Data/hora: 10/06/2024 18:41:26

Provedor de dados: | 58
País: | Brazil
Título: | Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples
Autores: | Miotto,Bruno Alonso; Hora,Aline Santana da; Taniwaki,Sueli Akemi; Brandão,Paulo Eduardo; Heinemann,Marcos Bryan; Hagiwara,Mitika Kuribayashi.
Data: | 2018-09-01
Ano: | 2018
Palavras-chave: | Leptospirosis; Dog; Urine; QPCR; Lipl32.
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.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000300584
Editor: | Sociedade Brasileira de Microbiologia
Relação: | 10.1016/j.bjm.2017.09.004
Formato: | text/html
Fonte: | Brazilian Journal of Microbiology  v.49 n.3 2018
Fonte: | info:eu-repo/semantics/openAccess