Abstract
The branched DNA (bDNA) technology has been used quite extensively in the microwell format to determine the levels of viral nucleic acids, as well as specific cellular RNAs [1, 2]. The bDNA technology, which is based on signal amplification, is easy to use, and does not require the use of radioactivity. With the microwell format, it has been shown to be sensitive and specific. The technology offers the additional benefits of a wide dynamic range, as well as accurate and precise quantification. We have recently applied the bDNA technology to the in situ detection of nucleic acids. In situ hybridization allows for the identification of individual cells that have a higher (or lower) level of a particular nucleic acid sequence, in the background of many cells that have normal levels. In addition, in situ hybridization also gives the morphological context of these “positive” cells, which in many applications is very important.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Kolberg JA, Ludtke DN, Shen L-P, Cao W, O’Conner D, Urdea MS, Wuestehube LJ, Lewis M. Branched DNA (bDNA) technology for direct quantification of nucleic acids: design and performance. In: Ferre F, ed. Gene Quantification. Boston: Birkhauser, 1998.
Shen L-P, Sheridan P, Cao W, Dailey PJ, Salazar-Gonzalez JF, Breen EC, Fahey JL, Urdea MS, Kolberg JA. Quantification of cytokine mRNA in peripheral blood mononuclear cells using branched DNA (bDNA) technology. J Immunol Methods 1998; 215: 123–134.
Collins M, Irvine B, Tyner D, Fine E, Zayati C, Chang C, Horn T, Ahle D, Detmer J, Shen L-P, Kolberg J, Bushnell S, Urdea M, Ho D. A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/ml. Nucl Acids Res 1997; 25: 2979–2984.
Barr S, Fusaro R, Gorrin G, Hendricks D, Kolberg J, Robertson J, Shen L, Irvine B. Presented at the 12th World AIDS Conference, Geneva, Switzerland, June 28–July 3, 1998. Clin Sci 1998; 2: 647–651.
Cao W, Collins M, Ng S, Chang C, Thompson P, Kolberg J. Poster presented at the 12th Annual San Diego Conference, The American Association for Clinical Chemistry, November 6–8, 1997, San Diego, CA.
Haase AT, Henry K, Zupanic M, Sedgewick G, Faust RA, Melroe H, Cavert W, Gebbard K, Staskus K, Zhang Z-Q, Dailey PJ, Balfour HHJ, Erice A, Perelson AS. Quantitative image analysis of HIV-1 infection in lymphoid tissue. Science 1996; 274: 985–989.
Harper ME, Marselle LM, Gallo RC, Wong-Staal F. Detection of lymphocytes expressing human T-lymphotropic virus type III in lymph nodes and peripheral blood from infected indivivals by in situ hybridization. Proc Natl Acad Sci USA 1986; 83: 772–776.
Cao W, Connolly J, Zagala M, Beard C, Hirsch A, Ku L, Kolberg J, Teramoto Y. Proceedings of the American Association for Cancer Research Eighty-Ninth Annual Meeting, New Orleans, LA. 1998; 39: 335.
Zhou J, Mulshine JL, Unsworth EJ, Scott FM, Avis IM, Vos MD, Treston AM. Purification and characterization of a protein that permits early detection of lung cancer. Identification of heterogeneous nuclear ribonucleoprotein A2/B1 as the antigen for monclonal antibody 703D4. J Biol Chem 1996; 271: 10760–10766.
Tockman MS, Mulshine JL, Piantadosi S, Erozan YS, Gupta PK, Ruckdeschel JC, Taylor PR, Zhukov T, Zhou W-H, Qiao Y-L, Yao S-X for the Lung Cancer Early Detection Work Group Investigators and the Yunnan Tin Corporation Investigators. Prospective detection of preclinical lung cancer: results from two studies of heterogeneous nuclear ribonucleoprotein A2B 1 overexpression. Clin Cancer Res 1997; 3: 2237–2246.
Grogan T, et al. 1999;(manuscript in preparation).
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Springer Science+Business Media New York
About this chapter
Cite this chapter
Antao, V.P., Player, A.N., Kolberg, J.A. (2000). In Situ Hybridization Using the bDNA Technology. In: Patterson, B.K. (eds) Techniques in Quantification and Localization of Gene Expression. Birkhäuser, Boston, MA. https://doi.org/10.1007/978-1-4612-1342-0_6
Download citation
DOI: https://doi.org/10.1007/978-1-4612-1342-0_6
Publisher Name: Birkhäuser, Boston, MA
Print ISBN: 978-1-4612-7103-1
Online ISBN: 978-1-4612-1342-0
eBook Packages: Springer Book Archive