Advanced search
Start date
Betweenand


Parasite Detection in Visceral Leishmaniasis Samples by Dye-Based qPCR Using New Gene Targets of Leishmania infantum and Crithidia

Full text
Author(s):
Show less -
Takamiya, Nayore Tamie ; Rogerio, Luana Aparecida ; Torres, Caroline ; Leonel, Joao Augusto Franco ; Vioti, Geovanna ; Oliveira, Tricia Maria Ferreira de Sousa ; Valeriano, Karoline Camila ; Porcino, Gabriane Nascimento ; Santos, Isabel Kinney Ferreira de Miranda ; Costa, Carlos H. N. ; Costa, Dorcas Lamounier ; Ferreira, Tauana Sousa ; Gurgel-Goncalves, Rodrigo ; da Silva, Joao Santana ; Teixeira, Felipe Roberti ; De Almeida, Roque Pacheco ; Ribeiro, Jose M. C. ; Maruyama, Sandra Regina
Total Authors: 18
Document type: Journal article
Source: TROPICAL MEDICINE AND INFECTIOUS DISEASE; v. 8, n. 8, p. 25-pg., 2023-08-01.
Abstract

Visceral leishmaniasis (VL) is a neglected disease considered a serious public health problem, especially in endemic countries. Several studies have discovered monoxenous trypanosomatids (Leptomonas and Crithidia) in patients with VL. In different situations of leishmaniasis, investigations have examined cases of co-infection between Leishmania spp. and Crithidia spp. These coinfections have been observed in a wide range of vertebrate hosts, indicating that they are not rare. Diagnostic techniques require improvements and more robust tools to accurately detect the causative agent of VL. This study aimed to develop a real-time quantitative dye-based PCR (qPCR) assay capable of distinguishing Leishmania infantum from Crithidia-related species and to estimate the parasite load in samples of VL from humans and animals. The primer LinJ31_2420 targets an exclusive phosphatase of L. infantum; the primer Catalase_LVH60-12060_1F targets the catalase gene of Crithidia. Therefore, primers were designed to detect L. infantum and Crithidia sp. LVH60A (a novel trypanosomatid isolated from VL patients in Brazil), in samples related to VL. These primers were considered species-specific, based on sequence analysis using genome data retrieved from the TriTryp database and the genome assembling of Crithidia sp. LVH60A strain, in addition to experimental and clinical data presented herein. This novel qPCR assay was highly accurate in identifying and quantifying L. infantum and Crithidia sp. LVH60A in samples obtained experimentally (in vitro and in vivo) or collected from hosts (humans, dogs, cats, and vectors). Importantly, the screening of 62 cultured isolates from VL patients using these primers surprisingly revealed that 51 parasite cultures were PCR+ for Crithidia sp. In addition, qPCR assays identified the co-infection of L. infantum with Crithidia sp. LVH60A in two new VL cases in Brazil, confirming the suspicion of co-infection in a previously reported case of fatal VL. We believe that the species-specific genes targeted in this study can be helpful for the molecular diagnosis of VL, as well as for elucidating suspected co-infections with monoxenous-like trypanosomatids, which is a neglected fact of a neglected disease. (AU)

FAPESP's process: 21/12464-8 - Developing molecular tools for the study of emerging parasites in Visceral Leishmaniasis
Grantee:Nayore Tamie Takamiya
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 16/18527-3 - The role of IgG FC N-glycosylation on the pathogenesis of visceral leishmaniasis: insights into new strategies of therapy
Grantee:Gabriane Nascimento Porcino
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 18/26799-9 - Phenotypic characterization and genomic analysis of Crithidia-like parasites obtained from patients diagnosed with visceral leishmaniasis
Grantee:Luana Aparecida Rogerio
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 19/19789-0 - Exploratory studies on factors that affect outcomes of infections with Leishmania infantum in humans
Grantee:Isabel Kinney Ferreira de Miranda Santos
Support Opportunities: Regular Research Grants
FAPESP's process: 16/20258-0 - Visceral leishmaniasis: genomics approaches for integrated molecular analysis of host and parasite
Grantee:Sandra Regina Costa Maruyama
Support Opportunities: Research Grants - Young Investigators Grants
FAPESP's process: 21/10358-6 - Genomic analysis of Leishmaniinae parasites isolated from human visceral Leishmaniasis
Grantee:Caroline Torres
Support Opportunities: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 20/15771-6 - Functional characterization of the CRLs (Cullin RING ligases) E3 ubiquitin-ligases in Leishmania infantum
Grantee:Felipe Roberti Teixeira
Support Opportunities: Regular Research Grants
FAPESP's process: 20/14011-8 - Phenotypic characterization and genomic analysis of Crithidia-like parasites obtained from patients diagnosed with Visceral Leishmaniasis
Grantee:Luana Aparecida Rogerio
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 17/16328-6 - Visceral Leishmaniasis: genomics approaches for integrated molecular analysis of host and parasite.
Grantee:Sandra Regina Costa Maruyama
Support Opportunities: Scholarships in Brazil - Young Researchers