New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction

通过聚合酶链式反应检测和区分利什曼原虫和利什曼原虫亚属的新引物

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作者:Manuel Calvopiña, David Fonseca-Carrera, Irina Villacrés-Granda, Alberto Toapanta, Carlos Chiluisa-Guacho, Carlos Bastidas-Caldes

Background

Leishmania is the parasitic protozoan responsible for leishmaniases, a disease that can cause a range of cutaneous, mucosal, and visceral infections. Two subgenera L. Viannia and L. Leishmania are known to infect humans in the tropics and subtropics of the Americas. The

Conclusion

The new primers targeting different two genes and standardized in a PCR assay could identified and differentiated Leishmania parasites at subgenus level. This protocol could be used for Leishmania genus identification and diagnosis at the subgenus level and for determining the parasite's geographical distribution where different Leishmania subgenera are found in the same area.

Methods

We designed two new pairs of primers for a PCR method from two conserved genes, cysteine proteinase B (cpb) and N-acetylglucosamine-6-phosfate deacetylase-like protein (nagA), as specific markers for those two respective subgenera. Primers were tested with 16 microscopical positive clinical samples from the Amazon region of Ecuador obtained in 2010-2020 period.

Results

The cpb presented a band of 172 bp and the nagA a band of 300 bp, thus clearly differentiating L. viannia from L. leishmania. Additionally, primers identified and differentiated the clinical samples in the two subgenera.

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