Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and Brazil

Abstract Background Visceral leishmaniasis (VL) an ‘infectious disease of poverty’, caused by the Leishmania donovani complex, remains a significant public health threat in endemic regions of South Asia, East Africa, and Brazil. Early and accurate diagnosis is critical to prevent the disease's...

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Main Authors: Anirban Bhattacharyya, Nicky Didwania, Sarfaraz Ahmad Ejazi, Rudra Chhajer, Saswati Gayen, Mehebubar Rahman, Rama Prosad Goswami, Krishna Pandey, Vidya Nand Ravi Das, Pradeep Das, Fernando Oliveira da Silva, Dorcas Lamounier Costa, Carlos Henrique Nery Costa, Nahid Ali
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Infectious Diseases of Poverty
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Online Access:https://doi.org/10.1186/s40249-025-01296-7
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author Anirban Bhattacharyya
Nicky Didwania
Sarfaraz Ahmad Ejazi
Rudra Chhajer
Saswati Gayen
Mehebubar Rahman
Rama Prosad Goswami
Krishna Pandey
Vidya Nand Ravi Das
Pradeep Das
Fernando Oliveira da Silva
Dorcas Lamounier Costa
Carlos Henrique Nery Costa
Nahid Ali
author_facet Anirban Bhattacharyya
Nicky Didwania
Sarfaraz Ahmad Ejazi
Rudra Chhajer
Saswati Gayen
Mehebubar Rahman
Rama Prosad Goswami
Krishna Pandey
Vidya Nand Ravi Das
Pradeep Das
Fernando Oliveira da Silva
Dorcas Lamounier Costa
Carlos Henrique Nery Costa
Nahid Ali
author_sort Anirban Bhattacharyya
collection DOAJ
description Abstract Background Visceral leishmaniasis (VL) an ‘infectious disease of poverty’, caused by the Leishmania donovani complex, remains a significant public health threat in endemic regions of South Asia, East Africa, and Brazil. Early and accurate diagnosis is critical to prevent the disease's potentially fatal outcomes. However, due to the nonspecific nature of clinical symptoms, diagnosis often relies on serological tests. This study aims to assess the diagnostic potential of the L. donovani activated C kinase (LACK), a highly conserved antigen essential for parasite survival and host establishment, in VL-endemic regions such as India and Brazil. Methods We conducted a multi-center study with serum samples from India (n = 184) and Brazil (n = 59), along with non-invasive urine samples from India (n = 132). Clinical samples from India were collected from the endemic regions of Bihar and West Bengal between 2016–2024, while those from Teresina, Brazil, were collected between 2008 and 2009. Following preliminary immunoblot analysis, we validated the diagnostic utility of LACK through enzyme-linked immunosorbent assays (ELISA) and dipstick tests. Results were analyzed and area under a Receiver Operating Characteristic (ROC) curve (AUC) values were calculated via the Mann–Whitney U test. Additionally, sensitivity, specificity, and confidence intervals were assessed to evaluate diagnostic performance. Results The ELISA results revealed that LACK antibodies exhibited 100% sensitivity in both Indian [95% confidence intervals (CI): 94.80–100%] and Brazilian (95% CI: 91.24–100%) patient samples, with specificity of 97.33% for Indian controls and 94.74% for Brazilian controls. Urine samples from Indian patients also demonstrated perfect sensitivity and specificity (100%). Notably, LACK showed minimal reactivity with follow-up patient samples. Dipstick assays confirmed these findings, offering a simple, rapid, and field-friendly diagnostic alternative. Conclusion LACK is a promising diagnostic marker for VL, showing high sensitivity across regions and has potential to distinguish active infections from cured or relapsed cases, though larger studies are needed for confirmation. Graphical Abstract
