BACKGROUND: Cryptosporidiosis is a significant cause of diarrheal illness in both immunocompetent and immunocompromised populations. Cryptosporidium species infect a wide range of hosts including humans. Different species are morphologically indistinguishable, and molecular techniques have become the key to detection and source tracking. The present study was designed to study the genetic diversity of human Cryptosporidium isolates in North India. METHODS: Cryptosporidium oocysts were detected in stool samples by special staining of fecal smears. DNA was extracted with a Qiagen kit and all samples were genotyped by small subunit ribosomal ribonucleic acid (SSU rRNA)-based nested PCR-restriction fragment length polymorphism (RFLP) tool using enzymes SspI and VspI. Cryptosporidium hominis and Cryptosporidium parvum isolates were subtyped by sequence analysis of the nested PCR amplified gp60 gene. RESULTS: Fifty-three fecal samples were found to be positive for Cryptosporidium oocysts. RFLP analysis revealed 39 isolates as C. hominis and 13 isolates of C. parvum; one sample failed amplification. gp60-based sequencing of C. hominis and C. parvum divided them into eight subgenotype families and 17 subtypes. gp60-based sequencing identified seven cases of mixed infection with C. hominis and C. parvum/Cryptosporidium meleagridis and showed the presence of C. meleagridis in six HIV-positive patients that were indistinguishable in RFLP. CONCLUSIONS: Cryptosporidium isolates obtained in the present study from patients in North India belonged to three species, eight subgenotype families, and 17 subtypes. The existence of many Cryptosporidium species, subgenotypes, and subtypes along with mixed infections reveals the complexity of Cryptosporidium transmission; this heterogeneity indicates stable cryptosporidiosis transmission in North India. The results may have further implications in understanding the epidemiology and control of this infection.
BACKGROUND: Cryptosporidiosis is a significant cause of diarrheal illness in both immunocompetent and immunocompromised populations. Cryptosporidium species infect a wide range of hosts including humans. Different species are morphologically indistinguishable, and molecular techniques have become the key to detection and source tracking. The present study was designed to study the genetic diversity of humanCryptosporidium isolates in North India. METHODS:Cryptosporidium oocysts were detected in stool samples by special staining of fecal smears. DNA was extracted with a Qiagen kit and all samples were genotyped by small subunit ribosomal ribonucleic acid (SSU rRNA)-based nested PCR-restriction fragment length polymorphism (RFLP) tool using enzymes SspI and VspI. Cryptosporidium hominis and Cryptosporidium parvum isolates were subtyped by sequence analysis of the nested PCR amplified gp60 gene. RESULTS: Fifty-three fecal samples were found to be positive for Cryptosporidium oocysts. RFLP analysis revealed 39 isolates as C. hominis and 13 isolates of C. parvum; one sample failed amplification. gp60-based sequencing of C. hominis and C. parvum divided them into eight subgenotype families and 17 subtypes. gp60-based sequencing identified seven cases of mixed infection with C. hominis and C. parvum/Cryptosporidium meleagridis and showed the presence of C. meleagridis in six HIV-positivepatients that were indistinguishable in RFLP. CONCLUSIONS:Cryptosporidium isolates obtained in the present study from patients in North India belonged to three species, eight subgenotype families, and 17 subtypes. The existence of many Cryptosporidium species, subgenotypes, and subtypes along with mixed infections reveals the complexity of Cryptosporidium transmission; this heterogeneity indicates stable cryptosporidiosis transmission in North India. The results may have further implications in understanding the epidemiology and control of this infection.
Authors: Jorge Néstor Velásquez; María Laura Pantano; Natalia Vittar; Mónica Gabriela Nigro; Olga Figueiras; Osvaldo Germán Astudillo; Javier Ricart; Daniela Della Paolera; Silvana Carnevale Journal: Parasitol Res Date: 2018-02-06 Impact factor: 2.289
Authors: Regina Helena Saramago Peralta; Jorge Néstor Velásquez; Flavia de Souza Cunha; María Laura Pantano; Fernando Campos Sodré; Sidnei da Silva; Osvaldo Germán Astudillo; José Mauro Peralta; Silvana Carnevale Journal: Mem Inst Oswaldo Cruz Date: 2016-01 Impact factor: 2.743