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Single-copy Gene Detection Using Branched DNA (bDNA) In Situ Hybridization
Abstract
We have developed a branched DNA in situ hybridization (bDNA ISH) method for detection of human papillomavirus (HPV) DNA in whole cells. Using human cervical cancer cell lines with known copies of HPV DNA, we show that the bDNA ISH method is highly sensitive, detecting as few as one or two copies of HPV DNA per cell. By modifying sample pretreatment, viral mRNA or DNA sequences can be detected using the same set of oligonucleotide probes. In experiments performed on mixed populations of cells, the bDNA ISH method is highly specific and can distinguish cells with HPV-16 from cells with HPV-18 DNA. Furthermore, we demonstrate that the bDNA ISH method provides precise localization, yielding positive signals retained within the subcellular compartments in which the target nucleic acid sequences are localized. As an effective and convenient means for nucleic acid detection, the bDNA ISH method is applicable to the detection of cancers and infectious agents. (J Histochem Cytochem 49:603–611, 2001)
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Cited by
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Bressan, D., Battistoni, G., & Hannon, G. J. (2023) · Science · doi:10.1126/science.abq4964 · Cited by 594
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Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes
Beliveau, B. J., Joyce, E. F., Apostolopoulos, N., Yilmaz, F., Fonseka, C. Y., McCole, R. B., Chang, Y., Li, J. B., Senaratne, T. N., Williams, B. R., et al. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1213818110 · Cited by 458
Metadata
- Authors
- Player, A. N., Shen, L.-P., Kenny, D., Antao, V. P., & Kolberg, J. A.
- Venue
- Journal of Histochemistry & Cytochemistry
- Publisher
- SAGE Publications
- Published
- 2001-05
- Volume
- 49
- Issue
- 5
- Pages
- 603-611
- ISSN
- 0022-1554 (ISSN-L)
- 1551-5044
- OpenAlex
- W2129266910
- Cited by
- 194
- References
- 0
- FWCI
- 1.30
- Type
- journal article
- Language
- en
- Open access
- closed
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- Player, A. N., Shen, L.-P., Kenny, D., Antao, V. P., & Kolberg, J. A. (2001). Single-copy Gene Detection Using Branched DNA (bDNA) In Situ Hybridization. Journal of Histochemistry & Cytochemistry. https://doi.org/10.1177/002215540104900507
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Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
