Method and kit for evaluation of HIV mutations

Details Method and kit for evaluation of HIV mutations Method and kit for evaluation of HIV mutations

2001-03-27

2003-11-25

Horlick, Kenneth R. (Department: 1637)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

C435S006120, C536S024330

Reexamination Certificate

active

06653107

ABSTRACT:

BACKGROUND OF THE INVENTION
Genetic testing to determine the presence of or a susceptibility to a disease condition offers incredible opportunities for improved medical care, and the potential for such testing increases almost daily as ever increasing numbers of disease-associated genes and/or mutations are identified. A major hurdle which must be overcome to realize this potential, however, is the high cost of testing. This is particularly true in the case of highly polymorphic genes where the need to test for a large number of variations may make the test procedure appear to be so expensive that routine testing can never be achieved.
Testing for changes in DNA sequence can proceed via complete sequencing of a target nucleic acid molecule, although many persons in the art believe that such testing is too expensive to ever be routine. Changes in DNA sequence can also be detected by a technique called ‘single-stranded conformational polymorphism” (“SSCP”) described by Orita et al.,
Genomics
5: 874-879 (1989), or by a modification thereof referred to a dideoxy-fingerprinting (“ddF”) described by Sarkar et al.,
Genomics
13: 4410443 (1992). SSCP and ddF both evaluate the pattern of bands created when DNA fragments are electrophoretically separated on a non-denaturing electrophoresis gel. This pattern depends on a combination of the size of the fragments and of the three-dimensional conformation of the undenatured fragments. Thus, the pattern cannot be used for sequencing, because the theoretical spacing of the fragment bands is not equal.
This application relates to a particular test which can be useful as part of a testing protocol for the detection and characterization of human immunodeficiency virus (HIV).
SUMMARY OF THE INVENTION
The method of the invention provides a method for obtaining information about the allelic type of a sample of genetic material derived from an HIV-infected sample. A test is performed in which the sequence is determined in the 3′-direction for all four bases. This test will identify substantially all of the samples in which the sequence of the sample is determined in both the 3′ and 5-direction for all four bases.
To perform the method of the invention, reagents suitable for the tests are suitably packaged as a kit. The kit contains reagents for performing a four-base sequence determination on one or both strands of the target DNA. One-stranded sequence determination could be performed all in the 3′-direction, all in the 5′-direction, or as a combination of the two strands.


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