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[Estimation of IgG Adenovirus antibody titers using a standard curve and an indirect ultramicroELISA essay].

J Laferté, Clara Savón, Ángel Goyenechea, Vanessa Vázquez, Anselmo Otero, Y Tejero, Barbara Hernández

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Abstract

It was possible to standardize a procedure which combined an indirect microELISA assay with a standard curve and that allowed to estimate the titre of IgG antibodies to Adenovirus in samples of human serum, using only one dilution of these. Based on the end-point titre previously determined for a panel of 117 serum samples, we selected 90 of these samples (r2 = 0.98) to build 4 standard curves that related the natural logarithm of the fluorescence responses to the natural logarithm of the end-point titre for a wide range of serum dilutions (1:40 = 1:320). It was selected the curve corresponding to serum dilution 1:40 (r2 = 0.81), which made possible an optimum utilization of those accessories designed to handle the volumes in the ultramicro range and, therefore, the automation of the whole procedure. The results obtained as regards the complement fixation test (100% of sensitivity and 97.3% of specificity) support the use of this method in our laboratory as a complementary tool to carry out seroepidemiological studies on a large scale and with diagnostic ends.

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It was possible to standardize a procedure which combined an indirect microELISA assay with a standard curve and that allowed to estimate the titre of IgG antibodies to Adenovirus in samples of human serum, using only one dilution of these. Based on the end-point titre previously determined for a panel of 117 serum samples, we selected 90 of these samples (r2 = 0.98) to build 4 standard curves that related the natural logarithm of the fluorescence responses to the natural logarithm of the end-point titre for a wide range of serum dilutions (1:40 = 1:320). It was selected the curve corresponding to serum dilution 1:40 (r2 = 0.81), which made possible an optimum utilization of those accessories designed to handle the volumes in the ultramicro range and, therefore, the automation of the whole procedure. The results obtained as regards the complement fixation test (100% of sensitivity and 97.3% of specificity) support the use of this method in our laboratory as a complementary tool to carry out seroepidemiological studies on a large scale and with diagnostic ends.

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Available abstract

It was possible to standardize a procedure which combined an indirect microELISA assay with a standard curve and that allowed to estimate the titre of IgG antibodies to Adenovirus in samples of human serum, using only one dilution of these. Based on the end-point titre previously determined for a panel of 117 serum samples, we selected 90 of these samples (r2 = 0.98) to build 4 standard curves that related the natural logarithm of the fluorescence responses to the natural logarithm of the end-point titre for a wide range of serum dilutions (1:40 = 1:320). It was selected the curve corresponding to serum dilution 1:40 (r2 = 0.81), which made possible an optimum utilization of those accessories designed to handle the volumes in the ultramicro range and, therefore, the automation of the whole procedure. The results obtained as regards the complement fixation test (100% of sensitivity and 97.3% of specificity) support the use of this method in our laboratory as a complementary tool to carry out seroepidemiological studies on a large scale and with diagnostic ends.

Key concepts: Serial dilution, Titer, Standard curve, Complement fixation test, Logarithm, Mathematics, Antibody, Dilution

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