RESUMEN
Successful disease prevention and therapy critically depend on timely diagnosis of infections. Quantitative immuno-PCR (qiPCR) technology improves the sensitivity in the detection of antibodies to pathogens. A qiPCR-based assay was developed to determine IgG antibodies to Epstein-Barr virus (EBV) in the human blood serum. EBV nuclear protein 1 fragment (pEBV) was expressed in Escherichia coli. A synthetic single-stranded deoxyribonucleotide was conjugated to streptavidin, and the conjugate was used to detect ÑEBV-IgG1-biotin complexes by qiPCR. The IgG1 titers determined by qiPCR were compared to the results of enzyme-linked immunosorbent assay (ELISA). The sensitivity of qiPCR was one order of magnitude higher than that of ELISA. Thus, a highly sensitive qiPCR-based assay was developed to quantitate antibodies specific to the recombinant EBV antigen.
Asunto(s)
Anticuerpos/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Infecciones por Virus de Epstein-Barr/sangre , Herpesvirus Humano 4/aislamiento & purificación , Antígenos Virales/sangre , Antígenos Virales/inmunología , Infecciones por Virus de Epstein-Barr/inmunología , Herpesvirus Humano 4/inmunología , Herpesvirus Humano 4/patogenicidad , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Proteínas Nucleares/sangre , Proteínas Nucleares/inmunología , Reacción en Cadena de la PolimerasaRESUMEN
Theoretical basis and an algorithm of calculations for frequencies of genes and genotypes in the population of species with obligate cross-pollination under assessment for the character of polymorphic genetic basis are proposed. Use of four rye populations and the F1 and F2 from their crossings allows to determine genetic control of beta-amylase spectrum through identification of two genes, establishment of the allele number in each gene, and showing the linkage between beta-Amy-1 and the gene for self-incompatibility S2.