%0 Journal Article
%T Expression of TTV-ORF2 Protein for Detection of Anti-TTV IgG Antibodies in Human Sera
%A Shiwani Singh
%A Akanksha Singh
%A Dhananjay Singh Mankotia
%A Kalpana Luthra
%A Mohammad Irshad
%J Advances in Infectious Diseases
%P 223-229
%@ 2164-2656
%D 2013
%I Scientific Research Publishing
%R 10.4236/aid.2013.33033
%X <p class=\"MsoNormal\">
	<span lang=\"EN-US\">The </span><span lang=\"EN-US\">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</span><span lang=\"EN-US\">,</span><span lang=\"EN-US\"> identified as 20</span><span lang=\"EN-US\"> </span><span lang=\"EN-US\">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.</span>
</p>
%K TTV
%K ORF-2
%K Anti TTV-IgG
%K Expression
%K N-22
%U http://www.scirp.org/journal/PaperInformation.aspx?PaperID=36617