Kit for specifically detecting genotype of ApoE4 and application of kit

Abstract

The invention relates to a kit for specifically detecting the genotype of ApoE4 and application of the kit. The kit comprises sucrose water of 4 percent, a NaOH solution of 10mmol/L, a Tris-HCL (pH 7.5) solution of 1mol/L, DMSO (Dimethylsulfoxide), a reagent I, a reagent II, Hha I incision enzyme and 10*M Buffer. The application of the kit comprises the following steps of: preparing and preserving a sample; extracting genome DNA; carrying out PCR (Polymerase Chain Reaction) amplification; and identifying the genotype of ApoE4. Comparing with the prior art, the reagent provided by the kit can noninvasively obtain individual genome DNA; the ApoE4 gene can be efficiently amplified, the genotype of the ApoE4 can be quickly and accurately determined, and the genotype determination of a large quantity of individuals can be simultaneously carried out; and the kit has the advantages of short determination time and high accuracy and is suitable for large-scale screening of molecular epidemiology and preventive medicine.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a rapid and specific test kit for detecting ApoE4 genotype and its application. Background technique [0002] Human apolipoprotein E (Apolipoprotein E, ApoE) is encoded by 19q11.2, and its gene commonly has three phenotypes, namely three apolipoprotein E2, E3 and E4 encoded by ε2, ε3 and ε4 alleles , forming six different genotypes. Including 3 types of homozygotes (ε2 / ε2, ε3 / ε3, ε4 / ε4) and 3 types of heterozygotes (ε2 / ε3, ε3 / ε4, ε2 / ε4). Allelic changes cause ApoE4 to have greater affinity for LDL receptors, that is, ApoE4>ApoE3>ApoE2, thereby affecting lipoprotein metabolism and blood lipid levels. ApoE4 / 4 genotype has been used as an independent risk factor for cardiovascular and cerebrovascular diseases. Since the ApoE gene is related to insulin resistance (IR), obesity, etc., the detection of ApoE genotype is of great significance for the study of medical molecular ep...

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Problems solved by technology

However, this method requires high sampling and cumbersome detection, which is not conducive to the use of epidemiological census, and invasive blood drawing to obtain genomic DNA is not accepted by every individual, especially the

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