Rapid detection kit for Hcy metabolic key enzyme and detection method

Abstract

The invention discloses a rapid detection kit for a Hcy metabolic key enzyme and a detection method. The kit consists of an efficient amplification group and a pyrophosphoric acid sequencing group, wherein the efficient amplification group includes a specific amplification primer; a template sequence to be amplified is included in a sequence shown as SEQ ID NO:1; the efficient amplification groupis used for linear exponential PCR (Polymerase Chain Reaction) amplification of a template to be amplified; an amplification product is sequenced by the pyrophosphoric acid sequencing group after being annealed; the pyrophosphoric acid detection group includes an annealing primer, DNA (Deoxyribonucleic Acid) polymerase, ATP (Adenosine Triphosphate) sulfurylase, luciferase and a Klenow enzyme. A target fragment amplification method of linear exponential amplification is adopted, a precisely-designed amplification primer has high amplification efficiency and few mismatches, directly-amplified single-stranded DNA can be directly applied to pyrophosphoric acid sequencing without labeling and double chain separation of the amplification product, tedious experimental steps are avoided, and damage to amplified fragments by alkaline reagents is also reduced.

Description

technical field [0001] The invention relates to a rapid detection kit and a detection method for key enzymes of Hcy metabolism, belonging to the field of biological sample detection. Background technique [0002] Homocysteine ​​(Hcy) is a sulfur-containing amino acid in the human body. It is an intermediate product in the metabolic process of methionine and cysteine. It itself does not participate in protein synthesis and cannot be directly synthesized in the body. Hcy can only be transformed from methionine in food after demethylation. In recent years, a large number of studies have shown that hyperhomocysteinemia (hyperhomocysteinemia, HHcy) is another major pathogenic factor leading to cardiovascular and cerebrovascular diseases, elevated plasma homocysteine ​​and people with hypertension It has a synergistic effect on causing cardiovascular and cerebrovascular diseases. The relationship between hyperhomocysteinemia and some diseases such as cerebrovascular disease, cor...

AI Technical Summary

Problems solved by technology

Therefore, when the serum level of Hcy is used as the judgment standard for many diseases, if the change in serum level caused by the mutation of key enzymes is not considered, it may lead to a large deviation in the serum level of Hcy, which will interfere with the judgment of the test results. This has a great impact on subsequent work

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