Development of a sandwich ELISA to detect Leishmania 40S ribosomal protein S12 antigen from blood samples of visceral leishmaniasis patients

Autor: Greg Matlashewski, Pradeep Das, Raodoh Mohamath, Ayan Kumar Ghosh, Randall F. Howard, Wen-Wei Zhang, Alessandro Picone, Momar Ndao, Steven G. Reed, Patrick Lypaczewski, Jacqueline Whittle
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Male
Parasitemia
40S ribosomal protein S12
Parasite load
Parasite Load
0302 clinical medicine
Diagnosis
Asymptomatic Infections
Leishmaniasis
Leishmania
Visceral Leishmaniasis
biology
Neglected Diseases
Middle Aged
3. Good health
Infectious Diseases
Leishmaniasis
Visceral
Female
ELISA
Rabbits
Antibody
Research Article
Adult
Ribosomal Proteins
Adolescent
030231 tropical medicine
Antigens
Protozoan
Enzyme-Linked Immunosorbent Assay
Sensitivity and Specificity
lcsh:Infectious and parasitic diseases
03 medical and health sciences
Antigen
parasitic diseases
medicine
Animals
Humans
lcsh:RC109-216
Post-kala-azar dermal leishmaniasis
business.industry
PBMC
medicine.disease
biology.organism_classification
Virology
030104 developmental biology
Visceral leishmaniasis
Case-Control Studies
biology.protein
Leukocytes
Mononuclear
business
Zdroj: BMC Infectious Diseases, Vol 18, Iss 1, Pp 1-11 (2018)
BMC Infectious Diseases
ISSN: 1471-2334
DOI: 10.1186/s12879-018-3420-2
Popis: Background Visceral leishmaniasis (VL), caused by Leishmania donovani complex parasites, is a neglected parasitic disease that is generally fatal if untreated. Despite decades of research to develop a sensitive VL diagnostic test, definitive diagnosis of VL still mainly relies on the visualization of the parasite in aspirates from the spleen, liver or bone marrow, an invasive and dangerous process with variable sensitivity. A sensitive assay that can detect Leishmania antigen from blood samples will help confirm cause, cure or recurrence of VL. Methods In this study, rabbit polyclonal antibodies were raised against eight recombinant Leishmania proteins that are highly abundant in Leishmania. The antibodies were purified and labeled with biotin for developing a prototype sandwich enzyme-linked immunosorbent assay (ELISA). Results The ELISA for the Leishmania 40S ribosomal protein S12 detected target antigen with the highest sensitivity and specificity and could detect 1 pg of purified protein or as few as 60 L. donovani parasites. The 40S ribosomal protein S12 sandwich ELISA could detect the target antigen from Peripheral Blood Mononuclear Cell (PBMC) samples in 68% of VL patients and post-kala-azar dermal leishmaniasis (PKDL) patients, providing an estimation of parasitemia ranging from 15 to 80 amastigotes per ml of blood. Conclusion These results indicate that the 40S ribosomal protein S12 sandwich ELISA warrants further tests with more clinical samples of VL patients and other parasitic diseases. It is hopeful that this ELISA could become a useful tool for confirming VL diagnosis, monitoring treatment progress, disease recurrence and possibly detecting asymptomatic Leishmania infections with a high parasite load.
Databáze: OpenAIRE
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