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CCD Camera Detection of HIV Infection

  • John R. Day
Part of the Methods in Molecular Biology™ book series (MIMB, volume 503)

Rapid and precise quantification of the infectivity of HIV is important for molecular virologic studies, as well as for measuring the activities of antiviral drugs and neutralizing antibodies. An indicator cell line, a CCD camera, and image-analysis software were used to quantify HIV infectivity. The cells of the P4R5 line, which express the receptors for HIV infection as well as β-galactosidase under the control of the HIV-1 long terminal repeat, were infected with HIV and then stained 2 days later with X-gal to turn the infected cells blue. Digital images of monolayers of the infected cells were captured using a high resolution CCD video camera and a macro video zoom lens. A software program was developed to process the images and to count the blue-stained foci of infection. The described method allows for the rapid quantification of the infected cells over a wide range of viral inocula with reproducibility, accuracy and at relatively low cost.

Keywords

Charge-coupled device CCD Imaging HIV Infectivity Quantify Rapid 

Notes

Acknowledgments

We thank the coauthors of the original publication, Laura Martìnez, Roman Šášik, Douglas Hitchin, Megan Dueck, Douglas Richman and John Guatelli for their contributions to this work. The system we developed was conceived with ideas from Chris Aiken. We are also grateful to Prentice Higley, Sherry Rostami and Nanette Van Damme for assistance during setup and evaluation. This work was supported by grants AI27670, AI043638, AI038201, the UCSD Center for AIDS Research (AI 36214), AI29164, AI047745, from the National Institutes of Health and the Research Center for AIDS and HIV Infection of the San Diego Veterans Affairs Healthcare System.

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • John R. Day
    • 1
  1. 1.Gen-Probe IncorporatedSan DiegoUSA

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