Comparison of in-house biotin-avidin tetanus IgG enzyme-linked-immunosorbent assay (ELISA) with gold standard in vivo mouse neutralization test for the detection of low level antibodies.
Detection of anti-tetanus antibody levels is necessary for both determination of the immune status of individuals and also for planning preventive measures. ELISA is the preferred test among in vitro tests however it can be affected by the cross reacting antibodies. A previously developed in-house ELISA test was found not reliable for the antibody levels ≤1.0IU/ml. A new method was developed to detect low antibody levels correctly. The aim of the present study was to compare the results of the newly developed in-house biotin-avidin tetanus IgG ELISA test with the in vivo mouse neutralization test, for the antibody levels ≤1.0IU/ml.



A total of 54 serum samples with the antibody levels of three different levels, =0.01IU/ml, 0.01-0.1IU/ml, 0.1-1IU/ml, which were detected by in vivo mouse neutralization test were studied by the newly developed in-house biotin-avidin tetanus IgG ELISA test. Test was validated by using five different concentrations (0.01IU/ml, 0.06IU/ml, 0.2IU/ml, 0.5IU/ml, 1.0IU/ml).



A statistically significant correlation (r2=0.9967 p=0,001) between in vivo mouse neutralization test and in-house biotin-avidin tetanus IgG ELISA test, was observed. For the tested concentrations intra-assay, inter-assay, accuracy, sensitivity, specificity and coefficients of variations were determined as ≤15%.



In-house biotin-avidin tetanus IgG ELISA test can be an alternative method to in vivo mouse neutralization method for the detection of levels ≤1.0IU/ml. By using in-house biotin-avidin tetanus IgG ELISA test, individuals with non protective levels, will be reliably detected.
Public Health Institution of Turkey, Microbiology Reference Laboratories, Ankara, Turkey. Electronic address: cemilesonmez2004@yahoo.com.
Cemile
C
Sonmez
Diskapi Yildirim Beyazit Training and Research Hospital, Medical Microbiology Department, Ankara, Turkey.
Nilay
N
Coplu
Ataturk Training and Research Hospital, Medical Microbiology Department, Ankara, Turkey.
Aysegul
A
Gozalan
Public Health Institution of Turkey, Microbiology Reference Laboratories, Ankara, Turkey.
Lutfu
L
Akin
Ankara Training and Research Hospital, Medical Microbiology Department, Ankara, Turkey.
Berrin
B
Esen
D000818:Animals; D000906:Antibodies / Q000097:blood* / Q000276:immunology; D001360:Avidin / Q000276:immunology*; D001710:Biotin / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards*; D005260:Female; D007074:Immunoglobulin G / Q000276:immunology*; D008297:Male; D051379:Mice
D000906:Antibodies; D007074:Immunoglobulin G; D001360:Avidin; D001710:Biotin
Anti-tetanus antibody; Biotin avidin ELISA; In vivo mouse neutralization test; Tetanus