11th International AIDS Conference Vancouver, British Columbia — July 7-12, 1996

Int Conf AIDS 1996 Jul 7-12; 11:444 (abstract no. Pub.B.1041)
Pavlova IS, Lubavina I; Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia. Fax: 095-374-6751. E.mail: pavlov&ekotehno.msk.ru.

OBJECTIVE: To develop express non-instrumental immunoassay test for detection of IgG to HIV1 and HIV2 in human sera and plasma.

METHODS: Non-instrumental dot immunoassay for detection of IgG of HIV1 and HIV2 is based on the mixture of synthetic peptides of gp41 (HIV1) and gp32 (HIV2) as antigen and colloidal dye (aniline black) - protein A conjugate as a marker. Biotinylated peptides complexed with streptavidin were immobilized on the teeth of a comb made of nonporous material.The test was performed in the wells of 96-well immunological microplate. Analyzed sera were diluted 1/10, the comb teeth were inserted and incubated for 5 min. After washing with buffer an additional row of a plate teeth was inserted in colloidal dye - protein A suspension and incubated for another 5 min. The result was read visually by appearance of black spot in the area of immobilized antigen.

RESULTS: 85 positive (77 HIV1 and 8 HIV2) and 78 negative sera tested by commercial ELISA were tested by the developed test. The sensitivity was 98.6% and specificity was 100%. The duration time was about 15 min. The developed test contains all components enough for autonomous performance of 1-24 analyses. The sensitivity and specificity with colloidal dye - protein A conjugate was the same as with enzymatic conjugates with known insoluble substrates.

CONCLUSIONS: The developed test is simple to perform, economical, rapid, stable and can be used in the areas with few laboratory facilities and by personnel with little laboratory experience.

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PubB1041

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