Volume 17, Issue 3 (Autumn 2020)                   Sci J Iran Blood Transfus Organ 2020, 17(3): 200-209 | Back to browse issues page

XML Persian Abstract Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Mehrabi Habibabadi H, Sharifi Z, Parsania M, Pourfathollah A, Haghighat S. Comparison of Western blotting and PCR assays for confirmation of HTLV-1 virus in volunteer blood donors. Sci J Iran Blood Transfus Organ 2020; 17 (3) :200-209
URL: http://bloodjournal.ir/article-1-1315-en.html
Comparison of Western blotting and PCR assays for confirmation of HTLV-1 virus in volunteer blood donors
H. Mehrabi Habibabadi , Z. Sharifi * , M. Parsania , A.A. Pourfathollah , S. Haghighat
Keywords: Key words: Western Blotting, Chemiluminescence, Nested PCR, HTLV-1
Full-Text [PDF 465 kb]   (1599 Downloads)     |   Abstract (HTML)  (3030 Views)
Full-Text:   (5048 Views)
    References:
 
  1. Matsuoka M, Jeang KT. Human T-cell leukemia virus type I at age 25: a progress report. Cancer Res 2005; 65(11): 4467-70.
  2. Verdonck K, González E, Van Dooren S, Vandamme AM, Vanham G, Gotuzzo E. Human T-lymphotropic virus 1: recent knowledge about an ancient infection. Lancet Infect Dis 2007; 7(4): 266-81.
  3. Poiesz BJ, Ruscetti FW, Gazdar AF, Bunn PA, Minna JD, Gallo RC. Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma. Proc Natl Acad Sci USA 1980; 77: 7415-9.
  4. Lowis GW, Sheremata WA, Minagar A. Epidemiologic features of HTLV-II: serologic and molecular evidence. Ann Epidemiol 2002; 12(1): 46-66.
  5. Calattini S, Chevalier SA, Duprez R, Bassot S, Froment A, Mahieux R, Gessain A. Discovery of a new human T-cell lymphotropic virus (HTLV-3) in Central Africa. Retrovirology 2005; 2: 30.
  6. Mahieux R, Gessain A. The human HTLV-3 and HTLV-4 retroviruses: new members of the HTLV family. Pathol Biol (Paris) 2009; 57(2): 161-6.
  7. Vrielink H, Reesink HW. HTLV-I/II prevalence in different geographic locations. Transfus Med Rev 2004; 18(1): 46-57.
  8. Tabei SZ , Safaei A. HTLV1 infection in the world and Khorasan. J Birjand Univ Med Sci 2008; 15(1): 5-16. [Article in Farsi]
  9. Proietti FA, Carneiro-Proietti AB, Catalan-Soares BC, Murphy EL. Global epidemiology of HTLV-I infection and associated diseases. Oncogene 2005; 24(39): 6058-68.
  10. Aceijas C, Stimson GV, Hickman M, Rhodes T. Global overview of injecting drug use and HIV infection among injecting drug users. AIDS 2004; 18(17): 2295-303.
  11. Durrant I. Enhanced chemiluminescent detection of horseradish peroxidase labeled probes. Methods Mol Biol 1994; 31: 147-61.
  12. Alegria-Schaffer A, Lodge A, Vattem K. Performing and optimizing Western blots with an emphasis on chemiluminescent detection. Methods Enzymol 2009; 463: 573-99.
  13. Kapeluich YL, Rubtsova M, Egorov AM. Enhanced chemiluminescence reaction applied to the study of horseradish peroxidase stability in the course of p-iodophenol oxidation. J Biolumin Chemilumin 1997; 12(6): 299-308.
  14. Bergendahl V, Glaser BT, Burgess RR. A fast Western blot procedure improved for quantitative analysis by direct fluorescence labeling of primary antibodies. J Immunol Methods 2003;  277(1–2): 117-25.
  15. Pirayeshfard L, Sharifi Z, Amini-Kafiabad S, Haghnazari Sadaghiani N. Phylogenetic analysis of HTLV-1 in Iranian blood donors, HIV-1 positive patients and patients with beta thalassemia. J Med Virol 2018; 90(8): 1398-405.
  16. Cassar O, Charavay F, Touzain F, Jeannin P, Grangeon J-P, Laumond S, et al. A novel human T-lymphotropic virus type 1c molecular variant in an indigenous individual from New Caledonia, Melanesia. PLoS Negl
Trop Dis 2017; 11(1): e0005278.
  1. Nobre AFS, Almeida DS, Ferreira LC, Ferreira DL, Junior ECS, Viana M, et al. Low genetic diversity of the Human T-cell Lymphotropic Virus (HTLV-1) in an endemic area of the Brazilian Amazon basin. PloS One 2018; 13(3): e0194184.
