Virus Genes

, Volume 32, Issue 1, pp 105–113 | Cite as

Analysis of Human Immunodeficiency Virus Type 1 Integration by Using A Specific, Sensitive and Quantitative Assay Based on Real-time Polymerase Chain Reaction

  • Norio Yamamoto
  • Chika Tanaka
  • YuFeng Wu
  • Myint Oo Chang
  • Yoshio Inagaki
  • Yasunori Saito
  • Toshio Naito
  • Hitoshi Ogasawara
  • Iwao Sekigawa
  • Yasuo Hayashida
Original Article

Abstract

A novel real-time nested-PCR assay was developed to quantify integrated human immunodeficiency virus type-1 (HIV-1) DNA with high specificity and sensitivity. This assay reproducibly allowed the detection of three copies of integrated HIV DNA in a background of 100,000 cell equivalents of human chromosomal DNA. The non-specific amplification of unintegrated HIV-1 DNA was significantly inhibited in this assay and the specificity of this assay was much higher than the previously reported method. This assay showed that kinetics in viral DNA sysnthesis was cell-type dependent and that the kinetics of HIV-1 DNA integration was very rapid in Jurkat T cell line. This method may provide new insights into the integration processes and be useful in evaluating future integrase inhibitors.

Keywords

HIV-1 Integration Alu-PCR Real-time PCR Nested PCR 

Notes

Acknowledgements

We thank Dr. Naoki Yamamoto for his support and helpful discussions. This work was supported by grants from the Ministry of Education, Science and Culture and the Ministry of Health, Labor and Welfare of Japan.

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Norio Yamamoto
    • 1
  • Chika Tanaka
    • 2
  • YuFeng Wu
    • 1
  • Myint Oo Chang
    • 1
  • Yoshio Inagaki
    • 1
  • Yasunori Saito
    • 1
  • Toshio Naito
    • 2
  • Hitoshi Ogasawara
    • 3
  • Iwao Sekigawa
    • 4
  • Yasuo Hayashida
    • 2
  1. 1.Department of Molecular Virology1-5-45 YushimaBunkyo-kuJapan
  2. 2.Department of General MedicineJuntendo University School of MedicineBunkyo-kuJapan
  3. 3.Department of Internal Medicine and RheumatologyJuntendo University School of MedicineBunkyo-kuJapan
  4. 4.Institute for Environmental and Gender Specific Medicine, Juntendo University Graduate School of MedicineUrayasu-shiJapan

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