Warfarin individual anticoagulant pharmacogenomics detection kit suitable for Chinese population

Abstract

The invention provides a warfarin individual anticoagulant pharmacogenomics detection kit suitable for Chinese population, which mainly comprises CYP2C9, VKORC1 and CYP4F2 related gene-type amplification primers, CYP2C9, VKORC1 and CYP4F2 related gene-type sequencing primers, a polymerase chain reaction (PCR) reagent and a sequencing reagent. The warfarin individual anticoagulant pharmacogenomicsdetection kit suitable for the Chinese population also can comprise a specification used for explaining a warfarin pharmacogenomics dose prediction model suitable for the Chinese population or / and a computer readable storage medium recording the warfarin pharmacogenomics dose prediction model suitable for the Chinese population. The warfarin pharmacogenomics detection kit provided by the invention is simple in preparation and is convenient in use. By adopting the kit, the warfarin anticoagulant therapy dose of Chinese patient population can be accurately estimated through detecting warfarin pharmacogenomics indexes, integrating clinical environmental factors and utilizing warfarin pharmacogenomics dose prediction model software suitable for the Chinese population, which is established by the invention.

Description

technical field

[0001] The invention relates to a kit, in particular to a pharmacogenomics detection kit suitable for individualized anticoagulation of warfarin in Chinese population. Background technique

[0002] Warfarin is the most commonly used and most widely used effective oral anticoagulant drug in clinical practice. However, due to the narrow therapeutic window, the therapeutic dose varies significantly among individuals. There is a possibility of serious bleeding side effects, and insufficient dose will lead to the risk of thromboembolism due to insufficient anticoagulation. Therefore, the individualization of warfarin anticoagulant therapy has always been the focus and difficulty in clinical work.

[0003] In recent years, more and more evidences have shown that in addition to clinical environmental factors (such as age, body weight, intake of vitamin K in the diet, anticoagulant indications, concomitant medications, etc.), the genetic variation of the genome large...

Examples

Embodiment 1

[0029] Example 1: Collection of Warfarin Treatment Cases in the Chinese Population and Collection of Whole Blood DNA

[0030] 1. Collection and grouping of warfarin stable treatment cases: collect Chinese patients who have been treated with warfarin stable anticoagulant therapy for at least three months, exclude patients younger than 18 years old, and those diagnosed with cancer, liver and kidney insufficiency, or Patients with congestive heart failure.

[0031] 2. Collect clinical data related to warfarin treatment and dose, including basic clinical information of the patient (age, sex, height, weight, etc.), medical history related to warfarin dose and side effects, indications for warfarin treatment, The target prothrombin international normalized ratio (PT-INR) value, and the concomitant medication that interacts with warfarin, etc. When the INR reached a stable value for at least 3 months, the average daily dose of warfarin during stable therapy was recorded. Please ref...

Embodiment 2

[0036] Example 2: Detection of genetic variants using sanger sequencing

[0037] Design synthetic primers at both ends of the nucleotide sequence fragment at the position of the gene variant to be detected, amplify the fragment by polymerase chain reaction (PCR), and then apply Sanger sequencing to sequence and analyze the fragment, and pass the reference Sequence comparison to find the genetic variant of the site to be tested.

[0038] 1. Design synthetic primers to amplify the site of the target gene fragment and the surrounding known DNA sequence of the mutant CYP2C9*3, VKORC1 (-1639G>A) and CYP4F2 (rs2108622).

[0039] Amplification primers:

[0040] CYP2C9*3 (303bp)

[0041] Upstream primer: 5'-CCCCTGAATTGCTACAACAAA-3' (SEQ ID NO: 1)

[0042] Downstream primer: 5'-GGGACTTCGAAAACATGGAG-3' (SEQ ID NO: 2)

[0043] VKORC1(-1639G>A)(290bp)

[0044] Upstream primer: 5'-GCCAGCAGGAGAGGGAAATA-3' (SEQ ID NO: 3)

[0045] Downstream primer: 5'-AGTTTGGACTACAGGTGCCT-3' (SEQ ID NO...

Embodiment 3

[0068] Embodiment 3: the preparation and use method of kit

[0069] 1. Components of the kit

[0070] The main components of this kit are amplification primers and sequencing primers described in Example 2, PCR reaction reagents: HotStarTaq Master Mix Kit, sequencing reagents: Terminator v3.1Cycle Sequencing Kit, which can also include instructions for describing the pharmacogenomics dose prediction model of warfarin suitable for the Chinese population or / and computer-readable storage with the pharmacogenomics dose prediction model of warfarin recorded for the Chinese population medium.

