Prevalidation of an ELISA for Detection of a New Clinical Entity: Leishmania donovani-Induced Cutaneous Leishmaniasis
Joint Authors
Deepachandi, Bhagya
Andrahennadi, Thisira P.
Wickramanayake, Mahendra N.
Siri, Shantha
Chandrasekharan, Vishvanath
Siriwardana, Yamuna
Weerasinghe, Sudath
Soysa, Preethi
Gunathilake, Himali
Source
International Journal of Analytical Chemistry
Issue
Vol. 2020, Issue 2020 (31 Dec. 2020), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2020-07-15
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Abstract EN
Human leishmaniasis which is considered a neglected tropical parasitic disease presents in three main clinical forms (i.e., cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (MCL), and visceral leishmaniasis (VL)) that are mainly determined by its causative species.
Leishmania donovani, the most virulent and visceralizing parasite, is increasingly reported to cause CL in many countries in the world.
Although CL is generally not considered to evoke a humoral immune response except for a nonrobust and a variable response in minority of cases, VL is associated with a clear strong humoral response.
However, humoral response in L.
donovani-induced CL has not been well evaluated before.
A suitable serology-based assay is an essential primary step in such a study.
An indirect enzyme-linked immunosorbent assay (ELISA) based on Leishmania promastigote crude antigen (Ag) was designed and optimized in order to utilize in further serological studies on this new clinical entity.
Optimization included quantification of crude Ag, checkerboard titration method for determination of optimal concentrations for coating Ag, human sera and secondary antibody (Ab) with suitable coating buffer, blocking buffer, and incubating temperatures.
The selected coating buffer was 0.02 M phosphate buffer, pH 6.8, and the blocking buffer was 2% fetal bovine serum with 0.01 M phosphate-buffered saline.
At least 1 μg of crude Ag was required for coating the ELISA plate, while 1 : 1000 serum was used as primary Ab.
The optimized concentration of secondary Ab was 1 : 64000 which might be altered according to manufacturer recommendations.
The assay specificity was pre-evaluated using sera (n = 20 from each category) from confirmed CL patients and controls (other skin diseases which mimic CL, other systemic diseases that mimic VL, nonendemic healthy controls, and endemic healthy controls).
This procedure described an optimization procedure of an ELISA technique for detection of anti-Leishmania antibodies in patients with L.
donovani caused CL.
American Psychological Association (APA)
Deepachandi, Bhagya& Weerasinghe, Sudath& Gunathilake, Himali& Andrahennadi, Thisira P.& Wickramanayake, Mahendra N.& Siri, Shantha…[et al.]. 2020. Prevalidation of an ELISA for Detection of a New Clinical Entity: Leishmania donovani-Induced Cutaneous Leishmaniasis. International Journal of Analytical Chemistry،Vol. 2020, no. 2020, pp.1-8.
https://search.emarefa.net/detail/BIM-1167816
Modern Language Association (MLA)
Deepachandi, Bhagya…[et al.]. Prevalidation of an ELISA for Detection of a New Clinical Entity: Leishmania donovani-Induced Cutaneous Leishmaniasis. International Journal of Analytical Chemistry No. 2020 (2020), pp.1-8.
https://search.emarefa.net/detail/BIM-1167816
American Medical Association (AMA)
Deepachandi, Bhagya& Weerasinghe, Sudath& Gunathilake, Himali& Andrahennadi, Thisira P.& Wickramanayake, Mahendra N.& Siri, Shantha…[et al.]. Prevalidation of an ELISA for Detection of a New Clinical Entity: Leishmania donovani-Induced Cutaneous Leishmaniasis. International Journal of Analytical Chemistry. 2020. Vol. 2020, no. 2020, pp.1-8.
https://search.emarefa.net/detail/BIM-1167816
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1167816