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Analysis of the serum avidity

VG Vivian Glück
LT Leonid Tydykov
AM Anna-Lena Mader
AW Anne-Sophie Warda
MB Manuela Bertok
TW Tanja Weidlich
CG Christine Gottwald
JK Josef Köstler
BS Bernd Salzberger
RW Ralf Wagner
MK Michael Koller
AG André Gessner
BS Barbara Schmidt
TG Thomas Glück
DP David Peterhoff
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To determine the serum avidity [11], the previously described ELISA [10] was modified as follows (all reagents were used as described before). Sera were titrated in eight 2.5-fold serial dilutions starting at 1:40 dilution in two side by side replicates. After the serum binding step, the wells were washed ten times with 200 µl phosphate buffered saline (PBS), containing 0.1% Tween 20 (PBS-T). Thereupon, one replicate was treated with 100 µl of 1.5 M sodium thiocyanate (NaSCN) in PBS per well while the other replicate was treated with PBS. After 15 min incubation at ambient temperature, the plate was washed again with PBS-T, conjugate was added and the ELISA was continued as previously described. The calculation of the avidity index is described below (see “Data analysis and statistics”).

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