Rapid detection kit for Hcy metabolic key enzyme and detection method

A technology for detecting kits and key enzymes, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of Hcy serum level deviation, interference test results, work impact, etc., to achieve accurate results and simplify the test process , good accuracy

Pending Publication Date: 2018-10-09
新开源云扬(广州)医疗科技有限公司
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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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  • Rapid detection kit for Hcy metabolic key enzyme and detection method
  • Rapid detection kit for Hcy metabolic key enzyme and detection method
  • Rapid detection kit for Hcy metabolic key enzyme and detection method

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Embodiment Construction

[0038] A rapid detection kit and detection method for a key enzyme in Hcy metabolism provided by the present invention will be further described in detail and completely in conjunction with the examples below. The embodiments described below are exemplary only for explaining the present invention and should not be construed as limiting the present invention.

[0039] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples were purchased from the market unless otherwise specified.

[0040] The instruments and reagents adopted in the present invention are as follows:

[0041] PTC-255 PCR amplification instrument, MJ Research, USA;

[0042] Pyrosequencer, Wuhan First Biotechnology Co., Ltd.;

[0043] TaqDNA polymerase, dNTPs (10mmol / L), 10×Buffer and MgCl 2 (25mmol / L) (Dalian TaKaRa company); AmpliTaqGold polymerase (AppliedBiosystems company); KlenowFragment (no exonucle...

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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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2531/113C12Q2535/122C12Q2565/301
Inventor 王花孙力强孙正扬孙正明
Owner 新开源云扬(广州)医疗科技有限公司
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