First Molecular Characterization of Cryptosporidium spp. in Patients Living with HIV in Honduras

Abstract

Cryptosporidiosis is one of the most important causes of gastroenteritis in the world, especially in low- and middle-income countries. It is caused by the Apicomplexan parasite Cryptosporidium spp., and mainly affects children and immunocompromised people, in whom it can pose a serious threat to their health, or even be life threatening. In Honduras, there are no data on parasite species or on molecular diversity or Cryptosporidium subtypes. Therefore, a cross-sectional study was conducted between September 2019 and March 2020 for the molecular identification of Cryptosporidium spp. in 102 patients living with HIV who attended a national hospital in Tegucigalpa. Stool samples were analyzed by direct microscopy, acid-fast stained smears, and a rapid lateral flow immunochromatographic test. All samples that tested positive were molecularly analyzed to identify the species and subtype of the parasite using three different markers: gp60, cowp, and 18Sr. PCR products were also sequenced. Four out of 102 samples (3.92%) were positive for Cryptosporidiumparvum, and all were assigned to subtype IIa. These findings suggest a possible zoonotic transmission in this population.

Keywords: 18S ribosomal; C. parvum; Cryptosporidium spp; HIV/AIDS; Honduras; cowp; gp60.

Figure 1

Geographical origin of the study participants.

Figure 2

Source of drinking water of the study participants.

Figure 3

Number and species of intestinal parasites found in the participating individuals.

Figure 4

Agarose electrophoresis showing: (a) PCR product of the gp60 gene; (b) PCR amplification of the cowp gene; (c) Digestion of the PCR product of the cowp gene using RsaI; (d) PCR amplification of the 18S ribosomal fragment.

Figure 5

Working algorithm of three molecular markers for the identification of Cryptosporidium spp. (a) Nested PCR of the gp60 gene [19]; (b) Nested PCR of the cowp gene, and PCR-RFLP for species identification [24]; (c) Nested PCR of the 18S ribosomal gene [10].