Detection of Human T-cell Leukemia Virus Type 1 Provirus: Semiquantitative, Nested, Double Polymerase Chain Reaction

  • Chieko Matsumoto
  • Kusuya Nishioka
Part of the Frontiers of Virology book series (FRVIROLOGY, volume 1)


The polymerase chain reaction (PCR) is a powerful tool for diagnosing human T-cell leukemia virus type 1 (HTLV-I) infection, particularly for samples difficult to assess by immunological methods. We used the nested double PCR method to detect HTLV-I provirus in DNA obtained from an infected cell line and peripheral blood mononuclear cells from donated blood. This method yielded clear results even if only one template DNA was amplified. Furthermore, the amplified products showed one, two, or three bands after electro-phoresis, depending on the amounts of template DNA present. The semiquantitative, nested, double PCR and its applications are described in this chapter.


Single Polymerase Chain Reaction Amplification Stage Particle Agglutination Tropical Spastic Paraparesis Repeated Polymerase Chain Reaction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bhagavati S, Ehrlich G, Kula RW, Kwok S, Sninsky J, Udani V, Poiesz BJ (1988) Detection of human T-cell lymphoma/leukemia virus type I DNA and antigen in spinal fluid and blood of patients with chronic progressive myelopathy. N Engl J Med 318:1141–1147PubMedCrossRefGoogle Scholar
  2. Chen ISY, Slamon DJ, Rosenblatt JD, Shah NP, Quan SG, Wachsman W (1985) The x gene is essential for HTLV replication. Science 229:54–58PubMedCrossRefGoogle Scholar
  3. Duggan DB, Ehrlich GD, Davey FP, Kwok S, Sninsky J, Goldberg J, Baltrucki L, Poiesz BJ (1988) HTLV-I-induced lymphoma mimicking Hodgkin’s disease. Diagnosis by polymerase chain reaction amplification of specific HTLV-I sequences in tumor DNA. Blood 71:1027–1032PubMedGoogle Scholar
  4. Fujisawa J, Sciki M, Kiyokawa T, Yoshida M (1985) Functional activation of the long terminal repeat of human T-cell leukemia virus type I by a trans-acting factor. Proc Natl Acad Sci USA 82:2277–2281PubMedCrossRefGoogle Scholar
  5. Gessain A, Barin F, Vernant JC, Gout O, Maurs L, Calender AG (1985) Antibodies to human T-lymphotropic virus type-I in patients with tropical spastic paraparesis. Lancet 2:407–409PubMedCrossRefGoogle Scholar
  6. Green JE, Hinrichs SH, Vogel J, Jay G (1989) Exocrinopathy resembling Sjögren’s syndrome in HTLV-1 tax transgenic mice. Nature 341:72–74PubMedCrossRefGoogle Scholar
  7. Hinuma Y, Nagata K, Hanaoka M, Nakai M, Matsumoto T, Kinoshita K, Shirakawa S, Miyoshi I (1981) Adult T-cell leukemia: antigen in an ATL cell line and detection of antibodies to the antigen in human sera. Proc Natl Acad Sci USA 78:6476–6480PubMedCrossRefGoogle Scholar
  8. Ikeda M, Fujino R, Matsui T, Yoshida T, Komoda H, Imai J (1984) A new agglutination test for serum antibodies to adult T-cell leukemia virus. Gann 75:845–848PubMedGoogle Scholar
  9. Inoue J, Yoshida M, Sciki M (1987) Transcriptional (p40x) and post-transcriptional (p27x-III) regulators are required for the expression and replication of human T-cell leukemia virus type I genes. Proc Natl Acad Sci USA 84:3653–3657PubMedCrossRefGoogle Scholar
  10. Iwakura Y, Tosu M, Yoshida E, Takiguchi M, Sato K, Kitajima I, Nishioka K, Yamamoto K, Takeda T, Hatanaka M, Yamamoto H, Sekiguchi T (1991) Induction of inflammatory arthropathy resembling rheumatoid arthritis in mice transgenic for HTLV-I. Science 253:1026–1028PubMedCrossRefGoogle Scholar
  11. Kalyanaraman VS, Sarngadharan MG, Robert-Guroff M, Miyoshi I, Blayney D, Golde D, Gallo RC (1982) A new subtype of human T-cell leukemia virus (HTLV-II) associated with a T-cell variant hairy cell leukemia. Science 218:571–573PubMedCrossRefGoogle Scholar
  12. Kamihira S, Nakashima S, Oyakawa Y, Moriuti Y, Ichimaru M, Okuda H, Kanamura M, Oota T (1987) Transmission of human T cell lymphotropic virus type I by blood transfusion before and after mass screening of sera from seropositive donors. Vox Sang 52:43–44PubMedCrossRefGoogle Scholar
  13. Kira J, Koyanagi Y, Hamakado T, Itoyama Y, Yamamoto N, Goto I (1991) HTLV-II in patients with HTLV-I-associated myelopathy. Lancet 338:64–65PubMedCrossRefGoogle Scholar
  14. Kitajima I, Yamamoto K, Sato K, Nakajima Y, Nakajima T, Maruyama I, Osame M, Nishioka K (1991) Detection of HTLV-I proviral DNA and its gene expression in synovial cells in chronic inflammatory arthropathy. J Clin Invest 88:1315–1322PubMedCrossRefGoogle Scholar
  15. Komuro A, Hayami M, Fujii H, Miyahara S, Hirayama M (1983) Vertical transmission of adult T-cell leukaemia virus. Lancet i:240CrossRefGoogle Scholar
