Eastman-kodak Company

Johnson & Johnson Clinical Diagnostics, Inc.

Tobias D Ekeze

Thomas J Cummins

Brent A Burdick

This invention provides a rapid and highly effective method for extracting nucleic acids from cells or virions without the use of proteolytic enzymes. Extraction is accomplished within a few minutes using a lysing composition comprising a buffer, a source of a DNA polymerase cofactor, a stabilizer and at least one nonionic surfactant which will release nucleic acids from cytoplasmic and nuclear membranes of cells or virions. The resulting mixture is heated to boiling for up to fifteen minutes, and the nucleic acids are recovered for amplification using polymerase chain reaction. No proteolytic enzyme is used in the extraction process.

Methods of extracting deoxyribonucleic acids without using a proteolytic

A nucleic acid is rapidly extracted from whole blood or a peripheral blood mononuclear cell (PBMC) fraction thereof. Extraction from the PBMC fraction is accomplished by heating the fraction at or near the boiling point of water for a few minutes and recovering the extracted nucleic acid. This rapid method is particularly useful for extracting DNA for the detection of genetic diseases or infectious agents, such as HIV-I. Whole blood can likewise be heated after it is mixed with a salt solution containing a polysaccharide, such as dextran. The extracted nucleic acid is then recovered from the heated mixture. Nucleic acids extracted in this way are available for amplification using a polymerase chain reaction. Where the presence of a specific gene is to be determined for diagnostic purposes, it can be extracted as described above and subjected to suitable amplification and detection steps.

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Rochester, NY, United States of America

Tobias D Ekeze