Expression of TTV-ORF2 Protein for Detection of Anti-TTV IgG Antibodies in Human Sera
Expression of TTV-ORF2 Protein for Detection of Anti-TTV IgG Antibodies in Human Sera作者机构：Clinical Biochemistry Division Department of Laboratory Medicine All India Institute of Medical Sciences New Delhi India
出 版 物：《Advances in Infectious Diseases》 (传染病进展（英文）)
年 卷 期：2013年第3卷第3期
页 面：223-229页
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
主 题：TTV ORF-2 Anti TTV-IgG Expression N-22
摘 要：The present study describes the cloning and expression of ORF-2 region of TTV genome and the use of expressed peptide in developing immunoassay for detection of anti-TTV antibodies in serum. Presence of TTV-DNA in serum was detected by PCR amplifying N-22 region of ORF-1 of TTV genome. This was followed by genotyping of TTV by RFLP using N-22 amplicon. Using genotype-1 positive serum as the source of TTV, the ORF-2 region was amplified by PCR and subsequently cloned and expressed in pET-19b vector. The expressed protein, identified as 20 kDa protein on SDS-PAGE gel, was purified by affinity chromatography and then used as antigen to develop western blot assay for detection of anti-TTV antibodies in serum. Analysis of sera for anti-TTV antibodies and their comparison with presence of TTV-DNA, produced encouraging results. There was a good relation between presence of anti-TTV and TTV-DNA in these sera samples. Anti-TTV antibodies could be detected in all TTV-DNA positive sera irrespective of the presence of TTV-genotype. This investigation demonstrates that ORF-2 peptide may be used in developing immunoassay for identification of TTV infection.
维普期刊数据库