Multilocus Genotyping of Cryptosporidium Hominis Associated With Diarrhea Outbreak in a Day Care Unit in São Paulo

Abstract

Gonçalves EM do N, da Silva AJ, Eduardo MB de P, Uemura IH, Moura INS, Castilho VLP et al. Multilocus genotyping of Cryptosporidium hominis associated with diarrhea outbreak in a day care unit in São Paulo. Clinics. 2006;61(2):119-26.A number of species of Cryptosporidium are associated with diarrhea worldwide. Little data exists regarding the genotypes and species of Cryptosporidium associated with cases of infections in Brazil. PURPOSE: In the present study, we ascertained by molecular methods the species and the genotype of Cryptosporidium sp from a diarrhea outbreak diagnosed in a day care at the Hospital Clínicas, São Paulo University Medical School. MATERIALS AND METHODS: Specific identification and typing of the isolates associated with the outbreak was done by DNA sequencing analysis of fragments amplified by polymerase chain reaction (PCR) from 3 different Cryptosporidium loci: the SSUrRNA coding region, the Cryptosporidium oocyst wall protein (COWP) gene, and the microsatellite locus 1 (ML1), a tandem GAG-trinucleotide repeat containing substitutions that differentiate the genotypes of Cryptosporidium parvum and Cryptosporidium hominis. RESULTS: A total of 29 positive samples from the outbreak were studied by the molecular methods described. Our study revealed the presence of a single genotype of Cryptosporidium hominis in all samples. CONCLUSION: The molecular analysis reinforced the hypothesis that the transmission of Cryptosporidium hominis during the period the samples were collected occurred in an outbreak pattern, possibly by person-to-person contact through the fecal-oral route. As far as we know, this is the first time that molecular tools have been used to identify the species and the genotype of isolates showing the presence of the ML1 genotype in samples from Brazilian patients.

Prevalence and genetic characterization of Cryptosporidium spp. and Cystoisospora belli in HIV-infected patients

“…In Brazil, the prevalence of cryptosporidiosis and cystoisosporosis among HIV-infected patients ranges from 6.4% to 9.1% and from 4.4% to 18%, respectively 1,2,6,7,11,16,18 . OLIVEIRA-SILVA et al 19 investigated the frequency of these parasites in HIV-infected patients living in the Triângulo Mineiro region and found a prevalence of 8.6% for Cryptosporidium sp.…”

A multiattribute utility evaluation for the detection of Cryptosporidium species in stool

“…Cryptosporidium parvum (C. parvum) is associated with massive diarrhea outbreaks worldwide, generally caused by exposure to drinking or recreational water or direct contact with infected persons through the oral-fecal route [2,3]. Diarrhea is exceedingly common and exerts an enormous toll in terms of mortality, morbidity, loss of work productivity, consumption of medical resources and social inconvenience.…”

Genotypic identification of Cryptosporidium spp. isolated from hiv-infected patients and immunocompetent children of São Paulo, Brazil

“…was first registered in 1976, and Cryptosporidium parvum (previously known as cattle genotype or genotype 2) and Cryptosporidium hominis (previously known as C. parvum -human genotype or genotype 1) have been recorded as the two species most frequent cause of human cryptosporidiosis, either associated with sporadic cases of infections or outbreaks, some of them being very large as the one that occurred in Milwaukee WI, USA, which affected approximately 400,000 persons 12,17 . To date, all the cryptosporidiosis outbreaks occurring worldwide have been caused by C. hominis and C. parvum, with several being associated with consumption of drinking water or exposure to recreational water contaminated with Cryptosporidium oocysts of zoonotic and anthroponotic origins 15,27,30 . Due to the size and frequency of these outbreaks, cryptosporidiosis became a serious public health issue worldwide and prompted reevaluation of the microbiological standards for drinking water by health authorities in developed and developing countries.…”

“…in their stools 5,7,8,15,19,25 . Serological surveys conducted in Brazil showed the presence of antiCryptosporidium antibodies in a large number of children living in slums of the northeastern region of Fortaleza, state of Ceara 1 , and also in individuals with no intestinal symptoms living in the state of São Paulo.…”

“…In one of the studies carried out in the northeast region, two isolates were identified as C. parvum, one as C. hominis, but the authors were not able to genetically characterized one of the isolates 2 . Other study identified seven Brazilian children infected with C. hominis 14 and another revealed the presence of a single genotype of C. hominis based on analysis of three genes in 29 samples from a diarrhea outbreak investigated in a day care in São Paulo, SP 15 .…”