Browsing by Author "Callum Pearson"

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Isolation and characterisation of Leishmania donovani protein antigens from urine of visceral leishmaniasis patients
(Public Library of Science, 2020) Tegwen Marlais; Tapan Bhattacharyya; Callum Pearson; Bathsheba L. Gardner; Safiyyah Marhoon; Stephanie Airs; Kiera Hayes; Andrew K. Falconar; Om Prakash Singh; Steven G. Reed; Sayda El-Safi; Shyam Sundar; Michael A. Miles
Diagnosis of visceral leishmaniasis (VL) relies on invasive and risky aspirate procedures, and confirmation of cure after treatment is unreliable. Detection of Leishmania donovani antigens in urine has the potential to provide both a non-invasive diagnostic and a test of cure. We searched for L. donovani antigens in urine of VL patients from India and Sudan to contribute to the development of urine antigen capture immunoassays. VL urine samples were incubated with immobilised anti-L. donovani polyclonal antibodies and captured material was eluted. Sudanese eluted material and concentrated VL urine were analysed by western blot. Immunocaptured and immunoreactive material from Indian and Sudanese urine was submitted to mass spectrometry for protein identification. We identified six L. donovani proteins from VL urine. Named proteins were 40S ribosomal protein S9, kinases, and others were hypothetical. Thirty-three epitope regions were predicted with high specificity in the 6 proteins. Of these, 20 were highly specific to Leishmania spp. and are highly suitable for raising antibodies for the subsequent development of an antigen capture assay. We present all the identified proteins and analysed epitope regions in full so that they may contribute to the development of non-invasive immunoassays for this deadly disease. © 2020 Marlais et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Visceral Leishmaniasis IgG1 Rapid Monitoring of Cure vs. Relapse, and Potential for Diagnosis of Post Kala-Azar Dermal Leishmaniasis
(Frontiers Media S.A., 2018) Tegwen Marlais; Tapan Bhattacharyya; Om Prakash Singh; Pascal Mertens; Quentin Gilleman; Caroline Thunissen; Bruno C. Bremer Hinckel; Callum Pearson; Bathsheba L. Gardner; Stephanie Airs; Marianne de la Roche; Kiera Hayes; Hannah Hafezi; Andrew K. Falconar; Osama Eisa; Alfarazdeg Saad; Basudha Khanal; Narayan Raj Bhattarai; Suman Rijal; Marleen Boelaert; Sayda El-Safi; Shyam Sundar; Michael A. Miles
Background: There is a recognized need for an improved diagnostic test to assess post-chemotherapeutic treatment outcome in visceral leishmaniasis (VL) and to diagnose post kala-azar dermal leishmaniasis (PKDL). We previously demonstrated by ELISA and a prototype novel rapid diagnostic test (RDT), that high anti-Leishmania IgG1 is associated with post-treatment relapse versus cure in VL. Methodology: Here, we further evaluate this novel, low-cost RDT, named VL Sero K-SeT, and ELISA for monitoring IgG1 levels in VL patients after treatment. IgG1 levels against L. donovani lysate were determined. We applied these assays to Indian sera from cured VL at 6 months post treatment as well as to relapse and PKDL patients. Sudanese sera from pre- and post-treatment and relapse were also tested. Results: Of 104 paired Indian sera taken before and after treatment for VL, when deemed clinically cured, 81 (77.9%) were positive by VL Sero K-SeT before treatment; by 6 months, 68 of these 81 (84.0%) had a negative or reduced RDT test line intensity. ELISAs differed in positivity rate between pre- and post-treatment (p = 0.0162). Twenty eight of 33 (84.8%) Indian samples taken at diagnosis of relapse were RDT positive. A comparison of Indian VL Sero K-SeT data from patients deemed cured and relapsed confirmed that there was a significant difference (p < 0.0001) in positivity rate for the two groups using this RDT. Ten of 17 (58.8%) Sudanese sera went from positive to negative or decreased VL Sero K-SeT at the end of 11–30 days of treatment. Forty nine of 63 (77.8%) PKDL samples from India were positive by VL Sero K-SeT. Conclusion: We have further shown the relevance of IgG1 in determining clinical status in VL patients. A positive VL Sero K-SeT may also be helpful in supporting diagnosis of PKDL. With further refinement, such as the use of specific antigens, the VL Sero K-SeT and/or IgG1 ELISA may be adjuncts to current VL control programmes. © Copyright © 2018 Marlais, Bhattacharyya, Singh, Mertens, Gilleman, Thunissen, Hinckel, Pearson, Gardner, Airs, de la Roche, Hayes, Hafezi, Falconar, Eisa, Saad, Khanal, Bhattarai, Rijal, Boelaert, El-Safi, Sundar and Miles.