construction and antibody screening of Escherichia coli bacteriophage lambda gt11 cDNA libraries.
- Production of lysogens and demonstration of beta-galactosidase fusion peptides.
- Immunoblotting to confirm reactivity of fusion proteins with patient sera.
- Development of a DNA dot-blot hybridization assay using total parasite DNA as a probe.
Main Results:
- Identification of Taenia solium clones expressing specific antigens, including antigen B.
- Demonstration of reactive beta-galactosidase fusion peptides in selected clones.
- High sensitivity achieved in detecting Taenia eggs using the DNA dot-blot assay.
- Evidence of intra-specific DNA variability in Taenia solium from different geographical regions.
Conclusions:
- cDNA library screening is effective for identifying Taenia solium antigens.
- The developed DNA dot-blot assay offers a sensitive method for Taenia egg detection.
- Genetic heterogeneity in Taenia solium populations may have epidemiological significance.