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Sci J Iran Blood Transfus Organ 2020;17 (3): 200-209
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Comparison of Western blotting and PCR assays for confirmation of HTLV-1 virus in volunteer blood donors
Mehrabi Habibabadi H.1 , Sharifi Z.2, Parsania M.3, Pourfathollah A.A.4, Haghighat S.1
1Department of Microbiology, Faculty of Advanced Science and Technology, Tehran Medical Sciences , Islamic Azad University ,Tehran, Iran
2Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
3Department of Microbiology, Faculty of Medicine, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
4Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Background and Objectives
Human T-cell lymphotropic virus type I (HTLV-I) has been associated with two diseases in humans such as adult T-cell leukemia/lymphoma (ATLL) and HTLV-associated myelopathy/tropical spastic Para paresis (HAM/TSP). About 5 to 10 million people are infected with HTLV-1 worldwide. In order to improve the blood safety and to prevent false positive results in blood donors, methods for identifying infectious agents need to be highly sensitive and specific. The purpose of this study was to compare the electrochemical luminescence, western blotting, and PCR assays to confirm HTLV-1 virus.
Materials and Methods
This exprimental study was carried out in 2018 on 66 samples (62 males and 4 females) which had antibody against HTLV-1 virus. All retested samples with the same ELISA kit were examined by electrochemical luminescence and western blotting. For virus detection by PCR method, blood donors were recalled for new samples. After DNA extraction, nested PCR was performed with primers in both TAX and LTR regions.
Results
In this study 8 (%12/1) samples from 66 samples were reacted with ELISA test. Also 4 (%6) samples had antibody against HTLV-1 by electrochemiluminescence test. Finally, four samples were confirmed by Western blotting and Nested PCR.
Conclusions
The use of the electrochemical luminescence, western blotting and PCR assays, which had similar results in this study, is suitable for detection and confirmation of HTLV-1 virus.
Key words: Western Blotting, Chemiluminescence, Nested PCR, HTLV-1
Received: 22 Oct 2019
Accepted: 15 Feb 2020
Correspondence: Sharifi Z., PhD of Virology. Professor of Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine.
P.O.Box: 14665-1157, Tehran, Iran. Tel: (+9821) 82052152; Fax: (+9821) 88601555
E-mail:
sharifiz@yahoo.com