| Texto completo | |
| Autor(es): |
Miotto, Bruno Alonso
[1]
;
da Hora, Aline Santana
[2]
;
Taniwaki, Sueli Akemi
[2]
;
Brandao, Paulo Eduardo
[2]
;
Heinemann, Marcos Bryan
[2]
;
Hagiwara, Mitika Kuribayashi
[1]
Número total de Autores: 6
|
| Afiliação do(s) autor(es): | [1] Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Clin Med, Sao Paulo, SP - Brazil
[2] Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Med Vet Prevent & Saude Anim, Sao Paulo, SP - Brazil
Número total de Afiliações: 2
|
| Tipo de documento: | Artigo Científico |
| Fonte: | Brazilian Journal of Microbiology; v. 49, n. 3, p. 584-590, JUL-SEP 2018. |
| Citações Web of Science: | 3 |
| Resumo | |
Abstract A modified TaqMan real-time polymerase chain reaction targeting a 138 bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleotide sequencing analysis. The overall agreement between the assays was 94.8% (confidence interval [CI] 95% 88-100%). The newly developed assay presented 91.6% (CI 95% 71.5-98.5%) relative sensitivity (22[+] lipl32 PCR/24[+] 16S RNA and sequencing), 100% (CI 95% 96.3-100%) relative specificity and 98.7% accuracy (CI 95% 94.8-100%). The lipl32 assay was able to detect and quantify at least 10 genome equivalents/reaction. DNA extracted from 17 pathogenic Leptospira spp., 8 intermediate/saprophytic strains and 21 different pathogenic microorganisms were also tested using the lipl32 assay, resulting in amplification exclusively for pathogenic leptospiral strains. The results also demonstrated high intra and inter-assay reproducibility (coefficient of variation 1.50 and 1.12, respectively), thereby qualifying the newly developed assay as a highly sensitive, specific and reliable diagnostic tool for leptospiral infection in dogs using urine specimens. (AU) | |
| Processo FAPESP: | 12/14681-7 - Avaliação da leptospirúria e identificação de portadores de leptospiras patogênicas em cães mantidos em abrigos públicos ou particulares da região metropolitana de São Paulo |
| Beneficiário: | Mitika Kuribayashi Hagiwara |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 12/13022-0 - Avaliação da leptospirúria e identificação de portadores de leptospiras patogênicas em cães mantidos em abrigos públicos ou particulares da Região Metropolitana de São Paulo |
| Beneficiário: | Bruno Alonso Miotto |
| Modalidade de apoio: | Bolsas no Brasil - Mestrado |