خانه / مقالات پژوهشی / علوم آزمایشگاهی (مقالات پژوهشی) / Truncated MTA-1: A Pitfall in ELISA-Based Immunoassay of HTLV-1 Infection

Truncated MTA-1: A Pitfall in ELISA-Based Immunoassay of HTLV-1 Infection

Source:

Hindawi Publishing Corporation
Journal of Biomedicine and Biotechnology

Author:

Mohammad Reza Abbaszadegan, Narges Jafarzadeh, Mojtaba Sankian, AbdolReza Varasteh, MahmoudMahmoudi, Majid Sadeghizadeh, Fatemeh Khatami, and NeemaMehramiz

Abstract

HTLV-1 causes adult T-cell leukemia (ATL) and HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP). Recombinant envelope glycoprotein is used in production of diagnostic enzyme-linked immunosorbent assay (ELISA) kit. There are some reports that a significant percentage of Iranian HTLV-1 infected patients showed no seroreactivity with MTA-1 peptide, while HTLV-1 had been confirmed by PCR detection methods or ELISA kits containing a cocktail of HTLV-1  specific peptides. This report describes experiments designed to determine whether some discrepancies between ELISA and PCR results could be due to truncation of immunodominant epitopes using immunoassaymethod.We have cloned theMTA-1 epitope of env gene from HTLV-1 in NotI/NdeI sites of pET22b(+) expression vector. Sequencing analysis of recombinant plasmids revealed an insertion of a cytosine in position 271 causing a stop codon in the MTA-1 protein translation. SDS-PAGE analysis also failed to reveal the presence of the desired protein. Subjects with a mutant HTLV-1 env gene were shown to be seronegative using ELISA, but positive with PCR.

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