How to Detect Antibodies Against Babesia divergens in Human Blood Samples
(2024) In Open Forum Infectious Diseases 11(2).- Abstract
Background. Today only indirect fluorescent antibody assays (IFAs) are commercially available to detect antibodies against Babesia divergens in humans. IFA is subjective and requires highly experienced staff. We have therefore developed an enzyme-linked immunosorbent assay (ELISA)-based method for measuring anti-B. divergens immunoglobulin G antibodies in human blood samples. Methods. Crude merozoite extract from in vitro cultures of a new B. divergens isolate was used in ELISA to detect antibodies in different sets of samples: Borrelia burgdorferi-positive samples, healthy individuals, tick-bitten individuals including follow-up samples 3 months later, positive control samples from patients with an active Babesia infection, and samples... (More)
Background. Today only indirect fluorescent antibody assays (IFAs) are commercially available to detect antibodies against Babesia divergens in humans. IFA is subjective and requires highly experienced staff. We have therefore developed an enzyme-linked immunosorbent assay (ELISA)-based method for measuring anti-B. divergens immunoglobulin G antibodies in human blood samples. Methods. Crude merozoite extract from in vitro cultures of a new B. divergens isolate was used in ELISA to detect antibodies in different sets of samples: Borrelia burgdorferi-positive samples, healthy individuals, tick-bitten individuals including follow-up samples 3 months later, positive control samples from patients with an active Babesia infection, and samples from malaria-endemic regions. As a reference, IFA was used to detect antibodies in the tick-bitten samples. Western blot was used to evaluate reactions against specific bands in extracts with/without parasites. Results. Using IFA as the reference method, the sensitivity and specificity of the ELISA were 86% (12/14) and 100% (52/52). There was a very high correlation (r = -0.84; P = .0004) between IFA dilution factors and ELISA absorbances among the samples classified as positive. Five percent of the B. burgdorferi-positive samples were judged as weakly positive and 5% as strongly positive in our ELISA. Western blot showed that the immunodominant antigens (∼120 kDa) were from merozoites and not from erythrocytes. Conclusions. This ELISA can detect antibodies directed against B. divergens, and it can be a useful and easy assay to handle compared with IFA. The ELISA can also measure high and low levels of antibodies, which could give insight into the recency of a B. divergens infection.
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- author
- Tijani, Muyideen Kolapo LU ; Svensson, Joel LU ; Adlerborn, Paula LU ; Danielsson, Lena LU ; Teleka, Alexandra LU ; Lövmar, Matilda Ljungqvist ; Lindgren, Per Eric ; Forsberg, Pia and Persson, Kristina E.M. LU
- organization
-
- Malaria and Babesia (research group)
- Division of Clinical Chemistry and Pharmacology
- Clinical Chemistry, Malmö (research group)
- Clinical Memory Research (research group)
- MultiPark: Multidisciplinary research focused on Parkinson´s disease
- LUCC: Lund University Cancer Centre
- Protease Inhibitor Research (research group)
- publishing date
- 2024-02-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- antibody, Babesia, blood transfusion, divergens, ELISA
- in
- Open Forum Infectious Diseases
- volume
- 11
- issue
- 2
- article number
- ofae028
- publisher
- Oxford University Press
- external identifiers
-
- pmid:38328497
- scopus:85184619978
- ISSN
- 2328-8957
- DOI
- 10.1093/ofid/ofae028
- language
- English
- LU publication?
- yes
- id
- 9170fcfc-ed84-4729-a30f-4ca1d1ed8ba3
- date added to LUP
- 2024-02-22 15:38:48
- date last changed
- 2024-04-23 16:14:04
@article{9170fcfc-ed84-4729-a30f-4ca1d1ed8ba3, abstract = {{<p>Background. Today only indirect fluorescent antibody assays (IFAs) are commercially available to detect antibodies against Babesia divergens in humans. IFA is subjective and requires highly experienced staff. We have therefore developed an enzyme-linked immunosorbent assay (ELISA)-based method for measuring anti-B. divergens immunoglobulin G antibodies in human blood samples. Methods. Crude merozoite extract from in vitro cultures of a new B. divergens isolate was used in ELISA to detect antibodies in different sets of samples: Borrelia burgdorferi-positive samples, healthy individuals, tick-bitten individuals including follow-up samples 3 months later, positive control samples from patients with an active Babesia infection, and samples from malaria-endemic regions. As a reference, IFA was used to detect antibodies in the tick-bitten samples. Western blot was used to evaluate reactions against specific bands in extracts with/without parasites. Results. Using IFA as the reference method, the sensitivity and specificity of the ELISA were 86% (12/14) and 100% (52/52). There was a very high correlation (r = -0.84; P = .0004) between IFA dilution factors and ELISA absorbances among the samples classified as positive. Five percent of the B. burgdorferi-positive samples were judged as weakly positive and 5% as strongly positive in our ELISA. Western blot showed that the immunodominant antigens (∼120 kDa) were from merozoites and not from erythrocytes. Conclusions. This ELISA can detect antibodies directed against B. divergens, and it can be a useful and easy assay to handle compared with IFA. The ELISA can also measure high and low levels of antibodies, which could give insight into the recency of a B. divergens infection.</p>}}, author = {{Tijani, Muyideen Kolapo and Svensson, Joel and Adlerborn, Paula and Danielsson, Lena and Teleka, Alexandra and Lövmar, Matilda Ljungqvist and Lindgren, Per Eric and Forsberg, Pia and Persson, Kristina E.M.}}, issn = {{2328-8957}}, keywords = {{antibody; Babesia; blood transfusion; divergens; ELISA}}, language = {{eng}}, month = {{02}}, number = {{2}}, publisher = {{Oxford University Press}}, series = {{Open Forum Infectious Diseases}}, title = {{How to Detect Antibodies Against Babesia divergens in Human Blood Samples}}, url = {{http://dx.doi.org/10.1093/ofid/ofae028}}, doi = {{10.1093/ofid/ofae028}}, volume = {{11}}, year = {{2024}}, }