Detection kit for guiding warfarin dosage with high precision and detection method thereof
A detection kit and detection method technology, applied in the biological field, can solve the problems of low accuracy of the kit and long time required, and achieve the effects of reducing the risk of bleeding, stabilizing the INR value, and improving the therapeutic effect.
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Embodiment 1
[0052] (1) First collect 2ml of peripheral blood from the patient, anticoagulate with EDTA, and use SnoMag TM Blood DNA Extraction Kit extracts genomic DNA, and operates in strict accordance with the kit instructions.
[0053] (2) Perform PCR amplification on the extracted genomic DNA. The PCR reaction system is 20ul: genomic DNA 1ul, 2×EasyTaq PCR Super Mix 10ul, primer (F / R 10uMol) 0.5ul, double distilled water (ddH 2 O) 8.5ul; PCR reaction conditions are: 94°C pre-denaturation for 5min, 94°C for 30s, 60°C for 30s, 72°C for 1min, a total of 35 cycles, 72°C extension for 5min, 12°C Hold for 0min. The primer sequences are as follows:
[0054] VKORC1 F:5'-TCCAGGGTTCAAGTGGTTCTC-3'
[0055] R:5'-ATTCATGCAGGGACATCTTTGG-3'
[0056] CYP4F2 F:5'-AGTCCCGGTCATCTCCCGCCAT-3'
[0057] R: 5'-CGCCAGCCTTGGAGAGACAGACA-3'
[0058] GGCX F:5'-GCAGAACAAGAAAGCAGGCCATCA-3'
[0059] R: 5'-TCTTAGACGCCAACAAAGGCTCCA-3'
[0060] CYP2C9*2F:5'-CACTGGCTGAAAGAGCTAACAGAG-3'
[0061] R: 5'-GTGATATGG...
experiment example 1
[0082] According to the method of Example 1, 105 patients were tested, and no false positive was found, indicating that the sensitivity and specificity of the method of the present invention both reached 100%.
experiment example 2
[0084] Taking the medication method of Example 1 as the experimental group, using the method of the Chinese patent application number 201310033313.2 as the control group 1, and taking the dosage of 3 mg as the control group 2, INR-related detection was performed on 124 patients. Among them, there are 42 people in the experimental group, 44 people in the control group 1, and 38 people in the control group 2.
[0085] The INR compliance rate of the experimental group on the 3rd, 5th, and 7th day was 45.2%, 61.9%, and 83.8%; the INR compliance rate of the control group 1 on the 3rd, 5th, and 7th day was 38.3%, 50.1%, and 69.4%. The INR compliance rates of group 2 on days 3, 5, and 7 were 11.0%, 22.7%, and 46.0%.
[0086] The time for the experimental group to reach the standard for the first time was 4.10±1.22 days; the time for the control group 1 to reach the standard for the first time was 6.23±2.30 days; the time for the control group 2 to reach the standard for the first tim...
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