Detection kit and detection method for calcium ion antagonist metabolism marker and application thereof

Abstract

The invention discloses a detection kit and a detection method for calcium ion antagonist metabolism marker and application thereof. The kit comprises the following components: a CYP3A5 * 3 amplification primer, a CYP3A5 * 3 sequencing primer, an NPPA (T2238C) amplification primer, an NPPA (T2238C) sequencing primer and a positive control. According to the invention, CYP3A5 * 3 and NPPA (T2238C) are amplified through multiple RPA in one tube, so that a large number of amplification products are rapidly and effectively generated at a constant temperature. Single-stranded DNA is specifically captured through amino-labeled single-stranded DNA directly combined with carboxyl modifiers, after washing, sequencing primers and sequencing raw materials are added to perform pyrosequencing, damage of strong alkaline reagents to amplified fragments is reduced, the sequencing process is simplified, and the sequencing time is shortened.

Description

technical field [0001] The invention relates to a detection kit for a calcium ion antagonist metabolic marker, a detection method and an application thereof, and belongs to the field of gene detection. Background technique [0002] At present, there are five categories of antihypertensive drugs commonly used in clinical practice: diuretics, angiotensin converting enzyme inhibitors, angiotensin receptor blockers, β-blockers, and calcium ion antagonists. Drug metabolism in the body involves a variety of drug-metabolizing enzymes, transporters, and receptors, and their genetic polymorphisms ultimately lead to the therapeutic effect, adverse development, and risk factors when patients take the same drug in the same way. There are obvious individual differences in drug tolerance and other aspects. [0003] Calcium antagonists are chemicals that lower blood pressure by blocking calcium channels. It can selectively inhibit Ca2+ from entering the cell through the calcium channel o...

AI Technical Summary

Problems solved by technology

Among them, the sequencing method and the chip method have cumbersome operation steps, long detection cycle, and the amplification product is prone to contamination; the high-resolution melting curve method has simple steps, low specificity, and high requirements for equipment; The specific amplification method uses ARMS primers for specific amplification, and the design of the primers is difficult to optimize, and the detection conditions are strict.

Taqman fluorescent probe method has high test cost, and the amplification throughput of multiple genes is not high

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