Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 14 May 1993:
Vol. 260. no. 5110, pp. 976 - 979
DOI: 10.1126/science.8493534
Prev | Table of Contents | Next

Articles

Science, Vol 260, Issue 5110, 976-979
Copyright © 1993 by American Association for the Advancement of Science

articles

Detection of HIV-1 DNA and messenger RNA in individual cells by PCR-driven in situ hybridization and flow cytometry

BK Patterson, M Till, P Otto, C Goolsby, MR Furtado, LJ McBride, and SM Wolinsky

Department of Pathology, Northwestern University Medical School, Chicago, IL 60611.

Human immunodeficiency virus type-1 (HIV-1) DNA and messenger RNA sequences in both cell lines and blood obtained directly from HIV-1-infected patients were amplified by polymerase chain reaction and hybridized to fluorescein-labeled probes in situ, and the individually labeled cells were analyzed by flow cytometry. After flow cytometric analysis, heterogeneous cell populations were reproducibly resolved into HIV-1-positive and -negative distributions. Fluorescence microscopy showed that the cellular morphology was preserved and intracellular localization of amplified product DNA was maintained. Retention of nonspecific probe was not observed. Analysis of proviral DNA and viral messenger RNA in cells in the blood of HIV-1-infected patients showed that the HIV-1 genome persists in a large reservoir of latently infected cells. With the use of this technique it is now possible to detect single-copy DNA or low-abundance messenger RNA rapidly and reproducibly in a minor subpopulation of cells in suspension at single-cell resolution and to sort those cells for further characterization.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Multitarget magnetic activated cell sorter.
J. D. Adams, U. Kim, and H. T. Soh (2008)
PNAS 105, 18165-18170
   Abstract »    Full Text »    PDF »
In-Cell PCR Method for Specific Genotyping of Genomic DNA from One Individual in a Mixture of Cells from Two Individuals: A Model Study with Specific Relevance to Prenatal Diagnosis Based on Fetal Cells in Maternal Blood.
T. V. Hviid (2002)
Clin. Chem. 48, 2115-2123
   Abstract »    Full Text »    PDF »
Direct Detection by In Situ PCR of the amoA Gene in Biofilm Resulting from a Nitrogen Removal Process.
T. Hoshino, N. Noda, S. Tsuneda, A. Hirata, and Y. Inamori (2001)
Appl. Envir. Microbiol. 67, 5261-5266
   Abstract »    Full Text »    PDF »
Direct in situ reverse transcriptase-polymerase chain reaction.
R. Kher and R. Bacallao (2001)
Am J Physiol Cell Physiol 281, C726-C732
   Abstract »    Full Text »    PDF »
Single-copy Gene Detection Using Branched DNA (bDNA) In Situ Hybridization.
A. N. Player, L.-P. Shen, D. Kenny, V. P. Antao, and J. A. Kolberg (2001)
J. Histochem. Cytochem. 49, 603-612
   Abstract »    Full Text »
Positive and Negative Aspects of the Human Immunodeficiency Virus Protease: Development of Inhibitors versus Its Role in AIDS Pathogenesis.
K. Ikuta, S. Suzuki, H. Horikoshi, T. Mukai, and R. B. Luftig (2000)
Microbiol. Mol. Biol. Rev. 64, 725-745
   Abstract »    Full Text »    PDF »
CD8+ cell noncytotoxic anti-human immunodeficiency virus response inhibits expression of viral RNA but not reverse transcription or provirus integration.
C. E. Mackewicz, B. K. Patterson, S. A. Lee, and J. A. Levy (2000)
J. Gen. Virol. 81, 1261-1264
   Abstract »    Full Text »
Applications of Flow Cytometry to Clinical Microbiology.
A. Alvarez-Barrientos, J. Arroyo, R. Canton, C. Nombela, and M. Sanchez-Perez (2000)
Clin. Microbiol. Rev. 13, 167-195
   Abstract »    Full Text »    PDF »
Cell-Mediated DNA Transport Between Distant Inflammatory Sites Following Intradermal DNA Immunization in the Presence of Adjuvant.
A. La Cava, R. Billetta, G. Gaietta, D. B. Bonnin, S. M. Baird, and S. Albani (2000)
J. Immunol. 164, 1340-1345
   Abstract »    Full Text »    PDF »
Cellular Localization of Latent Murine Cytomegalovirus.
A. J. Koffron, M. Hummel, B. K. Patterson, S. Yan, D. B. Kaufman, J. P. Fryer, F. P. Stuart, and M. I. Abecassis (1998)
J. Virol. 72, 95-103
   Abstract »    Full Text »    PDF »
In situ PCR: protocols and applications..
G J Nuovo (1995)
Genome Res. 4, S151-S167
   Abstract »    PDF »
HIV population dynamics in vivo: implications for genetic variation, pathogenesis, and therapy.
J. Coffin (1995)
Science 267, 483-489
   Abstract »    PDF »
Virologic and Immunologic Characterization of Long-Term Survivors of Human Immunodeficiency Virus Type 1 Infection.
Y. Cao, L. Qin, L. Zhang, J. Safrit, and D. D. Ho (1995)
N. Engl. J. Med. 332, 201-208
   Abstract »    Full Text »    PDF »
Multifactorial nature of human immunodeficiency virus disease: implications for therapy.
A. Fauci (1993)
Science 262, 1011-1018
   Abstract »    PDF »
Scientific and social issues of human immunodeficiency virus vaccine development.
B. Haynes (1993)
Science 260, 1279-1286
   Abstract »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)