  2. Xiaohui Bi, Hongxia Ning, Tingting Wang, Dongdong Li, Yongming Liu, Tingfu Yang, et al. Comparative Performance of Electrochemiluminescence Immunoassay and EIA for HIV Screening in a Multiethnic Region of China. PLoS One 2012; 7(10): e48162.
  3. Boadu R, Darko G, Nortey P, Akweongo P, Sarfo B. Assessing the sensitivity and specificity of First Response HIV-1-2 test kit with whole blood and serum samples: a cross-sectional study. AIDS Res Ther 2016; 13: 9.
  4. Weggheleire AD, An S, Baetselier I D, Soeung P, Keath H, So V, et al. A cross-sectional study of hepatitis C among people living with HIV in Cambodia: Prevalence, risk factors, and potential for targeted screening. PLoS One 2017; 12(8): e0183530
  5. Moradi A, Mansourian AR, Ahmadi Ar, Ghaemi E, Kalavi KH, Marjani A, et al. Prevalence of HTLV-I antibody among major thalassemic patients in Gorgan(south east of Caspian sea). J Appl Sci 2008; 8: 391-3. [Article in Farsi]
  6. Anaraki Mohammadi Gh.R., Sadeghipour A.R., Vossough P, Nour Mohammadi I, Mirnateghi A.M.. Assessment of the Prevalence of Human T-Lymphotropic Virus type 1 among Thalassemic Patients with Frequent Blood Transfusion in Tehran in 2003. Journal of Iran Medical science University 2005; 12(47): 19-24.
  7. Shahabi M,  Sayadpour Zanjani D, Afzal Aghaei M,  Maghsoudlou M,  Bazargani R ,  Shakibaei H,  et al. Seroepidemiology of blood donors in Khorasan Razavi province with unknown Western blot results of human T-cell lymphotropic virus types 1 and 2. Iran J Infect Dis Trop Med 2014; 19(65): 91. [Article in Farsi]
  8. Jamili P, Safabakhsh H, Sadegh marvi M, Taghi Shakeri M, Sadeghi R. Seroepidemiology of HTLV infection in Sabzevar blood donors from 2007 to 2008. Journal of Sabzevar University of Medical Sciences 2014; 4(20): 271. [Article in Farsi]
  9. Stramer SL, Townsend RL, Foster GA, Johnson R, Weixlmann B, Dodd RY. Discordant human T-lymphotropic virus screening with Western blot confirmation: evaluation of the dual-test algorithm for US blood donations. Transfusion 2018; 58(3); 638-40.
  10. Coorssen JR, Blank PS, Albertorio F, et al. Quantitative femto- to attomole immunodetection of regulated secretory vesicle proteins critical to exocytosis. Anal Biochem 2002; 307(1): 54-62.
  11. Kurien BT, Scofield RH. Western blotting. Methods 2006; 38(4): 283-93.
  12. Murphy RM, Lamb GD. Important considerations for protein analyses using antibody based techniques: down-sizing Western blotting up-sizes outcomes. J Physiol 2013; 591(23): 5823-31.
  13. Koller    A,    Watzig    H.    Precision    and    variance
components in quantitative gel electrophoresis. Electrophoresis 2005; 26(12): 2470-5.
  1. Monavari SH, Keyvani H, Mollaie H, Fazlalipour M, Sadeghi F, Salehi-Vaziri M, et al. Detection of human T-cell lymphotropic virus Type-1 among patients with malignant hematological diseases in Capital of Iran, Tehran. J Gen Mol Virol 2011; 3(5): 67-70.
  2. Saghafi M, Rezaieyazdi Z, Nabavi S, Mirfeizi Z, Sahebari M, Salari M. HTLV-1 seroprevalance in sarcoidosis. A clinical and laboratory study in northeast
    of Iran. Int J Rheum Dis 2018; 21(6): 1309-13.
  3. Birbian DJ, Sinha N, Shweta S, Akshra G. A critical review on PCR, its types and applications. International J Adv Res Biol Sci 2014; 1(7): 65-80.
  4. Quin C, Marion R, David E B, Jonathan R, Will B. Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res 1992; 20(7): 1717-23.
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Sci J Iran Blood Transfus Organ 2020;17 (3): 200-209
Original Article
 
 
 

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
Type of Study: Research | Subject: Virology
Published: 2020/10/1
Add your comments about this article : Your username or Email:
CAPTCHA
Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2025 CC BY-NC 4.0 | Scientific Journal of Iran Blood Transfus Organ

Designed & Developed by : Yektaweb