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Calculating HIV-1 Infectious Titre Using a Virtual TCID50 Method

  • Yong Gao
  • Immaculate Nankya
  • Awet Abraha
  • Ryan M. Troyer
  • Kenneth N. Nelson
  • Andrea Rubio
  • Eric J. Arts
Part of the Methods In Molecular Biology™ book series (MIMB, volume 485)

Abstract

Studies of HIV-1 replication kinetics and fitness require an accurate determination of the level of infectious HIV-1 present in virus stocks. The standard technique for measuring the level of replication-competent infectious virus in culture supernatants or patient samples is the tissue culture dose for 50% infectivity (TCID50), which provides an accurate assessment of the level of infectious HIV-1. However, it is a time-consuming technique which typically takes two or more weeks to complete and requires PHA-stimulated PBMC from HIV-1 seronegative donors or an appropriate cell line. Thus rapid, cell-free surrogate measures for TCID50 are desirable. Here, we introduce the virtual TCID50 technique: a new cell-free method estimating a surrogate of infectious titer by comparing the reverse transcriptase activity in virus stock to that of reference viruses with a known TCID50 value. We have demonstrated that the virtual TCID50 obtained through this technique is comparable to the actual infectious TCID50. This method greatly simplifies the process of accurate HIV-1 titration and is particularly beneficial for studies which require titration of large number of HIV-1 isolates.

Keywords

HIV-1 TCID50 Virtual TCID50 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Yong Gao
    • 1
  • Immaculate Nankya
    • 1
  • Awet Abraha
    • 1
  • Ryan M. Troyer
    • 1
  • Kenneth N. Nelson
    • 1
  • Andrea Rubio
    • 1
  • Eric J. Arts
    • 1
  1. 1.Division of Infectious Diseases Department of MedicineCase Western Reserve UniversityClevelandUSA

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