In-situ hybridization to detect nucleic acid sequences in morphologically intact cells

Patent 4888278 Issued on December 19, 1989. Estimated Expiration Date:

October 13, 2008. Estimated Expiration Date is calculated based on simple USPTO term provisions. It does not account for terminal disclaimers, term adjustments, failure to pay maintenance fees, or other factors which might affect the term of a patent.

Abstract Claims Description Full Text

Patent References

Recombinant DNA process utilizing a papilloma virus DNA as a vector
Patent #: 4419446
Issued on: 12/06/1983
Inventor: Howley , et al.

Methods for the in vitro detection and identification of unknown pathogens or genetic entities Patent #: 4652517
Issued on: 03/24/1987
Inventor: Scholl , et al.

Inventors

Assignee

University of Massachusetts Medical Center

Application

No. 257198 filed on 10/13/1988

US Classes:

435/6, Involving nucleic acid435/810, PACKAGED DEVICE OR KIT436/501, BIOSPECIFIC LIGAND BINDING ASSAY436/504, Radioactive label436/527, Glass or silica436/528, Carrier is organic536/24.3Probes for detection of specific nucleotide sequences or primers for the synthesis of DNA or RNA

Examiners

Primary: Warden, Robert J.
Assistant: Wagner, Richard

Attorney, Agent or Firm

Hamilton, Brook, Smith & Reynolds

International Classes

C12N 015/00
G01N 033/566
G01N 033/552

Abstract

Improved methodologies for in-situ hybridization and detection of hybridized nucleic acid sequences in cell cultures and tissue sections are provided which offer an increase of speed, sensitivity, and simplicity unavailable in previously known techniques. The invention detects specific nucleic acids of interest, particularly RNA sequences, within cells and tissues utilizing DNA of a particular size as a probe to find those sequences which are held substantially in common between the cell or tissue and the probe. The cells are fixed preferably in paraformaldehyde and then hybridized using a hybridization fluid for not less than 10 minutes but not substantially more than 24 hours. A variety of identifying labels are attached to the probe which permit quick and rapid detection via measurement of radioactive isotope decay or by colorimetric detection of enzymatic reaction products. The invention is intended for use as a diagnostic kit in clinical/diagnostic laboratory testing facilities in that it permits a relatively unskilled person to accurately and reproducibly detect a few molecules of a specific nucleic acid of interest in-situ in 10 minutes.

Other References

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