Enzyme immunoassay for the detection of anti-Kaposi's sarcoma-associated herpes virus IgG antibodies based on new mosaic protein
Abstract number: 1134_03_316
Kornienko N., Gladysheva M., Ulanova T., Puzyrev V., Burkov A., Loginova L., Obriadina A.
Kaposi's sarcoma-associated herpes virus (or human herpes virus type 8 HHV-8) is a recently discovered gamma-herpesvirus associated with 4 clinical and epidemiological variants of Kaposi sarcoma (classic, endemic, iatrogenic, and acquired immunodeficiency virus-associated), primary effusion lymphoma and multicentric Castleman's disease.
The aim of this study was to learn the antigenic properties of new recombinant mosaic protein and to develop and evaluate a screening enzyme immunoassay (EIA) for the detection of anti HHV-8 IgG activity in serum specimens.
Materials and methods:
Mosaic of 2 antigenic domains from the proteins encoded by open reading frames 65 (140170 aa) and K8.1 (3262 aa) of HHV-8 was produced as GST fusion protein to develop an assay for the detection of anti HHV-8 antibodies.
Assay conditions were optimized to reduce the possibility of false positive and false negative results. To validate the specificity and sensitivity of new EIA sera from HIV-infected individuals (n = 163), children (015 years) (n = 170), patients with sexual transmitted diseases (STD) (n = 136), and from European normal blood donors (n = 1349) were tested. Serum samples from KS patients (n = 30) were initially tested as positive by immunofluorescence assay with LANA protein. All specimens were additionally tested for IgG anti-HHV-8 activity by commercially available EIA (Vecto HHV-8-IgG-strip,Russia).
27 out of 30 HHV-8-positive samples were positive on the novel EIA. Assay sensitivity was calculated at 90%. Coincidence with commercially available EIA was 98.21%. The percentages of positive reactivity in all investigated groups were as follows: 119 for health blood donors, 1.76 for children, 2.21 for STD-patients and 3.38 for HIV-infected. Specificity of the assay was around 98.2%98.8% and there were no significant differences between health donors/children and groups at highest risk of acquiring HHV8 infection.
The artificial mosaic protein used in this study demonstrated significant potential as diagnostic reagent. The new EIA is highly specific diagnostic assay for the detection of anti-HHV-8 activity in serum specimens and may be useful tool for studies of HHV-8 epidemiology.
KSHV seroprevalence in the Russian European population is low both among health donors and HIV-infected people.
|Session name:||XXIst ISTH Congress|
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