Method for detecting gene polymorphism of UGT1A1 and liquid phase chip

Abstract

The invention discloses a liquid phase chip for detecting gene polymorphism of UGT1A1 and a detection method using the liquid phase chip, the liquid phase chip comprises microspheres respectively enveloped with specific corresponding wide-type and variant-type anti-tag sequences, wherein the anti-tag sequences are selected from the group consisting of SEQ ID NO.13 and SEQ ID NO.14 directed to UGT1A1*28 gene type, SEQ ID NO.9 and SEQ ID NO. 10 directed to UGT1A1*6 gene type, and / or SEQ ID NO.11 and SEQ ID NO.12 directed to UGT1A1*93 gene type; each of the above microspheres includes different color codings, primers including variant target sequences of the UGT1A1*28gene type, the UGT1A1*6 gene type and / or the UGT1A1*93 gene type are amplified, and the primers are modified by biotin so thatthe corresponding PCR reaction product contains a biotin labeling and has a sequence capable of complementary pairing with anti-tag. The matching rate of the detection method provided by the inventionand sequencing method reaches as high as 100%. The prepared liquid phase chip for detecting gene polymorphism of UGT1A1 has quite excellent signal-to-noise ratio and, basically, no cross reaction ispresent between the designed probe and the anti-tag sequence.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to medicine and biotechnology, in particular to a liquid phase chip for detecting UGT1A1 gene polymorphism and a detection method thereof. Background technique [0002] Irinotecan (CPT-11) is a chemotherapeutic drug that inhibits DNA topoisomerase I and is approved for marketing by the US FDA in 1998. During DNA replication, topoisomerase I reversibly cleaves single-strand DNA and reassembles to form double-strand DNA. The active metabolite SN-38 of irinotecan binds to the topoisomerase IDNA complex, prevents the reassembly of DNA strands, causes DNA double-strand breaks, and causes cell death. [0003] However, the decrease of neutrophils and severe enterotoxicity caused by irinotecan have become one of the key factors limiting its dosage. The active metabolite SN-38 of irinotecan is inactivated by hepatic uridine diphosphate glucuronosyltransferase (UDP-GT) (mainly metabolized by U...

AI Technical Summary

Problems solved by technology

Products currently on the market are mainly based on PCR-based detection technologies, such as direct sequencing, PCR-single-strand conformation polymorphism (SSCP) detection, these technologies have low sensitivity, easy sample contamination, and high false positive rate. Disadvantages, ordinary PCR method and fluorescent quantitative PCR cannot meet the clinical needs due to the limitation of detection throughput

However, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis technology and the allelic difference analysis method based on TaqMan technology can only detect one mutation at a time, which is time-consuming and laborious.

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