Ocorrência de papilomavírus humano na cérvice uterina de mulheres da região ocidental da Amazônia Brasileira / Occurrence of human papillomavirus in uterine cervix of women in the western Brazilian Amazon

The occurrence of human papillomavirus (HPV) is a public health problem, because it has been linked to cancer. The aim of this research was to identify the occurrence of human papillomavirus in uterine cervix of women in the western Brazilian Amazon. The study was conducted in the capital of Rondonia, Porto Velho. We identified the types of HPV, also we correlated molecular results with those of colpocytologic tests coming from 334 women who underwent preventive examinations at the Brazilian Unified Health System. From samples, we obtained genetic material of the human papillomavirus (HPV ADN) and amplified the 450-bp fragment from the conserved region of the L1 gene, which was submitted to restriction fragment length polymorphism (RFLP) analysis. Of the 334 samples analyzed, 31% were confirmed to have the presence of viral material (HPV ADN). We detected the existence of the following HPV types: 16, 18, 33, 53 and 58, which identifies the group of high oncogenic risk, with 72% (74/103) of occurrence. Also we found HPV types 11, 42 and 44, which belong to the group of low oncogenic risk, with 28% of occurrence. The recurrent profiles in the development of the analysis were HPV-16 and -18, with 17% and 16% respectively. We found that more than 80% of the samples that contained viral material did not show any abnormal cell in the cytology test. This finding reinforces the need to disseminate the use of molecular techniques in conventional diagnostics.

A ocorrência do papiloma vírus humano (HPV) é um problema de saúde pública, pois tem sido associado ao câncer. O objetivo da pesquisa foi identificar a ocorrência de papilomavírus humano na cérvice uterina de mulheres da região ocidental da Amazônia Brasileira. O estudo foi realizado na capital de Rondônia, Porto Velho. Foram identificados os tipos de HPV e resultados moleculares foram correlacionados com aqueles os testes colpocitológicos de amostras provenientes de 334 mulheres que realizaram exames preventivos no Sistema Único de Saúde. Obteve-se o material genético viral do papilomavírus humano (DNA-HPV) e o fragmento de 450 pb da região conservada do gene L1 amplificado e submetido à análise do polimorfismo dos fragmentos de restrição (RFLP). Das 334 amostras analisadas, 31% foram confirmados com a presença de material viral (DNA-HPV). Confirmou-se a existência dos tipos: HPVS-16, 18, 33, 53 e 58, que identificam o grupo de alto risco oncogênico com 72% (74/103) de ocorrência, bem como os HPVS-11, 42 e 44 pertencentes ao grupo de baixo risco oncogênico com 28% de ocorrência. Os perfis recorrentes durante o desenvolvimento da análise foram do HPV-16 e -18 com 17% e 16%, respectivamente. Os resultados da pesquisa indicam que mais de 80% das amostras analisadas e que continham material viral não apresentavam nenhuma alteração celular no teste citológico, o que reforça a necessidade de se difundir o uso das técnicas moleculares em diagnósticos convencionais.

Nontuberculous mycobacteria in respiratory samples from patients with pulmonary tuberculosis in the state of Rondonia, Brazil

The main cause of pulmonary tuberculosis (TB) is infection with Mycobacterium tuberculosis (MTB). We aimed to evaluate the contribution of nontuberculous mycobacteria (NTM) to pulmonary disease in patients from the state of Rondônia using respiratory samples and epidemiological data from TB cases. Mycobacterium isolates were identified using a combination of conventional tests, polymerase chain reaction-based restriction enzyme analysis of hsp65 gene and hsp65 gene sequencing. Among the 1,812 cases suspected of having pulmonary TB, 444 yielded bacterial cultures, including 369 cases positive for MTB and 75 cases positive for NTM. Within the latter group, 14 species were identified as Mycobacterium abscessus, Mycobacterium avium, Mycobacterium fortuitum, Mycobacterium intracellulare, Mycobacterium gilvum, Mycobacterium gordonae, Mycobacterium asiaticum, Mycobacterium tusciae, Mycobacterium porcinum, Mycobacterium novocastrense, Mycobacterium simiae, Mycobacterium szulgai, Mycobacterium phlei and Mycobacterium holsaticum and 13 isolates could not be identified at the species level. The majority of NTM cases were observed in Porto Velho and the relative frequency of NTM compared with MTB was highest in Ji-Paraná. In approximately half of the TB subjects with NTM, a second sample containing NTM was obtained, confirming this as the disease-causing agent. The most frequently observed NTM species were M. abscessus and M. avium and because the former species is resistant to many antibiotics and displays unsatisfactory cure rates, the implementation of rapid identification of mycobacterium species is of considerable importance.

Genotyping of Mycobacterium leprae present on Ziehl-Neelsen-stained microscopic slides and in skin biopsy samples from leprosy patients in different geographic regions of Brazil

We analysed 16 variable number tandem repeats (VNTR) and three single-nucleotide polymorphisms (SNP) in Mycobacterium leprae present on 115 Ziehl-Neelsen (Z-N)-stained slides and in 51 skin biopsy samples derived from leprosy patients from Ceará (n = 23), Pernambuco (n = 41), Rio de Janeiro (n = 22) and Rondônia (RO) (n = 78). All skin biopsies yielded SNP-based genotypes, while 48 of the samples (94.1%) yielded complete VNTR genotypes. We evaluated two procedures for extracting M. leprae DNA from Z-N-stained slides: the first including Chelex and the other combining proteinase and sodium dodecyl sulfate. Of the 76 samples processed using the first procedure, 30.2% were positive for 16 or 15 VNTRs, whereas of the 39 samples processed using the second procedure, 28.2% yielded genotypes defined by at least 10 VNTRs. Combined VNTR and SNP analysis revealed large variability in genotypes, but a high prevalence of SNP genotype 4 in the Northeast Region of Brazil. Our observation of two samples from RO with an identical genotype and seven groups with similar genotypes, including four derived from residents of the same state or region, suggest a tendency to form groups according to the origin of the isolates. This study demonstrates the existence of geographically related M. leprae genotypes and that Z-N-stained slides are an alternative source for M. leprae genotyping.