Genus-specific kinetoplast-DNA PCR and parasite culture for the diagnosis of localised cutaneous leishmaniasis: applications for clinical trials under field conditions in Brazil

Abstract

The positivities of two methods for the diagnosis of localised cutaneous leishmaniasis (CL)

Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identificatio nof the parasite species

“…The capacity to detect Leishmania spp was compared with the PCR-mkDNA results. Although employing PCR-RFLP mkDNA for species identification appears to be inappropriate (see above), the high sensitivity of this method justifies its use for the molecular diagnosis of leishmaniasis (Pereira et al 2008, Ampuero et al 2009, Kobets et al 2010, Azmi et al 2011.…”

“…Whereas PCR assays targeted at amplification of internal transcribed spacer (ITS)1-rDNA are among the most commonly used methods for the diagnosis and identification of Leishmania species in the Old World (Schönian et al 2010a), PCR targeted at amplification of the conserved locus of kDNA minicircles (mkDNA) has been widely used in Brazil (Ampuero et al 2009). Digestion of the ITS1 amplicon using the restriction enzyme HaeIII has been demonstrated to be able to distinguish between nearly all Leishmania species, although distinction of some species, such as L. braziliensis and L. guyanensis, is only possible via sequence analysis of this region (Schönian et al 2003).…”

Three new sensitive and specific heat-shock protein 70 PCRs for global Leishmania species identification

“…Diagnosis of leishmaniasis currently focuses on parasite detection by microscopy, culture, and PCR. Molecular targets are either kinetoplast DNA (kDNA) [8] or genomic DNA (e.g. 18S rDNA [9]).…”

Specificity of the rapid rK39 antigen-based immunochromatographic test Kalazar Detect(r) in patients with cutaneous leishmaniasis in Brazil

“…The LCL diagnosis was confirmed by parasite culture or by kDNA detection with PCR using material that was obtained from ulcerated lesions, as previously reported (Ampuero et al 2009). The patients were from the south mesoregion of the state of Bahia, Brazil, an area that is endemic for L. brazi-liensis (Rosa et al 1988, Romero et al 2001).…”