Evaluation of Leishmania infantum pyridoxal kinase protein for the diagnosis of human and canine visceral leishmaniasis

Oliveira‐da‐Silva, João A.; Machado, Amanda S.; Ramos, Fernanda F.; Tavares, Grasiele S.V.; Lage, Daniela P.; Ludolf, Fernanda; Steiner, Bethina T.; Reis, Thiago A.R.; Santos, Thaís T.O.; Costa, Lourena E.; Martins, Vívian T.; Galvani, Nathália Coral; Chaves, Ana Cristina; Oliveira, Jamil S.; Chávez-Fumagalli, Miguel Á.; Magalhães-Soares, Danielle F. de; Duarte, Mariana C.; Menezes‐Souza, Daniel; Silveira, Júlia A.G.; Moreira, Ricardo Luiz Fontes; Machado-de-Ávila, Ricardo Andrez; Tupinambás, Unaí; Gonçalves, Denise Utsch; Coelho, Eduardo Antônio Ferraz

doi:10.1016/j.imlet.2020.01.005

Cited by 9 publications

(11 citation statements)

References 58 publications

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“…[ 82 ], in which the cases were selected from a health service or hospital. To assess the performance of peptide-based tests, three different formats were reported: one study used an ICT format [ 7 ], two studies used phage-ELISA [ 15 , 62 ] and 19 studies used an ELISA technique [ 10 , 11 , 16 , 17 , 26 , 27 , 29 , 35 , 37 , 40 – 43 , 47 , 48 , 58 , 63 , 69 , 70 ]. In these studies using an ELISA method, a marked difference was observed in the amount of peptides used, which ranged from 0.25 μg to 20 μg per well.…”

Section: Resultsmentioning

confidence: 99%

A systematic review of peptide-based serological tests for the diagnosis of leishmaniasis

et al. 2023

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Serological methods should meet the needs of leishmaniasis diagnosis due to their high sensitivity and specificity, economical and adaptable rapid diagnostic test format, and ease of use. Currently, the performances of serological diagnostic tests, despite improvements with recombinant proteins, vary greatly depending on the clinical form of leishmaniasis and the endemic area. Peptide-based serological tests are promising as they could compensate for antigenic variability and improve performance, independently of Leishmania species and subspecies circulating in the endemic areas. The objective of this systematic review was to inventory all studies published from 2002 to 2022 that evaluate synthetic peptides for serological diagnosis of human leishmaniases and also to highlight the performance (e.g., sensitivity and specificity) of each peptide reported in these studies. All clinical forms of leishmaniasis, visceral and tegumentary, and all Leishmania species responsible for these diseases were considered. Following PRISMA statement recommendations, 1,405 studies were identified but only 22 articles met the selection criteria and were included in this systematic review. These original research articles described 77 different peptides, of which several have promising performance for visceral or tegumentary leishmaniasis diagnosis. This review highlights the importance of and growing interest in synthetic peptides used for serological diagnosis of leishmaniases, and their performances compared to some widely used tests with recombinant proteins.

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Section: Resultsmentioning

confidence: 99%

A systematic review of peptide-based serological tests for the diagnosis of leishmaniasis

et al. 2023

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“…6 Bioinformatics assays showed the comparison between Leishmania infantum PLK and Trypanosoma protein sequence had a low similarity, with identity percentage of 61.04 -66.11%. 7 These data indicate PLK has high variation.…”

