Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India
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, sp...
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MDPI AG
2022-01-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/11/2/120 |
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| author | Anirban Bhattacharyya Mohd Kamran Sarfaraz Ahmad Ejazi Sonali Das Nicky Didwania Rahul Bhattacharjee Mehebubar Rahaman Rama Prosad Goswami Krishna Pandey Vidya Nand Ravi Das Pradeep Das Saswati Gayen Nahid Ali |
| author_facet | Anirban Bhattacharyya Mohd Kamran Sarfaraz Ahmad Ejazi Sonali Das Nicky Didwania Rahul Bhattacharjee Mehebubar Rahaman Rama Prosad Goswami Krishna Pandey Vidya Nand Ravi Das Pradeep Das Saswati Gayen Nahid Ali |
| author_sort | Anirban Bhattacharyya |
| collection | DOAJ |
| description | 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 <i>Leishmania</i> 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. |
| first_indexed | 2024-03-09T21:16:50Z |
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| id | doaj.art-518fc8ba74984a149b8aa7b2a073df8e |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-09T21:16:50Z |
| publishDate | 2022-01-01 |
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| spelling | doaj.art-518fc8ba74984a149b8aa7b2a073df8e2023-11-23T21:30:41ZengMDPI AGPathogens2076-08172022-01-0111212010.3390/pathogens11020120Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in IndiaAnirban Bhattacharyya0Mohd Kamran1Sarfaraz Ahmad Ejazi2Sonali Das3Nicky Didwania4Rahul Bhattacharjee5Mehebubar Rahaman6Rama Prosad Goswami7Krishna Pandey8Vidya Nand Ravi Das9Pradeep Das10Saswati Gayen11Nahid Ali12Infectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata 700032, IndiaInfectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata 700032, IndiaInfectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata 700032, IndiaInfectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata 700032, IndiaInfectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata 700032, IndiaInfectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata 700032, IndiaDepartment of Tropical Medicine, School of Tropical Medicine, Kolkata 700073, IndiaDepartment of Tropical Medicine, School of Tropical Medicine, Kolkata 700073, IndiaDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, IndiaDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, IndiaDepartment of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, IndiaDepartment of Microbiology, VijaygarhJyotish Ray College, Bejoygarh 700032, IndiaInfectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata 700032, IndiaVisceral 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 <i>Leishmania</i> 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.https://www.mdpi.com/2076-0817/11/2/120LeishmaniasisdiagnosisantigenELISAimmunoblotting |
| spellingShingle | Anirban Bhattacharyya Mohd Kamran Sarfaraz Ahmad Ejazi Sonali Das Nicky Didwania Rahul Bhattacharjee Mehebubar Rahaman Rama Prosad Goswami Krishna Pandey Vidya Nand Ravi Das Pradeep Das Saswati Gayen Nahid Ali Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India Pathogens Leishmaniasis diagnosis antigen ELISA immunoblotting |
| title | Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India |
| title_full | Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India |
| title_fullStr | Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India |
| title_full_unstemmed | Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India |
| title_short | Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India |
| title_sort | revealing a novel antigen repressor of differentiation kinase 2 for diagnosis of human visceral leishmaniasis in india |
| topic | Leishmaniasis diagnosis antigen ELISA immunoblotting |
| url | https://www.mdpi.com/2076-0817/11/2/120 |
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