[0071] 2. Storage conditions of the kit

[0072] The dry powder primers in the kit can be stored at room temperature for 2 months, and can be stored at -20°C for a long time. Dry powder primers can be stored stably at -20°C for 6 months after dissolving, and can be stored at room temperature for 120 hours when used frequently in a short period of time. HotStarTaq Master Mix Kit-20℃ can b...

technical field

[0001] The invention relates to a kit, in particular to a pharmacogenomics detection kit suitable for individualized anticoagulation of warfarin in Chinese population. Background technique

[0002] Warfarin is the most commonly used and most widely used effective oral anticoagulant drug in clinical practice. However, due to the narrow therapeutic window, the therapeutic dose varies significantly among individuals. There is a possibility of serious bleeding side effects, and insufficient dose will lead to the risk of thromboembolism due to insufficient anticoagulation. Therefore, the individualization of warfarin anticoagulant therapy has always been the focus and difficulty in clinical work.

[0003] In recent years, more and more evidences have shown that in addition to clinical environmental factors (such as age, body weight, intake of vitamin K in the diet, anticoagulant indications, concomitant medications, etc.), the genetic variation of the genome large...

Examples

Embodiment 1

[0029] Example 1: Collection of Warfarin Treatment Cases in the Chinese Population and Collection of Whole Blood DNA

[0030] 1. Collection and grouping of warfarin stable treatment cases: collect Chinese patients who have been treated with warfarin stable anticoagulant therapy for at least three months, exclude patients younger than 18 years old, and those diagnosed with cancer, liver and kidney insufficiency, or Patients with congestive heart failure.

[0031] 2. Collect clinical data related to warfarin treatment and dose, including basic clinical information of the patient (age, sex, height, weight, etc.), medical history related to warfarin dose and side effects, indications for warfarin treatment, The target prothrombin international normalized ratio (PT-INR) value, and the concomitant medication that interacts with warfarin, etc. When the INR reached a stable value for at least 3 months, the average daily dose of warfarin during stable therapy was recorded. Please ref...

Embodiment 2

[0036] Example 2: Detection of genetic variants using sanger sequencing

[0037] Design synthetic primers at both ends of the nucleotide sequence fragment at the position of the gene variant to be detected, amplify the fragment by polymerase chain reaction (PCR), and then apply Sanger sequencing to sequence and analyze the fragment, and pass the reference Sequence comparison to find the genetic variant of the site to be tested.

[0038] 1. Design synthetic primers to amplify the site of the target gene fragment and the surrounding known DNA sequence of the mutant CYP2C9*3, VKORC1 (-1639G>A) and CYP4F2 (rs2108622).

[0039] Amplification primers:

[0040] CYP2C9*3 (303bp)

[0041] Upstream primer: 5'-CCCCTGAATTGCTACAACAAA-3' (SEQ ID NO: 1)

[0042] Downstream primer: 5'-GGGACTTCGAAAACATGGAG-3' (SEQ ID NO: 2)

[0043] VKORC1(-1639G>A)(290bp)

[0044] Upstream primer: 5'-GCCAGCAGGAGAGGGAAATA-3' (SEQ ID NO: 3)

[0045] Downstream primer: 5'-AGTTTGGACTACAGGTGCCT-3' (SEQ ID NO...

Embodiment 3

[0068] Embodiment 3: the preparation and use method of kit

[0069] 1. Components of the kit

[0070] The main components of this kit are amplification primers and sequencing primers described in Example 2, PCR reaction reagents: HotStarTaq Master Mix Kit, sequencing reagents: Terminator v3.1Cycle Sequencing Kit, which can also include instructions for describing the pharmacogenomics dose prediction model of warfarin suitable for the Chinese population or / and computer-readable storage with the pharmacogenomics dose prediction model of warfarin recorded for the Chinese population medium.

[0071] 2. Storage conditions of the kit

[0072] The dry powder primers in the kit can be stored at room temperature for 2 months, and can be stored at -20°C for a long time. Dry powder primers can be stored stably at -20°C for 6 months after dissolving, and can be stored at room temperature for 120 hours when used frequently in a short period of time. HotStarTaq Master Mix Kit-20℃ can b...

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