Detection method of hypertriglyceridemia mutant site and detection kit

Abstract

The invention discloses a detection method of a hypertriglyceridemia mutant site. Through the design of a specific primer, a target fragment is amplified by single-tube multiplex PCR (polymerase chainreaction); a sequencing primer is designed; a non-glue-cutting purified Sanger sequencing method is used for performing sequencing on the obtained target fragment. The invention also discloses a detection kit of the hypertriglyceridemia mutant site. The multiplex amplification LPL (lipoprotein lipase) isogene and the non-glue-cutting purification sequencing can be realized; only the target gene is amplified; the advantages of low cost and exact target goal are realized. Dozens of fragments are amplified at the same time through a single tube; the detection flux is high; the problem of time consumption and labor waste of the single PCR are solved; the operation is simple and fast; the operation can be performed by ordinary experiment personnel according to the description; during the sequencing, the glue cutting purification is not needed; the result shows that the operation is convenient; the PCR multiple number is sufficient; the identical or similar band strip size does not need tobe worried. The popularization is easy; the market prospects are wide.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a detection method and a detection kit for hypertriglyceridemia mutation sites. Background technique [0002] Lipoprotein lipase (LPL) is the key enzyme for the hydrolysis of triglycerides. The disorder of this enzyme can lead to the aggregation of plasma chylomicrons and lipoproteins rich in triglycerides. The clinical manifestations are chylomicronsemia, blood triglycerides Elevated esters. Studies have shown that LPL gene defects or mutations can lead to severe hypertriglyceridemia (HTG). In addition to LPL, gene mutations or defects such as LMF1, GPIHBP1, APOA5, and GPD1 can also cause severe primary hypertriglyceridemia. Variations / defects in the above genes can affect LPL activity, hinder the hydrolysis of triglycerides, and eventually lead to severe hypertriglyceridemia. For patients with gene mutations / defects, timely use of molecular screening, effective diagnosis and preci...

AI Technical Summary

Problems solved by technology

In order to reduce costs, whole exome sequencing and target sequence sequencing methods have been developed, and multiple samples are mixed together for sequencing to further reduce sequencing costs, such as the application of gene panels designed for cancer and other complex diseases, but The tedious sample preparation and pooling, library building, sequencing, assembly, analysis and other processes take a long time, which is not conducive to the rapid diagnosis and follow-up monitoring of diseases, which limits its clinical application, and is more used in clinical or biological academic research

In addition to sequencing technology, methods for genetic testing include qPCR technology, gene chip technology, etc., none of which can provide complete sequence information

qPCR can detect rare mutations but its throughput is low and only known mutations can be detected

The technical cost of the gene chip is high and the sensitivity is low

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