Section: Introductionmentioning

confidence: 91%

Cloning and Characterization of Pyridoxal Kinase from Geobacillus sp. H6a

Pasri¹,

Champasri²,

Trongpanich³

2022

J. Pure Appl. Microbiol.

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Pyridoxal kinase encoded by pdxK gene, is the important key enzyme in the salvage pathway of vitamin B6 biosynthesis. The enzyme catalyzes the phosphorylation of the 5′ alcohol groups of free form vitamin B6 into their 5′-phosphate forms that requires metal ion and ATP. Pyridoxal kinase have been reported in many organisms except in the thermophilic bacterium. Therefore, this study aimed to clone, express and characterize pyridoxal kinase of Geobacillus sp. H6a isolated from the hot spring in the North of Thailand. The GhpdxK gene (810 base pairs) was inserted into pET28a(+) plasmids at restriction site of NdeI and BamHI and transformed into E.coli BL21(DE3). The expressed pyridoxal kinase of this bacterium exhibits a homodimer, in which each subunit had a molecular mass of about 32 kDa when examined by SDS-PAGE and gel filtration. The enzyme showed maximal activity at 70°C and at pH 8.0. The expressed enzyme obtained in this study was found to be more active (>50%) in the broad pH range (6.0 – 9.0) than those previously reported. This enzyme prefers Mg2+ and also accepts other cations to the less extent. Under optimal conditions, the expressed enzyme has higher affinity toward PN (20 ± 1.35 µM), while it showed the same affinity to pyridoxal (100 ± 0.76 µM) and pyridoxamine (100 ± 1.21 µM). The Km value for ATP and 4-amino-5-hydroxymethyl-2-methylpyridine were 8.99 ± 1.76 µM and 19 ± 0.85 µM, respectively. With high activity at high temperature and active in the broad pH range, it could be considered as a potential candidate for future application particularly bioconversion of vitamin B6.

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“…In the last few years, protein kinases (PKs) of Leishmania sp., are getting highlighted as important cell signalling molecules that bear the potential of being therapeutic as well as diagnostic markers [9,22]. The recombinant antigen repressor of differentiation kinase 2 (RDK2) is a serine/threonine kinase that has a role in eukaryotic cell-cycle regulation and belongs to the NIMA-related kinases (NEK PK) family.…”

Section: Introductionmentioning

confidence: 99%

“…The recombinant antigen repressor of differentiation kinase 2 (RDK2) is a serine/threonine kinase that has a role in eukaryotic cell-cycle regulation and belongs to the NIMA-related kinases (NEK PK) family. Previously, the roles of RDK2 were studied in controlling the differentiation of Trypanosoma brucei from short stumpy bloodstream form (BSF) to insect procyclic form (PCF) [22]. Besides T. brucei, these mitotic kinases are also highly conserved in Leishmania sp., and they have vital involvement in the regulation of the cell cycle.…”

Section: Introductionmentioning

confidence: 99%

Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India

et al. 2022

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Visceral leishmaniasis (VL) is one of the major global health concerns due to its association with morbidity and mortality. All available diagnostic tools have been, until now, unable to provide a very specific and cost-effective mode of detection for VL globally. Therefore, the design of robust, specific, and commercially translatable diagnostic tests is urgently required. Currently, we are attempting to identify and explore the diagnostic potential of a novel parasite antigen. Repressor of differentiation kinase 2 (RDK2), a serine/threonine kinase, has a versatile role in parasite life cycle progression. However, its role as a diagnostic candidate for VL has not been investigated. Herein, we cloned and over-expressed LdRDK2 and studied the recombinant RDK2 for the diagnosis of human VL using serum and urine samples. In silico analysis predicted that RDK2 is conserved among Leishmania species with the least conservation in humans. RDK2 developed immune-reactive bands with antibodies present in VL patients’ sera, and it demonstrated no cross-reactivity with sera from healthy controls and other diseases. Additionally, RDK2 antigen demonstrated a significant reactivity with IgG antibodies of VL patients’ sera, with 78% sensitivity and 86.67% specificity as compared to healthy controls and other diseases. Furthermore, we evaluated its utility for non-invasive diagnosis of VL using patients’ urine samples and found 93.8% sensitivity and 85.7% specificity. RDK2 was found to have better sensitivity and treatment response in patients’ urine compared to serum samples, indicating its role as a promising point of care (POC) antigen. In a nutshell, we explored the role of RDK2 as a potential diagnostic marker for VL in both invasive and non-invasive modes as well as its utility as a promising POC antigen for treatment response cases.

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Evaluation of Leishmania infantum pyridoxal kinase protein for the diagnosis of human and canine visceral leishmaniasis

Cited by 9 publications

References 58 publications

A systematic review of peptide-based serological tests for the diagnosis of leishmaniasis

A systematic review of peptide-based serological tests for the diagnosis of leishmaniasis

Cloning and Characterization of Pyridoxal Kinase from Geobacillus sp. H6a

Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India

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