Specific primers and liquid phase chip for polymorphic detection of human hedgehog interacting protein (HHIP) gene

Abstract

The invention discloses specific primers and a liquid phase chip for the polymorphic detection of a human hedgehog interacting protein (HHIP) gene. The liquid phase chip comprises allele specific primer extension (ASPE) primers consisting of tag sequences at the 5' end and specific primers at the 3' end, microspheres which are wrapped with specific anti-tag sequences and an amplification primer, wherein the sequences of the specific primers are selected from more than one pair of SEQ ID No. 5 and SEQ ID No. 6 aiming at T149C and SEQ ID No. 7 and SEQ ID No. 8 aiming at T61C. The liquid phase chip for the polymorphic detection of the HHIP gene has a high signal noise ratio, and cross reaction is not formed between a designed probe and the anti-tag sequences basically; and by the selection of the tag sequences and the anti-tag sequences and the combination of the tag sequences and the specific ASPE primers, the cross reaction can be prevented, and the parallel detection of multiple single nucleotide polymorphism (SNP) loci can be realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for HHIP gene polymorphism detection and a liquid phase chip. Background technique [0002] Hedgehog (Hh) signaling pathway is a signaling pathway that plays an important role in animal embryonic development and cell proliferation. It has been found that the abnormal activation of this pathway plays an important role in the occurrence of various tumors. The human hedgehog interacting protein (Human Hedgehog interacting Protein, HHIP) gene is located on chromosome 4q31.21-31.3, and a group of transmembrane glycoproteins encoded by this gene can combine with the three Hh proteins in the Hedgehog signaling pathway, thereby negatively Regulates Hedgehog signaling pathway activity. Current studies have shown that polymorphisms of the HHIP gene are associated with the risk of various diseases such as breast cancer, chronic lung ...

AI Technical Summary

Problems solved by technology

[0005] At present, there are almost no products on the market to detect HHIP gene polymorphism at home and abroad, and there are very few reports on the gene polymorphism, and most of the reports are still in the experimental research stage and have not yet been commercialized. The existing detection technologies are mainly based on On the basis of PCR technology, such as direct sequencing method, real-time fluorescent quantitative PCR method, etc., these technologies have the disadvantages of easy sample contamination and high false positive rate, and at the same time, due to the limitation of detection throughput, they cannot meet the clinical needs

Again, these methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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