Isolation and characterisation of Leishmania donovani protein antigens from urine of visceral leishmaniasis patients

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. don...

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Bibliographic Details
Published in:PloS one 2020-09, Vol.15 (9), p.e0238840-e0238840
Main Authors: Marlais, Tegwen, Bhattacharyya, Tapan, Pearson, Callum, Gardner, Bathsheba L, Marhoon, Safiyyah, Airs, Stephanie, Hayes, Kiera, Falconar, Andrew K, Singh, Om Prakash, Reed, Steven G, El-Safi, Sayda, Sundar, Shyam, Miles, Michael A
Format: Article
Language:English
Subjects:
Antibodies
Antigens
Biology
Biology and Life Sciences
Diagnostic systems
Epitopes
Ethics
Health aspects
Health care
Hygiene
Identification and classification
Immunoassays
Infections
Kinases
Leishmania
Leishmania donovani
Mass spectrometry
Mass spectroscopy
Medicine
Medicine and Health Sciences
Methods
Parasites
Parasitic diseases
Patients
Physiological aspects
Polyclonal antibodies
Protein research
Proteins
Research and Analysis Methods
Ribosomal protein S9
Scientific imaging
Serology
Supervision
Tropical diseases
Urinalysis
Urine
Vector-borne diseases
Visceral leishmaniasis
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0238840