Analysis of the IGS RRNA Region and Applicability for (.) Characterization

Overview

Journal J Parasitol Res

Publisher Wiley

Specialty Parasitology

Date 2020 Oct 21

PMID 33083046

Citations 1

Authors

Tayna C de Goes

Rayana C S de Morais

Maria G N de Melo

Romulo P E Silva

Antonio M Rezende

Milena de Paiva-Cavalcanti

Affiliations

Soon will be listed here.

Abstract

The causative species is an important factor influencing the evolution of American cutaneous leishmaniasis (ACL). Due to its wide distribution in endemic areas, (.) is considered one of the most important species in circulation in Brazil. Molecular targets derived from ribosomal RNA (rRNA) were used in studies to identify spp.; however, the Intergenic Spacer (IGS) region has not yet been explored in parasite species differentiation. Besides, there is a shortage of sequences deposited in public repositories for this region. Thus, it was proposed to analyze and provide sequences of the IGS rRNA region from different spp. and to evaluate their potential as biomarkers to characterize . A set of primers was designed for complete amplification of the IGS rRNA region of spp. PCR products were submitted to Sanger sequencing. The sequences obtained were aligned and analyzed for size and similarity, as well as deposited in GenBank. Characteristics of the repetitive elements (IGSRE) present in the IGS rRNA were also verified. In addition, a set of primers for identification for qPCR was developed and optimized. Sensitivity (), specificity (), and efficiency () tests were applied. It was found that the mean size for the IGS rRNA region is 3 kb, and the similarity analysis of the sequences obtained demonstrated high conservation among the species. It was observed that the size for the IGSRE repetitive region varies between 61 and 71 bp, and there is a high identity between some species. Fifteen sequences generated for the IGS rRNA partial region of nine different species were deposited in GenBank so far. The specific primer system for showed = 10 fg, = 98.08%, and log = 10 for ; log = 10 for ; and log = 10 for . This protocol system can be used for diagnosis, identification, and quantification of a patient's parasite load, aiding in the direction of a more appropriate therapeutic management to the cases of infection by this etiological agent. Besides that, the unpublished sequences deposited in databases can be used for multiple analyses in different contexts.

Citing Articles

Laboratory diagnostics for human Leishmania infections: a polymerase chain reaction-focussed review of detection and identification methods.

Gow I, Smith N, Stark D, Ellis J Parasit Vectors. 2022; 15(1):412.

PMID: 36335408 PMC: 9636697. DOI: 10.1186/s13071-022-05524-z.

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