Measurement of Antibodies to Japanese Encephalitis Virus by the Enzyme-Linked Immunosorbent Assay Using a Small Quantity of Blood Absorbed on Filter Paper Discs

YAMAMOTO Tadao, TAKAGI Mitsuo

doi:10.11150/kansenshogakuzasshi1970.59.1135

Antibodies to Japanese encephalitis virus were assayed by the ELISA system using filter paper discs that contained human whole blood. A small quantity of blood was absorbed on a sheet of the blood sampling paper and brought to dryness. Paper discs were prepared by making holes in a blood infiltrated area of paper with a punch. Antibodies were eluted from paper discs which were placed in wells of ELISA plate containing the diluent solution, followed by the ordinary assay procedure after a removal of paper discs. The variation of measurement was minimized by vibrating the plate for 1 minute or at the time of the addition of paper discs and in the middle of the incubation period of 1 hr. Antibody titers were not reduced, when blood-absorbed filter paper was packed in a plastic bag and stored in a freezer. Antibody titers were reduced by 5 and 16% after 10 and 60 days, respectively, of storage in a refrigerator. When the specimens were stored at room temperature, the titers were decreased by 21 and 37% after 10 and 60 days, respectively, of storage.<BR>Reproducibility of the ELISA system performed with paper discs was tested, showing the correlation coefficient of 0.98 between two trials. Furthermore, correlation coefficients were 0.91 to 0.94, when antibody titers were compared between this ELISA system, the ordinary ELISA system, and the hemagglutination inhibition test. Correlation coefficients between the neutralization and other tests were 0.81 to 0.87.

Measurement of Antibodies to Japanese Encephalitis Virus by the Enzyme-Linked Immunosorbent Assay Using a Small Quantity of Blood Absorbed on Filter Paper Discs

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