Abstract:
BACKGROUND:Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol. METHODS/PRINCIPAL FINDINGS:Exploring the High Resolution Melting (HRM) dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR) targeting heat-shock protein 70 coding gene (hsp70) revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania) infantum chagasi, L. (L.) amazonensis, L. (L.) mexicana, L. (Viannia) lainsoni, L. (V.) braziliensis, L. (V.) guyanensis, L. (V.) naiffi and L. (V.) shawi, and three species found in Eurasia and Africa, including L. (L.) tropica, L. (L.) donovani and L. (L.) major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol. CONCLUSIONS/SIGNIFICANCE:HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA.
journal_name
PLoS Negl Trop Disjournal_title
PLoS neglected tropical diseasesauthors
Zampieri RA,Laranjeira-Silva MF,Muxel SM,Stocco de Lima AC,Shaw JJ,Floeter-Winter LMdoi
10.1371/journal.pntd.0004485subject
Has Abstractpub_date
2016-02-29 00:00:00pages
e0004485issue
2eissn
1935-2727issn
1935-2735pii
PNTD-D-15-01694journal_volume
10pub_type
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