Use of rnase h for the selective amplification of viral dna
AssigneeRoche Molecular Systems, Inc.
InventorEllen H Fiss
Life of this patent
Estimating the term…
Abstract
The present invention concerns the amplification of at least a first and a second target nucleic acid that may be present in at least one fluid sample using RNase H and a polymerase with reverse transcriptase activity.
Claims12
A process for amplifying a DNA target nucleic acid that may be present in a sample, the process comprising the steps of contacting the DNA target nucleic acid from the sample with an amplification reagent comprising RNase H and a polymerase with reverse transcriptase activity;
- incubating in a reaction vessel the DNA target nucleic acid with the amplification reagent for a period of time, under conditions suitable for transcription of RNA by the polymerase with reverse transcriptase activity to occur;
- and incubating in the reaction vessel the DNA target nucleic acids with the amplification reagent for a period of time, under conditions sufficient for an amplification reaction indicative of the presence or absence of the DNA target nucleic acid to occur.
An automated process for amplifying and determining the quantity of a DNA target nucleic acid that may be present in a fluid sample, said process comprising the steps of incubating in a reaction vessel said DNA target nucleic acid with a solution comprising amplification reagents, and oligonucleotides specific for said DNA target nucleic acid, wherein said amplification reagents comprise RNase H and a polymerase with reverse transcriptase activity, for a period of time and under conditions suitable for transcription of RNA by said polymerase with reverse transcriptase activity to occur;
- incubating in the reaction vessel the DNA target nucleic acid with the solution comprising amplification reagents, and oligonucleotides specific for the DNA target nucleic acid for a period of time and under conditions sufficient for an amplification reaction indicative of the presence or absence of the DNA target nucleic acid to occur;
- and determining the quantity of the DNA target nucleic acid by referencing to an external calibration or by implementing an internal quantitative standard.
Dependencies are inferred from the claim text.