Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Reexamination Certificate
2000-01-28
2001-10-16
Stucker, Jeffrey (Department: 1648)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
C435S006120, C536S023100, C536S024300, C536S024310, C536S024320, C536S024330
Reexamination Certificate
active
06303293
ABSTRACT:
FIELD OF THE INVENTION
The present invention pertains to improved methods for detecting nucleic acid sequences in biological samples, particularly sequences derived from infectious microorganisms.
BACKGROUND OF THE INVENTION
Millions of individuals world-wide are infected with Human Immunodeficiency Virus (HIV). Consequently, HIV infection represents a serious public health concern. Spread of HIV infection via contaminated blood products means that there is a need for screening methods that can detect small amounts of HIV RNA in patient samples. Furthermore, the increasing availability of ameliorative treatments for HIV infection means that early detection of infection in a patient is vital in order to initiate appropriate therapeutic interventions.
Thus, there is a need in the art for highly sensitive detection methods for HIV that can be used in diagnosis and screening.
SUMMARY OF THE INVENTION
The present invention provides a method for reverse transcribing Human Immunodeficiency Virus (HIV) RNA in a biological sample, where the method comprises:
(a) contacting RNA derived from said sample with an oligonucleotide under conditions in which said oligonucleotide primes synthesis of DNA complementary to at least a portion of said RNA;
wherein said oligonucleotide is selected from the group consisting of
(i) 5′-CTTGTATTACTACTG-3′<SEQ ID NO 1>,
(ii) 5′-CCCTGTGGCGCC-3′<SEQ ID NO 2>,
(iii) 5′-GCGACTAGGAGAGA-3′<SEQ ID NO 3>,
(iv) 5′-CCCAGACGGTCAGT-3′<SEQ ID NO 4>, or
(v) any combination of any of the foregoing.
In another aspect, the invention provides a method for detecting the presence of Human Immunodeficiency Virus (HIV) RNA in a biological sample, where the method comprises:
(a) performing a reverse transcription reaction using as a template RNA derived from the sample and using, as a primer, an oligonucleotide complementary to a nucleotide sequence contained within the RNA to produce HIV-specific reverse transcription products,
where the primer is selected from the group consisting of:
(i) 5′-CTTGTATTACTACTG-3′<SEQ ID NO 1>,
(ii) 5′-CCCTGTGGCGCC-3′<SEQ ID NO 2>,
(iii) 5′-GCGACTAGGAGAGA-3′<SEQ ID NO 3>,
(iv) 5′-CCCAGACGGTCAGT-3′<SEQ ID NO 4>, or
(v) any combination of any of the foregoing;
(b) amplifying products of the reverse transcription reaction to produce amplification products; and
(c) detecting the amplification products;
where detection of the amplification products indicates the presence of HIV RNA in the sample.
Amplification may be carried out by any method, preferably polymerase chain reaction (PCR). The use of HIV-specific reverse transcription primers according to the invention provides a sensitive method for detecting HIV-1 and/or HIV-2 in a sample, preferably plasma.
In yet another aspect, the invention provides kits for the detection of HIV-1, HIV-2, or a combination thereof in a biological sample, where the kit comprises a reverse transcription primer selected from the group consisting of:
(a) 5′-CTTGTATTACTACTG-3′<SEQ ID NO 1>,
(b) 5′-CCCTGTGGCGCC-3′<SEQ ID NO 2>,
(c) 5′-GCGACTAGGAGAGA3′<SEQ ID NO 3>,
(d) 5′-CCCAGACGGTCAGT-3′<SEQ ID NO 4>, or
(e) any combination of any of the foregoing. The kits may additionally comprise reagents and instructions for reverse transcription, amplification, and product detection.
REFERENCES:
patent: 5478724 (1995-12-01), Morse et al.
patent: 5688637 (1997-11-01), Moncany et al.
patent: 5840480 (1998-11-01), Guertler et al.
patent: 6001558 (1999-12-01), Backus et al.
patent: 0 887 427A2 (1998-12-01), None
patent: 98/58086 (1998-12-01), None
Linnen Jeffrey M.
Patterson David R.
Puskas John A.
Song Keming
Darby & Darby
Ortho-Clinical Diagnostics, Inc.
Stucker Jeffrey
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