  16. Lairmore MD, Jacobson S, Gracia F, De BK, Castillo L, Larreategui M, Roberts BD, Levin PH, Blattner WA, Kaplan JE (1990) Isolation of human T-cell lymphotropic virus type 2 from Guaymi Indians in Panama. Proc Natl Acad Sci USA 87:8840–8844PubMedCrossRefGoogle Scholar
  17. Lee H, Swanson P, Shorty VS, Zack JA, Rosenblatt JD, Chen ISY (1989) High rate of HTLV-II infection in seropositive IV drug abusers in New Orleans. Science 244:471–475PubMedCrossRefGoogle Scholar
  18. Lee HH, Swanson P, Rosenblatt JD, Chen ISY, Sherwood WC, Smith DE, Tegtmeier GE, Fernando LP, Fang CT, Osame M, Kleinman SH (1991) Relative prevalence and risk factors of HTLV-I and HTLV-II infection in US blood donors. Lancet 337:1435–1439PubMedCrossRefGoogle Scholar
  19. Matsumoto C, Mitsunaga S, Oguchi T, Mitomi Y, Shimada T, Ichikawa A, Watanabe J, Nishioka K (1990) Detection of human T-cell leukemia virus type I (HTLV-I) provirus in an infected cell line and in peripheral mononuclear cells of blood donors by the nested double polymerase chain reaction method: comparison with HTLV-I antibody tests. J Virol 64:5290–5294PubMedGoogle Scholar
  20. Miyoshi I, Kubonishi I, Yoshimoto S, Akagi T, Ohtsuki Y, Shiraishi Y, Nagata K, Hinuma Y (1981) Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells. Nature 294:770–771PubMedCrossRefGoogle Scholar
  21. Nishioka K, Maruyama I, Sato K, Kitajima I, Nakajima Y, Osame M (1989) Chronic inflammatory arthropathy associated with HTLV-I. Lancet 1:441PubMedCrossRefGoogle Scholar
  22. Okochi K, Sato H, Hinuma Y (1984) A retrospective study on transmission of adult T cell leukemia virus by blood transfusion: seroconversion in recipients. Vox Sang 46:245–253PubMedCrossRefGoogle Scholar
  23. Osame M, Usuku K, Izumo S, Ijichi N, Amitani H, Igata A, Matsumoto M, Tara M (1986) HTLV-I associated myelopathy, a new clinical entity. Lancet 1:1031–1032PubMedCrossRefGoogle Scholar
  24. Poiesz BJ, Ruscetti FW, Gazdar AF, Bunn PA, Minna JD, Gallo RC (1980) 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 77:7415–7419PubMedCrossRefGoogle Scholar
  25. Reddy EP, Sandberg-Wollheim M, Mettus RV, Ray PE, DeFreitas E, Koprowski H (1989) Amplification and molecular cloning of HTLV-I sequences from DNA of multiple sclerosis patients. Science 244:529–533CrossRefGoogle Scholar
  26. Rosenblatt JD, Golde DW, Wachsman W, Giorgi JV, Jacobs A, Schmidt GM, Quan S, Gasson JC, Chen ISY (1986) A second isolate of HTLV-II associated with atypical hairy-cell leukemia. N Engl J Med 315:372–377PubMedCrossRefGoogle Scholar
  27. Sciki M, Hattori S, Hirayama Y, Yoshida M (1983) Human adult T-cell leukemia virus: complete nucleotide sequence of the provirus genome integrated in leukemia cell DNA. Proc Natl Acad Sci USA 80:3618–3622CrossRefGoogle Scholar
  28. Sugimoto M, Nakashima H, Matsumoto M, Uyama E, Ando M, Araki S (1989) Pulmonary involvement in patients with HTLV-I-associated myelopathy: increased soluble IL-2 receptors in bronchoalveolar lavage fluid. Am Rev Respir Dis 139:1329–1335PubMedGoogle Scholar
  29. Tajima K, Tominaga S, Suchi T, Kawagoe T, Komoda H, Hinuma Y, Oda T, Fujita K (1982) Epidemiological analysis of the distribution of antibody to adult T-cell leukemia-virus-associated antigen: possible horizontal transmission of adult T-cell leukemia virus. Gann 73:893–901PubMedGoogle Scholar
  30. Uchiyama T, Yodoi J, Sagawa K, Takatsuki K, Uchino H (1977) Adult T cell leukemia: clinical and hematological features of 16 cases. Blood 50:481–492PubMedGoogle Scholar
  31. Vernant JC, Buisson G, Magdeleine J, DeThore J, Jouannelle A, Vernant CN, Monplaisir N (1988) T-lymphocyte alveolitis tropical spastic paresis and Sjögren’s syndrome. Lancet 1:177PubMedCrossRefGoogle Scholar
  32. Watanabe T, Sciki M, Yoshida M (1984) HTLV type I (US isolate) and ATLV (Japanese isolate) are the same species of human retrovirus. Virology 133:238–241PubMedCrossRefGoogle Scholar
  33. Wiley CA, Nerenberg M, Cross D, Soto-Aguilar MC (1989) HTLV-I polymyositis in patient also infected with the human immunodeficiency virus. N Engl J Med 320:992–995PubMedCrossRefGoogle Scholar
  34. Yoshida T, Kobayashi S, Yamamoto N (1986) Discrepancy between gelatin particle agglutination assay and indirect immunofluorescence assay for antibodies to adult T cell leukemia associated antigen (ATLA). J Clin Exp Med (Igaku No Ayumi) 136:387–388Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1992

Authors and Affiliations

  • Chieko Matsumoto
    • 1
  • Kusuya Nishioka
    • 1
  1. 1.The Japanese Red Cross Central Blood CenterShibuya-ku, Tokyo, 150Japan

Personalised recommendations