Data/hora: 01/06/2024 05:42:08

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