Detecção e quantificação de mycobacterium leprae pela técnica de pcr em tempo real em amostras ambientais de municípios do Ceará

• Autor(a) principal: Holanda, Maisa Viana de
• Data de Publicação: 2015
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
• Texto Completo: http://www.repositorio.ufc.br/handle/riufc/13593
• topic: Mycobacterium leprae; DNA; Ingestão de Líquidos

ABSTRACT Leprosy is a chronic granulomatous infectious disease occasioning damage in skin and peripheral nerves, caused by Mycobacterium leprae , an intr a cellular agent unable to grown in axenic culture media. In 2012 , 232.857 new cases were detected, and out of these 33.303 (14, 30%) were detected only in Brazil. Ceará diagnosed 2,066 new cases only in 2012, with an overall detection ratio of 24.0 / 100,000 i nhabitants . Transmission is related to the bacilli eliminate from MB patients by the upper respira tory route and consequent contamination the air . The existence of clinical cases in which patients had no previous contact with leprosy patients and also the finding of cases near water source s suggests infection of humans by environmental sources . C urrently, it is suggested the possible influence of environmental factors and wild animals in the transmission of leprosy as soil, water, vegetation and armadillos. The environment behave as a kind of reservoir, allowing infectious bacilli remain viable af ter long perio ds outside the body. This study aims to detect and quantify M. leprae bacillus by qPCR environmental water samples from Ceará municipalities . Five samples from each one of the 30 water reservoir were collected, being a total of 149 samples. Total DNA was extracted by specific environmental samples kit according to the manufacturer's recommendations. Subsequently, was performed the amplification of M. leprae 16S rRNA gene by qPCR using the SYBR Green PCR Master Mix kit. We used a standard curv e with known concentrations of pIDTBlue 16SrRNAMlep plasmid to quantify the DNA present in environmental samples. Environmental samples of Juazeiro do Norte municipality and clinical samples from patients treated at CDREM were genotyped and subtyped by PCR - RFLP. M. leprae ’s DNA was detected in all the districts studied. Out of the total of 149 w ater samples analyzed, 81 (54.4 %) were positive for the detection of DNA. The number of copies of M. leprae was in the range of 1,42 x 10 - 1 to 1,44 x 10 + 2 . Most clinical samples presented genotype 4 (64%) whereas 100% of Juazeiro samples were SNP 4. T he SNP 4 - N was the most present both among the clinical l and environmental samples. This study indicates the presence of M. leprae DNA in samples of en vironmental wate r, essentially demonstrates the presence of bacilli in the water analyzed . Thus, further studies are needed in order to enhance knowledge of the influence of the water in the transmission of leprosy.