Single tube PCR assay for detection of chromosomal mutations: application to the inversion hotspot in the factor VIII gene including optional use of subcycling PCR
US6355422B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Nov 30, 1998 |
| Grant date | Mar 12, 2002 |
| Priority date | — |
| Expiry date | Nov 30, 2018 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2600/16
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods are presented for determining the presence of an inversion in the factor VIII gene which cause hemophilia A. The methods encompass long distance, multiplex PCR (including overlapping PCR). The use of deaza-dGTP, high levels of DNA polymerases and high levels of DMSO aid in successfully performing the PCR. The use of a novel technique called subcycling PCR can also be applied as part of the methods. The technique allows for the determination of whether a person is homozygous or hemizygous for the inversion and has hemophilia A or whether a person is heterozygous for the inversion and is a carrier. The technique of long distance, multiplex PCR including use of deaza-dGTP, high levels of DNA polymerases and high levels of DMSO are applicable to the determination of the presence of other gross chromosomal aberrations such as deletions/inversions, translocations and inversions. The use of subcycling PCR can achieve efficient and more even amplification than normal two or three temperature PCR and is applicable to long distance, multiplex PCR.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.