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Primer set, kit and evaluation method for evaluating tacrolimus metabolism

A tacrolimus and primer set technology, applied in the field of molecular biology, can solve problems such as high serum drug concentration, time delay for tacrolimus to reach a stable therapeutic concentration, and acute poisoning reactions

Active Publication Date: 2021-02-05
JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large individual differences in the expression of tacrolimus-related metabolic enzymes in the human body, even if the same initial tacrolimus dosage corrected by kilogram body weight is given, there are still some patients after renal transplantation. It may only maintain a low serum drug concentration, resulting in a delay in the time for tacrolimus to reach a stable therapeutic concentration, thereby exposing the patient to a greater risk of acute rejection, or the serum drug concentration is too high, causing acute renal transplantation. poisoning reaction

Method used

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  • Primer set, kit and evaluation method for evaluating tacrolimus metabolism
  • Primer set, kit and evaluation method for evaluating tacrolimus metabolism
  • Primer set, kit and evaluation method for evaluating tacrolimus metabolism

Examples

Experimental program
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Effect test

Embodiment 1

[0095] The study included 220 recipients who underwent kidney transplantation in our center (the Kidney Transplant Center of the First Affiliated Hospital of Nanjing Medical University). The study design was approved by the Ethics Committee of the First Affiliated Hospital of Nanjing Medical University.

[0096] Using the present invention, predicting the blood drug concentration of tacrolimus / dose ratio per unit body weight of a patient comprises the following steps:

[0097] Step 1. Determine the basic information of the patient

[0098] The patient, a 35-year-old male, did not experience delayed recovery of renal function after surgery.

[0099] Step 2. Determine the genotype of the patient's 5 loci

[0100] (1) Extraction of total RNA from blood samples:

[0101] Take 2ml whole blood sample, add 1mL Trizol, mix quickly in ice bath, transfer to 1.5mL centrifuge tube, let stand on ice for 5min;

[0102] Add 0.2mL chloroform, shake vigorously by hand for 15 seconds to mix,...

Embodiment 2

[0143] Follow-up comparison of tacrolimus plasma concentration in 60 kidney transplant recipients 30 days after operation (see Table 5 for general information of patients), including:

[0144] (1) Extract DNA samples from peripheral blood of 60 patients, and use qRT-PCR method to detect five loci in the prediction formula (rs15524, rs4646453, rs1135612, rs2286822, rs1057868).

[0145] (2) Assignment, assign according to the following conditions.

[0146] female, gender=0, male, gender=1;

[0147] Delayed recovery of renal function did not occur, delayed recovery of renal function = 0; occurred, delayed recovery of renal function = 1;

[0148] When rs15524 genotype is TT, rs15524 genotype=0; when TC, rs15524 genotype=1; when CC, rs15524 genotype=2;

[0149] When rs4646453 genotype is TT, rs4646453 genotype=0; when TC, rs4646453 genotype=1; when CC, rs4646453 genotype=2;

[0150] When rs1135612 genotype is CC, rs1135612 genotype=0; when CT, rs1135612 genotype=1; when TT, rs11...

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Abstract

The invention belongs to the field of serum drug concentration monitoring after renal transplantation, and discloses a primer set, a kit and an evaluation method for evaluating tacrolimus metabolism.Specifically, genotypes of five SNP loci (CYP3A5 rs15524, CYP3A5 rs4646453, POR rs2286822, POR rs1135612 and POR rs1057868) on two important genes CYP3A5 and POR related to tacrolimus metabolism are detected to predict individual metabolic conditions of tacrolimus in a patient, so as to provide medication guidance for individualized design of an initial dosage of tacrolimus.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to the monitoring of serum drug metabolism state after kidney transplantation. Specifically, it relates to a primer set, a kit and an evaluation method for evaluating tacrolimus drug metabolism. Background technique [0002] Tacrolimus is one of the most commonly used immunosuppressants after renal transplantation, but its effective concentration range is narrow, and there are huge differences in pharmacokinetics among different patients. A series of complications including chronic rejection and transplant nephrotoxicity, therefore, the clinical application of tacrolimus has been greatly restricted. [0003] At present, monitoring of serum drug concentration is widely used in individualized concentration monitoring in the use of tacrolimus to minimize the side effects of tacrolimus while maintaining a sufficient therapeutic dose of tacrolimus serum drug concentration. reaction. [0...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6883C12Q1/6858C12Q2600/106C12Q2600/156C12Q2600/166C12Q2531/113C12Q2545/101C12Q2535/122C12Q2537/165Y02A50/30
Inventor 王子杰郑明顾民谭若芸居小兵韩志坚陶俊鲁佩陈浩孙黎费爽王增军
Owner JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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