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spelling doaj-art-bf2eda8d97574edda9745f4293382dca2025-08-20T04:02:41ZengBMCInfectious Diseases of Poverty2049-99572025-07-0114111410.1186/s40249-025-01296-7Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and BrazilAnirban Bhattacharyya0Nicky Didwania1Sarfaraz Ahmad Ejazi2Rudra Chhajer3Saswati Gayen4Mehebubar Rahman5Rama Prosad Goswami6Krishna Pandey7Vidya Nand Ravi Das8Pradeep Das9Fernando Oliveira da Silva10Dorcas Lamounier Costa11Carlos Henrique Nery Costa12Nahid Ali13(CSIR)-Indian Institute of Chemical Biology, Infectious Diseases and Immunology Division(CSIR)-Indian Institute of Chemical Biology, Infectious Diseases and Immunology Division(CSIR)-Indian Institute of Chemical Biology, Infectious Diseases and Immunology Division(CSIR)-Indian Institute of Chemical Biology, Infectious Diseases and Immunology DivisionDepartment of Microbiology, Vijaygarh Jyotish Ray CollegeDepartment of Tropical Medicine, School of Tropical MedicineDepartment of Tropical Medicine, School of Tropical MedicineDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical SciencesDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical SciencesDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical SciencesThe Federal University of Piauí (UFPI)The Federal University of Piauí (UFPI)The Federal University of Piauí (UFPI)(CSIR)-Indian Institute of Chemical Biology, Infectious Diseases and Immunology DivisionAbstract Background Visceral leishmaniasis (VL) an ‘infectious disease of poverty’, caused by the Leishmania donovani complex, remains a significant public health threat in endemic regions of South Asia, East Africa, and Brazil. Early and accurate diagnosis is critical to prevent the disease's potentially fatal outcomes. However, due to the nonspecific nature of clinical symptoms, diagnosis often relies on serological tests. This study aims to assess the diagnostic potential of the L. donovani activated C kinase (LACK), a highly conserved antigen essential for parasite survival and host establishment, in VL-endemic regions such as India and Brazil. Methods We conducted a multi-center study with serum samples from India (n = 184) and Brazil (n = 59), along with non-invasive urine samples from India (n = 132). Clinical samples from India were collected from the endemic regions of Bihar and West Bengal between 2016–2024, while those from Teresina, Brazil, were collected between 2008 and 2009. Following preliminary immunoblot analysis, we validated the diagnostic utility of LACK through enzyme-linked immunosorbent assays (ELISA) and dipstick tests. Results were analyzed and area under a Receiver Operating Characteristic (ROC) curve (AUC) values were calculated via the Mann–Whitney U test. Additionally, sensitivity, specificity, and confidence intervals were assessed to evaluate diagnostic performance. Results The ELISA results revealed that LACK antibodies exhibited 100% sensitivity in both Indian [95% confidence intervals (CI): 94.80–100%] and Brazilian (95% CI: 91.24–100%) patient samples, with specificity of 97.33% for Indian controls and 94.74% for Brazilian controls. Urine samples from Indian patients also demonstrated perfect sensitivity and specificity (100%). Notably, LACK showed minimal reactivity with follow-up patient samples. Dipstick assays confirmed these findings, offering a simple, rapid, and field-friendly diagnostic alternative. Conclusion LACK is a promising diagnostic marker for VL, showing high sensitivity across regions and has potential to distinguish active infections from cured or relapsed cases, though larger studies are needed for confirmation. Graphical Abstracthttps://doi.org/10.1186/s40249-025-01296-7L. donovani activated C kinase (LACK)Visceral leishmaniasisDiagnosisELISADipstick
spellingShingle Anirban Bhattacharyya
Nicky Didwania
Sarfaraz Ahmad Ejazi
Rudra Chhajer
Saswati Gayen
Mehebubar Rahman
Rama Prosad Goswami
Krishna Pandey
Vidya Nand Ravi Das
Pradeep Das
Fernando Oliveira da Silva
Dorcas Lamounier Costa
Carlos Henrique Nery Costa
Nahid Ali
Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and Brazil
Infectious Diseases of Poverty
L. donovani activated C kinase (LACK)
Visceral leishmaniasis
Diagnosis
ELISA
Dipstick
title Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and Brazil
title_full Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and Brazil
title_fullStr Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and Brazil
title_full_unstemmed Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and Brazil
title_short Recombinant Leishmania-activated C kinase as a novel antigenic candidate for immuno-diagnosis of visceral leishmaniasis occurring in India and Brazil
title_sort recombinant leishmania activated c kinase as a novel antigenic candidate for immuno diagnosis of visceral leishmaniasis occurring in india and brazil
topic L. donovani activated C kinase (LACK)
Visceral leishmaniasis
Diagnosis
ELISA
Dipstick
url https://doi.org/10.1186/s40249-025-01296-7
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