Amplification and detection of nucleic acids in blood samples

Details Amplification and detection of nucleic acids in blood samples Amplification and detection of nucleic acids in blood samples

1993-09-08

1996-03-26

Jones, W. Gary

Chemistry: molecular biology and microbiology

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

Preparing compound containing saccharide radical

435 4, 435 5, 435 6, 435 911, 435267, 435269, 435270, 536 231, 536 2433, C12P 1934, C12Q 100, C12Q 148, C12Q 168

Patent

active

055019633

ABSTRACT:
This invention relates to a process for the amplification of nucleic acids in the form of DNA or RNA from blood samples by means of an enzymatic amplification method, characterized in that no preparation of the blood sample otherwise necessary to prepurify the nucleic acid to be amplified is performed and the proportion of the sample in the reaction mixture for the amplification process is greater than 5 volume % if a specific amount of salt is present in the reaction mixture. Depending on the proportion of blood sample and its salt contribution of monovalent and/or bivalent ions, the salt concentration in the reaction mixture in which the amplification is performed is, where applicable, adapted to the enzyme requirements by the use of an appropriately concentrated salt solution.

REFERENCES:
patent: 5035996 (1991-07-01), Hartley
patent: 5284940 (1994-02-01), Lin et al.
Mercier, et al., Nuc. Acids Res., 18:5908, "Direct PCR from whole blood, without DNA extraction" (1990).
Panaccio and Lew, Nuc. Acids Res., 19:1151 (1991) "PCR based diagnosis in the presence of 8% (v/v) blood".
McCusker, et al., Nuc. Acids Res., 20:6747 (1992) "Improved Method for direct PCR amplification from whole blood".
Ravaggi, et al., PCR Method and Applications, 1:291-292 (1992) "Direct PCR Amplification of HCV RNA from Human Serum".
Panaccio, et al., BioTechniques, 14:238-243 (1993) "FoLT PCR: A Simple PCR Protocol for Amplifying DNA Directly from Whole Blood".
Cimino et al. Nucl. Acids Res. 19(1):99 (1990).
Izraeli et al. Nucl. Acids Res. 19(21) (1991).

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