Unit

UNIT 12.8 In Situ Hybridization for Detection of HIV RNA

  1. Cecil H. Fox1,
  2. Michele Cottler-Fox2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im1208s06

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Fox, C. H. and Cottler-Fox, M. 2001. In Situ Hybridization for Detection of HIV RNA. Current Protocols in Immunology. 6:12.8:12.8.1–12.8.21.

Author Information

  1. 1

    Molecular Histology, Gaithersburg, Maryland

  2. 2

    National Institutes of Health, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1993

Abstract

In HIV studies, in situ hybridization can be used for identifying virion RNA, mRNA being produced for virion packaging, and proviral DNA in the cytoplasm or integrated in the nucleus. This unit focuses primarily on identifying virion RNA, because this is the most sensitive means by which in situ hybridization can be employed to detect HIV expression. In situ hybridization, as developed for HIV RNA detection, involves several protocols: (1) preparation of a radioactive or nonradioactive RNA probe; (2) in situ hybridization of probe to cells and paraffin sections of tissue; (3) detection of radiolabeled probe by emulsion autoradiography; (4) development, staining, and mounting of slides; and finally (5) examination of slides by bright-field, dark-field, specular reflectance, or laser-scanning confocal microscopy. The protocols presented in this unit describe a setup involving up to 150 slides.