Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

• Published in: Revista brasileira de parasitologia veterinaria Vol. 21; no. 3; pp. 304 - 307
• Main Authors: da Silveira Neto, Osvaldo José, Duarte, Sabrina Castilho, da Costa, Hérika Xavier, Linhares, Guido Fontgalland Coelho
• Format: Journal Article
• Language: English
• Published: Brazil Colégio Brasileiro de Parasitologia Veterinária 01-09-2012
• Subjects: DNA Primers; Leishmania infantum - genetics; Leishmaniasis, Visceral - diagnosis; PARASITOLOGY; Polymerase Chain Reaction; Species Specificity; VETERINARY SCIENCES; Kala-azar; diagnóstico molecular; leishmaniose visceral; L. infantum chagasi; gene 18S rRNA; 18S rRNA gene; visceral leishmaniasis; Calazar; molecular diagnosis
• ISSN: 1984-2961; 1984-2961
• DOI: 10.1590/s1984-29612012000300024

The objective of this study was to design and evaluate new primers for species-specific detection of L. infantum chagasi using PCR. Two combinations of primer pairs were established with the aim of obtaining specific amplification products from the L. infantum chagasi 18S rRNA gene. The combinations of the primer pairs and the respective sizes of the PCR products, based on the U422465 GenBank reference sequence of L. infantum chagasi, were: LCS1/LCS3 (259 bp) and LCS2/LCS3 (820 bp). It was concluded that the new PCR assays optimized using the primer pairs LCS1/LCS3 and LCS2/LCS3 were effective for specific detection of L. infantum chagasi, with analytical sensitivity to detect 1 pg/µL of DNA.