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Development of an assay for quantification of linkage-specific O-acetylated sialoglycans on erythrocytes; its application in Indian visceral leishmaniasis

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Title Development of an assay for quantification of linkage-specific
O-acetylated sialoglycans on erythrocytes;
its application in Indian visceral leishmaniasis
 
Creator Chava, Anil Kumar
Chatterjee, Mitali
Nandi, Shyam Sundar
Mandal, Chitra
 
Subject Infectious Diseases and Immunology
 
Description We have developed a noninvasive approach for the quantification of linkage-specific 9-O-acetylated sialoglycans on
mammalian erythrocytes using a lectin, Achatinin-H, whose lectinogenic epitope has previously been defined as 9-O-acetylated
sialoglycoconjugates (9-O-AcSGs) a2!6 linked to subterminal GalNAc. Titration and checkerboard analysis were performed
to optimize the assay using rabbit, rat and human erythrocytes that contain differing amounts of this glycotope. Assay
specificity was established by decreased binding of erythrocytes to immobilised Achatinin-H when pre-incubated with excess
lectin. The intra-assay coefficient of variation (CV) for rat and human erythrocytes was 8.6–9.2% and 11.1–13.0%,
respectively. The inter-assay CV for rat and human erythrocytes was 9.9–10.1% and 15.2–16.6%, respectively. In previous
studies, we have identified an enhanced presence of cell surface 9-O-AcSGs on the erythrocytes of patients with visceral
leishmaniasis (VL) [Am. J. Trop. Med. Hyg. 58 (1998) 551]. Our assay when evaluated on erythrocytes from VL patients
(n = 30) showed a fourfold increase in lectin binding as compared to endemic controls. The meanFS.E.M. of the A405 nm value
was 1.14F0.04 vs. 0.23F0.03, respectively ( p < 0.0001). Following effective chemotherapy, a significant reduction of this
glycotope on the erythrocytes of VL patients indicates that this assay has both a diagnostic and prognostic potential. Taken
together, we conclude that this antigen-based assay is a specific and reproducible method for monitoring the disease status of
VL patients and could be used in retrospective and prospective trials.
 
Publisher Elsevier
 
Date 2002
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/769/1/JOURNAL_OF_IMMUNOLOGICAL_METHODS__Volume_270___Issue_1___Pages_1%2D10;2002[9].pdf
Chava, Anil Kumar and Chatterjee, Mitali and Nandi, Shyam Sundar and Mandal, Chitra (2002) Development of an assay for quantification of linkage-specific O-acetylated sialoglycans on erythrocytes; its application in Indian visceral leishmaniasis. Journal of Immunological Methods, 277 (1). 01-10.
 
Relation http://dx.doi.org/
http://www.eprints.iicb.res.in/769/