The use of kDNA minicircle subclass relative abundance to differentiate between Leishmania (L.) infantum and Leishmania (L.) amazonensis

Parasites & Vectors
Marcello CeccarelliMauro Magnani

Abstract

Leishmaniasis is a neglected disease caused by many Leishmania species, belonging to subgenera Leishmania (Leishmania) and Leishmania (Viannia). Several qPCR-based molecular diagnostic approaches have been reported for detection and quantification of Leishmania species. Many of these approaches use the kinetoplast DNA (kDNA) minicircles as the target sequence. These assays had potential cross-species amplification, due to sequence similarity between Leishmania species. Previous works demonstrated discrimination between L. (Leishmania) and L. (Viannia) by SYBR green-based qPCR assays designed on kDNA, followed by melting or high-resolution melt (HRM) analysis. Importantly, these approaches cannot fully distinguish L. (L.) infantum from L. (L.) amazonensis, which can coexist in the same geographical area. DNA from 18 strains/isolates of L. (L.) infantum, L. (L.) amazonensis, L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, and 62 clinical samples from L. (L.) infantum-infected dogs were amplified by a previously developed qPCR (qPCR-ML) and subjected to HRM analysis; selected PCR products were sequenced using an ABI PRISM 310 Genetic Analyzer. Based on the obtained sequences, a new SYBR-green qPCR assay (qPCR-ama) in...Continue Reading

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Citations

Sep 20, 2019·Transboundary and Emerging Diseases·Taiane Acunha EscobarVanusa Manfredini
Jul 16, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Susiany P LopesDamião P de Sousa
May 3, 2018·Parasites & Vectors·Luca GalluzziMauro Magnani
Jun 21, 2018·Molecular Diagnosis & Therapy·Shyam Sundar, Om Prakash Singh
Jun 6, 2018·Frontiers in Microbiology·Aurora DiotalleviMauro Magnani
Mar 18, 2021·Revista brasileira de parasitologia veterinária = Brazilian journal of veterinary parasitology : Órgão Oficial do Colégio Brasileiro de Parasitologia Veterinária·Beatriz Batista TrigoCáris Maroni Nunes
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May 25, 2021·Veterinary Parasitology : Regional Studies and Reports·Ayça AksuluSerkan Bakırcı
Oct 15, 2020·Experimental Parasitology·Rayana Carla Silva de MoraisMilena de Paiva Cavalcanti

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Datasets Mentioned

BETA
M21325

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

CLUSTAL
Gene 6000
BLAST
Rotor
MUSCLE
MEGA
GraphPad
qPCR
GraphPad InStat
ML

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