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How to Detect Antibodies Against Babesia divergens in Human Blood Samples

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 (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
; ; ; ; ; ; ; and
organization
publishing date
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}},
}