Genotypic identification of Cryptosporidium spp. isolated from HIV-infected patients and immunocompetent children of São Paulo, Brazil.
ABSTRACT Cryptosporidium isolates identified in fourteen stool samples, collected from five HIV-infected patients and nine immunocompetent children, living in the State of São Paulo, Brazil, were submitted to a molecular analysis using a nested PCR followed of restriction fragment length polymorphism (RFLP), for genetic characterization. The analysis was based on digestion with RsaI restriction enzyme of a DNA fragment amplified from the Cryptosporidium oocyst wall protein (COWP) gene. Based on this analysis, four samples were identified as Cryptosporidium parvum, eight as Cryptosporidium hominis and two presented a profile that corresponded to Cryptosporidium meleagridis when compared to the standards used in the analysis. The use of molecular methods can be helpful to identify source of infections and risk factors related to Cryptosporidium infection in our communities.
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ABSTRACT: Cryptosporidium spp. and Cystoisospora belli are monoxenic protozoa that have been recognized as the causative agents of chronic diarrhea in immunocompromised individuals, especially HIV-infected subjects. The objective of this study was to evaluate the frequency of these intestinal protozoa in HIV-positive patients in the Triângulo Mineiro region of Brazil and to correlate the presence of these infections with clinical, epidemiological and laboratory data of the patients. Oocysts were detected in stool samples of 10 (16.9%) of the 59 patients studied, while Cryptosporidium spp. were present in 10.1% (6/59) and C. belli in 6.7% (4/59). The frequency of these parasites was higher among patients with diarrheic syndrome and CD4+ T lymphocyte counts < 200 cells/mm 3 , demonstrating the opportunistic characteristic of these infections. A significant association was observed between the lack of adherence to antiretroviral therapy and the presence of Cryptosporidium spp. and/or C. belli. Parasitism with Cryptosporidium spp. was more frequent in February and April, the months following the period of high rainfall. The same was not observed for C. belli. Genetic characterization of two isolates led to the identification of Cryptosporidium parvum, one of the main species associated with the zoonotic transmission of cryptosporidiosis.Revista do Instituto de Medicina Tropical de São Paulo 05/2013; 55(3). DOI:10.1590/S0036-46652013000300002 · 0.91 Impact Factor
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ABSTRACT: Objetivo: Caracterizar genéticamente Cryptosporidium spp para determinar la diversidad de especies en seres humanos. Pacientes y Métodos: estudio transversal realizado en Valparaíso, Chile, Un total de 458 pacientes participaron del estudio; 259 inmunodeficientes (pacientes con infección por VIH, oncológicos, con trasplante renal, síndrome de hiper IgM y una mujer embarazada sin infección por VIH) y 178 inmunocompetentes proporcionaron muestras fecales y 21 muestras de bilis. Resultados: Se obtuvieron 29 (6,3%) muestras positivas; 25 (9,7%) de inmunodeficientes: 18 (7,3%) de pacientes con infección por VIH y 7 con otras inmunodeficiencias; los restantes 4 (2,2%) fueron de personas inmunocompetentes. La genotipificación de Cryptosporidium se efectuó mediante reacción de polimerasa en cadena (RPC) anidada y el polimorfismo de la longitud de los fragmentos de restricción y/o RPC - secuenciación de la SSU ARNr, a partir de ooquistes en la muestra fecal. Se identificaron 4 especies: C. parvum, C. hominis, C. muris y C. meleagridis. En pacientes inmunodeficientes, se caracterizaron 16 C. parvum, 8 C. hominis y un C. muris; en inmunocompetentes: 3 C. hominis y un C. meleagridis. Conclusión: Los resultados indican que se produce transmisión zoonótica y antroponótica y que C. parvum es la especie predominante en este estudio. Las especies de Cryptosporidium de transmisión zoonótica representan el 62% en los seres humanos participantes de este estudio.Revista chilena de infectologia: organo oficial de la Sociedad Chilena de Infectologia 02/2012; 29(1):63-71. DOI:10.4067/S0716-10182012000100011 · 0.50 Impact Factor
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ABSTRACT: Objectives. The prevalence of Cryptosporidium and the risk factors of zoonotic transmission in Najafabad, Isfahan, Iran dairy farms were examined. Methods. One fecal sample was collected from all calves less than 6 months old in eight dairy farms around Najafabad (Isfahan province, Central Iran) as well as individuals working in these farms and their household members. A two-step nested PCR protocol was used to amplify the 18S rRNA gene (830 bp). Results. Cryptosporidium was identified in the stool of 36 (prevalence 8.5%) of 96 farm workers and 326 household members. Furthermore, 31 (14.2%) of 218 calf samples were positive. Cryptosporidium parvum was identified in 15 (72%) of the positive farm workers and 10 (65%) of the positive household members. Of the positive calves, 20 (64.5%) were infected with C. parvum, indicating possible zoonotic transmission in these farms. Contact with calves (P < 0.0001) was the most significant risk factor of C. parvum infection. A considerable negative association was observed between C. parvum infection and cleaning of shoes/boots after daily work (P = 0.004), hand washing (P = 0.013), and use of piped water (P < 0.006). In the multivariate analysis with logistic regression, only contact with calves was significant. Conclusion. Zoonotic transmission of C. parvum due to contact with calves is predominant among farm workers and their household members of this region and appropriate health measures must be applied to control the infection and decrease of zoonotic transmission of this parasite.Journal of Tropical Medicine 04/2014; 2014:405875. DOI:10.1155/2014/405875