High-speed detection of the two common paraoxonase polymorphisms Leu55-->Met and Gln192-->Arg by real-time fluorescence PCR and melting curves.
Human paraoxonase (PON1) is a calcium-dependent esterase exclusively bound to apolipoprotein A-I and clusterin, containing high-density lipoprotein (HDL) particles that hydrolyzes organophosphates and aryl esters. Several studies have indicated that PON1 can prevent low-density lipoprotein (LDL) oxidation by hydrolyzing lipid peroxides in the lipoprotein, which is the crucial first step for atherogenesis. Therefore it may protect against the development of atherosclerosis. Serum PON1 activity has been shown to be decreased in familiar hypercholesterolemia and in diseases that are associated with accelerated atherogenesis. The PON1 gene has two common coding region polymorphisms, Leu55-->Met and Gln192-->Arg. Both polymorphisms have been identified as independent risk factors for cardiovascular disease in diabetic and non-diabetic patients. We have established high-speed and easy-to-perform genotyping for the two most significant PON1 gene polymorphisms, employing the LightCycler technology and melting curves. This technique eliminates PCR contamination related to sample handling and does not require digestion of PCR products with restriction enzymes and/or fragment separation on gels.
Harangi
Mariann
M
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany.
Aslanidis
Charalampos
C
Paragh
György
G
Schmitz
Gerd
G
D019943:Amino Acid Substitution; D001120:Arginine / Q000235:genetics; D043303:Aryldialkylphosphatase; D001483:Base Sequence; D004735:Energy Transfer; D004950:Esterases / Q000097:blood / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005453:Fluorescence; D005838:Genotype; D005973:Glutamine / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D007930:Leucine / Q000235:genetics; D008715:Methionine / Q000235:genetics; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic / Q000235:genetics
D009838:Oligodeoxyribonucleotides; D005973:Glutamine; D001120:Arginine; D008715:Methionine; D004950:Esterases; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human; D007930:Leucine