Kit for directly detecting folic acid medication sensitive gene by use of blood and application method thereof

A kit and blood technology, applied in the field of medical testing, can solve the problems of nucleic acid loss and increased cross-contamination, and achieve the effect of promoting activity, avoiding cross-contamination, and intuitive judgment

Inactive Publication Date: 2017-08-18
浙江中迪生物科技有限公司
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  • Abstract
  • Description
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Problems solved by technology

[0007] The purpose of the present invention is to provide a kit for direct detection of folic acid drug-sensitive genes in blood with high amplification efficiency and low cost, aiming at solving the problem of prior art when performing fluorescent quantitativ

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  • Kit for directly detecting folic acid medication sensitive gene by use of blood and application method thereof
  • Kit for directly detecting folic acid medication sensitive gene by use of blood and application method thereof
  • Kit for directly detecting folic acid medication sensitive gene by use of blood and application method thereof

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Embodiment Construction

[0063] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. The specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0064] The embodiment of the present invention provides a kit for direct fluorescent PCR amplification of blood to detect human MTHFR gene polymorphism, including the following components: blood high tolerance taq polymerase, blood high tolerance taq polymerase special buffer, Blood samples, primer pairs, probe pairs, dNTPs, double distilled water;

[0065] The primers are MTHFRC677T primers and MTHFRA1298C primers,

[0066] The MTHFRC677T primers include:

[0067] Forward primer: MTHFRC677T-F: 5'-GCACTTGAAGGAGAAGGTGTCT-3';

[0068] Reverse primer: MTHFRC677T-R: 5'-TGTGTCAGCCTCAAAAGAAAAGCT-...

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Abstract

The inventiondiscloses a kit for directly detecting a folic acid medication sensitive gene by use of blood and an application method thereof. Human blood is adopted as a template, and a real-time fluorescence quantitative PCR system is formed by adding corresponding specific feature primers and fluorescent probes for detecting polymorphism of MTHFR C677T and MTHFR A1298C into the human blood respectively and amplifying a blood resistance taqpolymerase buffer and a blood resistance taqpolymerase. Through a Real time PCR reaction, a person skilled in the art determines the genotype of MTHFR according to the data collected by a fluorescence quantitative PCR instrument. The invention has the advantages of unnecessity of nucleic acid extraction, high speed and time saving, high sensitivity and easy judgment, low cost and the like, and is suitable for clinical detection and promotion.

Description

technical field [0001] The invention relates to the field of medical detection, and more specifically relates to a kit for directly detecting folic acid drug-sensitive genes in blood and a method for using the same. Background technique [0002] In recent years, with the development of biology, medicine, forensic science and other related fields, fluorescent PCR technology has been more and more widely used. As an important biological analysis sample, blood contains a large amount of DNA polymerase inhibitory substances, such as heme, miscellaneous protein, fat, etc. in blood. Therefore, based on the prior art, the fluorescent PCR method cannot directly amplify the nucleic acid in the blood. Usually, in order to complete the fluorescent PCR amplification of the nucleic acid of the blood sample, the nucleic acid substance must be purified from the blood first. It generally includes the following three processes: one is to rupture the cell membrane and nuclear membrane; the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q1/6858C12Q2600/106C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 林芳珍王晓洁
Owner 浙江中迪生物科技有限公司
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