Method for detecting polymorphisms
The invention relates to a method for determining the risk of suffering a cardiovascular disease based on the presence of different polymorphisms as well as to kits for practicing the above method. The invention also relates to a method for determining the risk of suffering a cardiovascular disease by combining the absence or presence of one or more polymorphic markers in a sample from the subject with conventional risk factors for CVD as well as computer-implemented means for carrying out said method.
1. A method of detecting polymorphisms in a biological sample, comprising
(a1) assaying the biological sample isolated from a human subject for the presence of polymorphisms at position 27 within each of the nucleic acid sequences of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10 by direct sequencing of nucleic acids in the sample;
or
(a2) assaying the biological sample isolated from a human subject for the presence of polymorphisms at position 27 within each of the nucleic acid sequences of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10 by detecting hybridization or lack of hybridization of the nucleic acid sequences of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10 in the biological sample with the set of nucleic acid probes; and
(b) detecting polymorphisms in the biological sample at position 27 within the nucleic acid sequences of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10 based on the sequencing in (a1) or detecting hybridization or lack of hybridization in (a2);
wherein the polymorphism at position 27 within the nucleic acid sequences of SEQ ID NOs:1, 3 and 7-10 is a C in SEQ ID NO:1, C in SEQ ID NO:3, T in SEQ ID NO:7, C in SEQ ID NO:8, C in SEQ ID NO:9, and T in SEQ ID NO:10.
2. The method according to claim 1 , further comprising determining one or more of age, race, sex, body mass index, blood pressure, smoking status, low density lipoprotein (LDL)- or high density lipoprotein (HDL)-cholesterol level, systolic blood pressure, diastolic blood pressure, history of heart failure, diabetes, renal insufficiency, left ventricular hypertrophy, alcohol consumption history, smoking history, exercise history, diet, and family history of cardiovascular disease or disorder of the human subject.
3. The method according to claim 1 , wherein the sample is an oral tissue sample, scraping, or wash, a biological fluid sample, saliva, urine or blood.
4. The method according to claim 1 , further comprising amplifying nucleic acids in the sample, wherein the presence or absence of the polymorphisms is identified by amplifying or failing to amplify an amplification product from the sample, wherein the amplification product is preferably digested with a restriction enzyme before analysis and/or wherein the polymorphisms are identified by hybridizing the nucleic acid sample with a primer label which is a detectable moiety.
5. The method according to claim 1 , wherein the method comprises assaying the biological sample for the presence of polymorphisms at position 27 within each of the nucleic acid sequences of SEQ ID NO:1 to 11.
6. The method according to claim 5 , wherein said polymorphisms at said positions 27 are selected from the group of C in SEQ ID NO:1, A in SEQ ID NO:2, C in SEQ ID NO:3, T in SEQ ID NO:4, C in SEQ ID NO:5, A in SEQ ID NO:6, T in SEQ ID NO:7, C in SEQ ID NO:8, C in SEQ ID NO:9, T in SEQ ID NO:10 and T in SEQ ID NO:11.