Data/hora: 11/06/2024 08:16:15

Provedor de dados: 	55	
País: 	Brazil	
Título: 	Design and evaluation of an AFLP molecular marker for the detection of Coccidioides spp. in biological samples	
Autores: 	Reyes-Montes,María del Rocío; Frías-De-León,María Guadalupe; Victoriano-Pastelín,Isai; Acosta-Altamirano,Gustavo; Duarte-Escalante,Esperanza.	
Data: 	2019-10-01	
Ano: 	2019	
Palavras-chave: 	Molecular markers; SCAR; Coccidioidomycosis; Coccidioides spp.	
Resumo: 	ABSTRACT At present, there is no standardized marker that is routinely used in clinical laboratories to diagnose coccidioidomycosis. Thus, the goals of this study were to obtain a sequence characterized amplified region (SCAR) marker for the identification of Coccidioides spp., evaluate its specificity and sensitivity in fungal DNA-spiked blood and sputum samples, and compare it with previously described molecular markers. Specific amplified fragment length polymorphism (AFLP) amplicons for Coccidioides immitis and Coccidioides posadasii were cloned into the vector pGEM® -T Easy vector and sequenced to develop a SCAR marker. Oligonucleotides were designed to identify Coccidioides spp. by polymerase chain reaction (PCR), and the specificity and sensitivity of these oligonucleotides were tested with the DNA from related pathogens. The specificity and sensitivity of the SCAR marker was evaluated with blood and sputum samples spiked with Coccidioides DNA and compared with other previously described markers (621, GAC2, and Ag2/PRA). In addition, the conditions for its use were established using biological samples. A specific marker named SCAR300 was obtained to identify Coccidioides spp. that exhibited good sensitivity and specificity. The results showed that all of the markers tested in this study can identify Coccidioides spp. However, the SCAR300 and 621 markers were the most sensitive, whereas the SCAR300 marker was the most specific. Thus, the SCAR300 marker is a useful tool to identify Coccidioides spp.	
Tipo: 	Info:eu-repo/semantics/article	
Idioma: 	Inglês	
Identificador: 	http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702019000500322	
Editor: 	Brazilian Society of Infectious Diseases	
Relação: 	10.1016/j.bjid.2019.08.002	
Formato: 	text/html	
Fonte: 	Brazilian Journal of Infectious Diseases  v.23 n.5 2019	
Fonte: 	info:eu-repo/semantics/openAccess