Evaluación de métodos moleculares y microscópicos para la detección de Cryptosporidium spp. (apicomplexa – cryptosporidiidae).

Abstract

Introduction. Cryptosporidiosis is an emerging disease caused by protozoa of  the  Cryptosporidium genus, which affects a wide range of vertebrates, including humansIts prevalence ranges from 4%-6% in Central and South America and can even cause death in immunosuppressed patients reason why it is considered a public health problem worldwide. It is necessary to implement and evaluate strategies for detection and typification of different Cryptosporidium species,to adopt control measures and monitoring. Objective. To perform a comparison between microscopic and molecular methods for the detection and typification of Cryptosporidium species with the purpose of using the most sensitive method in the detection of Cryptosporidium oocysts in water samples. Materials and methods. Detection and typification of Cryptosporidium spp, in feces and water samples were done initially using a concentration method for both feces and water (Formol-ether and inorganic flocculation method with Calcium carbonate); the parasite identification was carried out using the Ziehl-Nelseen staining and the Polymerase Chain Reaction PCR amplification of ribosomal ADN regions, of gens codified for Hsp70 protein and the gen that codifies for the Cryptosporidium oocyst wall protein (COWP). The typification was carried out by digestion with restriction enzymes SspI, RsaI and VspI. Results. The Ziehl-Neelsen staining, confirmed the presence of Cryptosporidium  spp., in 10 of the 168 samples tested (humans, calves, dogs and rabbits), the PCR typification, confirmed 15 positive samples for C. parvum and one for C. hominis. Conclusion. The sensitivity of detection of  Cryptosporidium by PCR and its utility in the diagnosis is shown, registering the presence of two species of the parasite circulating in samples taken in the municipality of Manizales.

Keywords:

Cryptosporidium spp.

rADN

Hsp70

COWP

Ziehl-Neelsen

Cryptosporidium spp.

rDNA

Hsp70

COWP

Ziehl-Neelsen

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