Document Detail

Standardisation and comparison of serial dilution and single dilution enzyme linked immunosorbent assay (ELISA) using different antigenic preparations of the Babesia (Theileria) equi parasite.
MedLine Citation:
PMID:  12588685     Owner:  NLM     Status:  MEDLINE    
Serial dilution and single dilution enzyme linked immunosorbent assays (ELISA) were standardised and their sensitivity and specificity were compared for serodiagnosis of Babesia equi infection. The antibody titres of 24 donkey sera of known identity were determined separately by serial dilution ELISA using three different B. equi antigens namely whole merozoite (WM), cell membrane (CM) and high speed supernatant (HSS). The ratios of the optical density (OD) of known positive and known negative sera at different serum dilutions were calculated and termed as the positive/negative (P/N) ratio. The coefficients of correlation (r) were calculated between the P/N ratios at different dilutions of sera and the log10 antibody titres of the same sera were ascertained by serial dilution ELISA. The highest value of 'r' was obtained at a serum dilution of 1:200. From log10 antibody titre of sera (y) and their P/N ratio at a dilution of 1:200 (x), regression equations (y = a + bx) were calculated separately for the three antigens. Test sera were diluted to 1:200, their OD were read in duplicate wells and were converted to the P/N ratio. Antibody titres were predicted from the P/N ratio using a regression equation separately for the three antigens. Titres obtained by both ELISAs were not significantly different from each other, thus confirming that single dilution ELISA could be successfully used to replace conventional serial dilution ELISA. The sensitivity, specificity and predictive value of single dilution ELISA was validated statistically using 42 B. equi disease-positive sera and 106 B. equi disease-negative sera. The WM antigen was found to be the most sensitive with a higher predictive value for negative test sera as compared to the CM or HSS antigens. Sera positive for other equine infections including Babesia caballi showed no cross-reaction with the three B. equi antigens in ELISA, thus the test was immunologically specific. Antibody titres of 109 unknown field donkey/horse sera obtained by serial and single dilution ELISA using the WM antigen did not show any significant difference. Since the single dilution ELISA was found to be more economical, convenient, sensitive, specific than the serial dilution ELISA and has a high predictive value, it is suitable for use in sero-epidemiological studies on B. equi infections in the field.
Sanjay Kumar; Yogesh Kumar; Dharam V Malhotra; Shruti Dhar; Anil K Nichani
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Veterinary research     Volume:  34     ISSN:  0928-4249     ISO Abbreviation:  Vet. Res.     Publication Date:    2003 Jan-Feb
Date Detail:
Created Date:  2003-02-17     Completed Date:  2003-05-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9309551     Medline TA:  Vet Res     Country:  France    
Other Details:
Languages:  eng     Pagination:  71-83     Citation Subset:  IM    
Department of Veterinary Epidemiology and Preventive Medicine, College of Veterinary Sciences, CCS Haryana Agricultural University, Hisar 125 004, Haryana, India.
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MeSH Terms
Antibodies, Protozoan / analysis,  immunology
Antigens, Protozoan / immunology*
Babesia / immunology*
Babesiosis / diagnosis*,  immunology,  veterinary*
Cross Reactions
Enzyme-Linked Immunosorbent Assay / methods*
Horse Diseases / diagnosis*,  immunology,  parasitology
Logistic Models
Sensitivity and Specificity
Reg. No./Substance:
0/Antibodies, Protozoan; 0/Antigens, Protozoan

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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