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ABCB1 and CYP3A5 gene detection primer group, kit and detection method

A gene detection and primer set technology, applied in the fields of genetic engineering and molecular biology, can solve the problems of low detection sensitivity, high false positive rate, and long cycle, and achieve high throughput, reduce false positive rate, and short detection cycle Effect

Inactive Publication Date: 2019-03-08
WUHAN CONSIDERIN GENE & HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a kind of ABCB1, CYP3A5 gene detection primer set, kit and detection method, aim at solving the low sensitivity of ABCB1, CYP3A5 gene detection in the prior art, high false positive rate, long period, low throughput technology question

Method used

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  • ABCB1 and CYP3A5 gene detection primer group, kit and detection method

Examples

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no. 1 example

[0033]The present invention proposes a first embodiment, a primer set for detection of ABCB1 and CYP3A5 genes, which is used to specifically amplify nucleic acid fragments containing sites to be tested, and the sites to be tested include rs1045642 and rs1045642 on the ABCB1 gene The rs776746 site on the CYP3A5 gene, the primer set includes the rs1045642 primer set and the rs776746 primer;

[0034] The rs1045642 primer set includes rs1045642 outer primer set and rs1045642 inner primer set, the rs1045642 outer primer set includes rs1045642 outer upstream primer SEQ ID NO:1 and rs1045642 outer downstream primer SEQ ID NO:3; the rs1045642 inner primer set includes The upstream primer SEQ ID NO:2 in rs1045642 and the downstream primer SEQ ID NO:4 in rs1045642;

[0035] The rs776746 primer set includes rs776746 outer primer set and rs776746 inner primer set, the rs776746 outer primer set includes rs776746 outer upstream primer SEQ ID NO:5 and rs776746 outer downstream primer SEQ ID ...

no. 2 example

[0037] The present invention proposes a second embodiment, an ABCB1 and CYP3A5 gene detection kit, which contains the ABCB1 and CYP3A5 gene detection primer set of the first embodiment above.

[0038] Preferably, the kit further includes an adapter containing a tag sequence, and the adapter is used for direct connection with the amplification product containing rs1045642 and rs776746. The tag sequence is nnnn, wherein n is A, G, C or T. Adapters of the invention can take a variety of forms including, but not limited to, blunt-ended adapters, protruding-end adapters.

[0039] Preferably, the linker sequence also includes a sequence complementary to the sequence of the sequencing primer and the sequence of the amplification primer.

[0040] Preferably, the kit further includes an enzyme that specifically cleaves uracil, and the enzyme that specifically cleaves uracil is USER enzyme, RNase H or UDG enzyme.

[0041] Preferably, the linker is a protruding end linker. Since the se...

no. 3 example

[0051] The present invention proposes a third embodiment, a method for detecting ABCB1 and CYP3A5 genes, comprising the following steps:

[0052] A. Use the primer set to amplify the samples containing the rs1045642 site and the rs776746 site respectively to obtain amplified products; the primer set includes the rs1045642 primer set and the rs776746 primer;

[0053] The rs1045642 primer set includes rs1045642 outer primer set and rs1045642 inner primer set, the rs1045642 outer primer set includes rs1045642 outer upstream primer SEQ ID NO:1 and rs1045642 outer downstream primer SEQ ID NO:3; the rs1045642 inner primer set includes The upstream primer SEQ ID NO:2 in rs1045642 and the downstream primer SEQ ID NO:4 in rs1045642;

[0054] The rs776746 primer set includes rs776746 outer primer set and rs776746 inner primer set, the rs776746 outer primer set includes rs776746 outer upstream primer SEQ ID NO:5 and rs776746 outer downstream primer SEQ ID NO:7; the rs776746 inner primer ...

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Abstract

The invention relates to the field of genetic engineering and molecular biology and provides an ABCB1 and CYP3A5 gene detection primer group which is used for specific amplification of a nucleic acidsegment containing to-be-detected sites including rs1045642 site on ABCB1 gene and rs776746 site on CYP3A5 gene. The primer group comprises an rs1045642 primer and an rs776746 primer. The invention further provides a gene detection kit and a detection method. The gene detection primer group can realize specific amplification of the nucleic acid segment containing the to-be-detected sites, therebybeing high in sensitivity and low in false positive rate in ABCB1 and CYP3A5 gene mutation detection; second-generation high-throughput gene sequencing technology is adopted to detect genotype of theto-be-detected sites, so that detection is short in period and high in throughput.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, and more specifically relates to a primer set, a kit and a detection method for detecting ABCB1 and CYP3A5 genes. Background technique [0002] Paclitaxel is a tricyclic diterpenoid compound with a unique anti-tumor mechanism. Its broad anti-cancer spectrum and definite clinical efficacy have attracted people's attention. Later research found that paclitaxel is a specific stabilizer of microtubules, which can promote the assembly of microtubules and maintain the stability of microtubules. When the cells are in contact with paclitaxel, a large number of microtubules will accumulate in the cells, and the accumulation of these microtubules will interfere with various normal functions of the cells, especially to stop cell division at the mitotic stage and block the normal division of cells. Based on the above mechanism, paclitaxel is the only herbal drug that can control the...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q2531/113C12Q2535/122C12Q2525/191
Inventor 盛司潼徐云符兆威
Owner WUHAN CONSIDERIN GENE & HEALTH TECH CO LTD
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