US 5,508,166 AGranted

Method for the measurement of reverse transcriptase using immobilized primer

AssigneeAsahi Kasei Kogyo Kabushiki Kaisha

InventorsMasashi Tanno, Junzo Mizoguchi, Kouichi Sano, Masuyo Nakai

Life of this patent

Estimating the term…

Loading patent…

Abstract

A method is provided for the determination of the presence of a reverse transcriptase in a sample liquid. In an aqueous medium, at least an adenine ribopolynucleotide RNA template (poly A), an immobilized oligodeoxythyminenucleotide and a biotinylated deoxyuridine triphosphate are reacted in the presence of the sample liquid. After separation of the reacted and the unreacted biotinylated deoxyuridine triphosphate from each other, the reacted or the unreacted biotinylated deoxyuridine triphosphate is detected. Also provided is a method for the determination of infection of a patient to a retrovirus. The reaction is conducted in the presence of a sample body fluid and the reverse transcriptase. After the detection of the reacted or the unreacted biotinylated deoxyuridine triphosphate, the reverse-transcriptase-neutralizing and/or inhibiting antibody titer in the sample body fluid is determined.

Claims3

Dependent claims

A method for measuring activity of a reverse transcriptase in a sample liquid,

wherein said reverse transcriptase is that produced by human immunodeficiency virus type 1 (HIV-1), human immunodeficiency virus type 2 (HIV-2), simian immunodeficiency virus (SIV), mouse mammary tumor virus, human T-cell leukemia virus type 1 (HTLV-1), Rous avian sarcoma virus, Rouse associated virus 2 (RAV-2) or hepatitis B virus (HBV),
said method comprising reacting, in an aqueous medium, at least an adenine ribonucleotide RNA template (poly A), an immobilized oligodeoxythymidine nucleotide, a biotinylated deoxyuridine triphosphate and deoxythymidine triphosphate in the presence of the sample liquid, and, after separation of the reacted and the unreacted biotinylated deoxyuridine triphosphate from each other, detecting the reacted or unreacted biotinylated deoxyuridine triphosphate by absorptiometry, wherein said deoxythymidine triphosphate is present in a molar ratio of 3-9 relative to said biotinylated deoxyuridine triphosphate.

Dependencies are inferred from the claim text.

Facts

Application: 08/207,255Filed 8 Mar 1994 · continuation
Priority: JP 2-27274911 Oct 1990
Granted: 16 Apr 1996
Term ends: Not estimatedThe source records don’t support an estimate.

Classification8 codes

G01N 33/573
Immunoassay; Biospecific binding assay; Materials therefor: for enzymes or isoenzymes
C12Q 1/48
Measuring or testing processes involving enzymes, nucleic acids or microorganisms ; Compositions therefor; Processes of preparing such…
C12Q 1/6853
Nucleic acid amplification reactions: using modified primers or templates
C12Q 1/686
Nucleic acid amplification reactions: polymerase chain reaction [PCR]