A real-time polymerase chain reaction assay for the identification and quantification of American Leishmania species on the basis of glucose-6-phosphate dehydrogenase


Autoria(s): CASTILHO, Tiago Moreno; CAMARGO, Luis Marcelo Aranha; MCMAHON-PRATT, Diane; SHAW, Jeffrey Jon; FLOETER-WINTER, Lucile Maria
Contribuinte(s)

UNIVERSIDADE DE SÃO PAULO

Data(s)

20/10/2012

20/10/2012

2008

Resumo

A real-time polymerase chain reaction (PCR) test was developed on the basis of the Leishmania glucose-6-phosphate dehydrogenase locus that enables identification and quantification of parasites. Using two independent pairs of primers in SYBR-Green assays, the test identified etiologic agents of cutaneous leishmaniasis belonging to both subgenera, Leishmania (Viannia) and Leishmania (Leishmania) in the Americas. Furthermore, use of TaqMan probes enables distinction between L. (V.) braziliensis or L. (V.) peruviania from the other L. (Viannia) species. All assays were negative with DNA of related trypanosomatids, humans, and mice. The parasite burden was estimated by normalizing the number of organisms per total amount of DNA in the sample or per host glyceraldehyde-3-phosphate dehydrogenase copies. The real-time PCR assay for L. (Leishmania) subgenus showed a good linear correlation with quantification on the basis of a limiting dilution assay in experimentally infected mice. The test successfully identifies and quantifies Leishmania in human biopsy specimens and represents a new tool to study leishmaniasis.

Identificador

AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, v.78, n.1, p.122-132, 2008

0002-9637

http://producao.usp.br/handle/BDPI/28572

http://apps.isiknowledge.com/InboundService.do?Func=Frame&product=WOS&action=retrieve&SrcApp=EndNote&UT=000252296700023&Init=Yes&SrcAuth=ResearchSoft&mode=FullRecord

Idioma(s)

eng

Publicador

AMER SOC TROP MED & HYGIENE

Relação

American Journal of Tropical Medicine and Hygiene

Direitos

restrictedAccess

Copyright AMER SOC TROP MED & HYGIENE

Palavras-Chave #CUTANEOUS LEISHMANIASIS #PCR ASSAY #KINETOPLAST DNA #QUANTIFYING LEISHMANIA #RAPID IDENTIFICATION #VIANNIA BRAZILIENSIS #ROBOTIC WORKSTATION #CLINICAL SPECIMENS #MOLECULAR BEACONS #INFECTED MICE #Public, Environmental & Occupational Health #Tropical Medicine
Tipo

article

original article

publishedVersion