ApoE gene typing detection kit suitable for blood genome DNA sample and PCR method therefor

Abstract

The invention provides an ApoE gene typing detection kit suitable for a blood genome DNA sample and a PCR method therefor. The kit includes a mixed liquid used for ApoE gene typing detection and a quality control substance containing recombinant DNA; wherein the mixed liquid includes a specific primer pair used for the ApoE gene typing detection, a specific probe pair, Taq HS DNA polymerase, a deoxyribonucleoside triphosphate mixed liquid, a reaction buffer liquid, and a reaction enhancer. In the invention, by combining the basic principles of fluorescent PCR-MGB probe method and AMRS-PCR method, the enhancer is employed and the raw material ratio is optimized, so that detection sensitivity is increased. The kit is free of addition of a housekeeping gene or an internal reference, so that compared with similar technologies, the kit is convenient to use, is more rapid in detection, has higher accuracy, is especially suitable for detection on the blood genome DNA sample, and is proper topromotion and application and industrialization.

Description

technical field [0001] The kit related to the present invention belongs to the technical field of in vitro detection, and is a kit suitable for ApoE genotyping detection of blood genome DNA samples and a PCR method thereof. Background technique [0002] ApoE is the abbreviation of apolipoprotein E (apolipoprotein E). ApoE is mainly produced in the liver and brain. Its main function is to participate in the transportation of lipids, especially cholesterol, in the blood circulation system and central nervous system, and it is also a low-density lipid. Ligands of protein and very low-density lipoprotein and chylomicron receptors, and involved in the regulation of enzyme activities related to lipoprotein metabolism. The phenotype of ApoE is controlled by three alleles located on one gene, namely E2 , E3 and E4, each allele corresponds to a major isoform to produce three homozygotes (E2 / E2, E3 / E3, E4 / E4) and three heterozygotes (E2 / E3, E2 / E4, E3 / E4) a total of six common phenot...

AI Technical Summary

Problems solved by technology

[0004] (1) The ApoE gene involves two sites, and the multi-tube detection has many steps and is easy to form false positives. However, the single-tube multiplex PCR detection method is limited due to the lack of multiple fluorescence channels of many manufacturers’ fluorescent PCR instruments. Restrictions, result analysis is also more troublesome;

[0005] (2) The sequence of the ApoE gene is a typical sequence with high GC content. The detection sensitivity of ordinary fluorescent probes is relatively low. The concentration of DNA samples extracted with blood genomic DNA extraction kits is about 10-100ng / μL. The blood extraction rate is lower and contains PCR inhibitors such as heme, so there is a method using fluorescent PCR, the concentration of DNA extracted under special conditions is too low and contains impurities, and the DNA sample extracted from blood cannot be accurately detected classification

Images