Comparison of Polymerase Chain Reaction-restriction Fragment Length Polymorphism and Assemblage-specific Primers for Characterization of in Children
Overview
Affiliations
Background: is a diarrheagenic eukaryotic parasite that consists of at least eight morphologically identical but genetically distinct genotypes. Human giardiasis is caused mainly by A and B assemblages.
Aim And Objectives: The study aimed to compare the performance of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and assemblage-specific primers in genotyping of .
Materials And Methods: Stool samples of 315 children were microscopically screened for . Positive samples were genotyped using assemblage-specific primers and semi-nested PCR-RFLP techniques.
Results: The prevalence of was 18.1%. The detected genotypes using and approaches were assemblage A (15.8% vs. 12.7%) and assemblage B (36.8% vs. 74.5%) as single infections and mixed assemblages A and B (47.4% vs. 12.7%). The two approaches showed a moderate agreement (kappa index = 0.413, < 0.001). PCR-RFLP of gene revealed that sub-assemblages BIII and BIV were equally detected (30.9% each). The remaining samples were equally divided between sub-assemblage AII, mixed BIII and BIV, and mixed AII and BIII (12.7% each). A significant association was detected between the retrieved sub-assemblages and the presence of symptoms.
Conclusions: Although both approaches confirmed the predominance of assemblage B, the use of assemblage-specific primers is more effective in elucidating the true picture of mixed assemblage infection.
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