c-KIT somatic mutation gene detecting kit and detecting method thereof

Abstract

The invention relates to a c-KIT somatic mutation gene detecting kit and a detecting method thereof. The kit comprises primers and probes for detecting c-KIT somatic cell gene mutation. The primers and the probes are specifically the primer and the probe for detecting a No. 11 exon of a c-KIT gene, wherein a primer sequence is as shown in SEQ ID NO: 1-6, and a probe sequence is as shown in SEQ IDNO: 7-8; the primer and the probe are used for detecting a No. 9 exon of the c-KIT gene, wherein the primer sequence is as shown in SEQ ID NO: 9-10, and the probe sequence is as shown in SEQ ID NO: 11; the primer and the probe are used for detecting a No. 13 exon of the c-KIT gene, wherein the primer sequence is as shown in SEQ ID NO: 12-14, and the probe sequence is as shown in SEQ ID NO: 15; andthe primer and the probe are used for detecting a No. 17 exon of the c-KIT gene, wherein the primer sequence is as shown in SEQ ID NO: 16-19, and the probe sequence is as shown in SEQ ID NO: 20. Thedetecting kit and the detecting method are far more than similar products in detecting site number, complete in site, high in detecting sensitivity, strong in specificity, high in accuracy, low in cost, and capable of meeting clinical GIST targeted administration c-KIT mutation detection.

Description

technical field [0001] The invention relates to nucleic acid in vitro amplification detection in the field of biotechnology, in particular to a c-KIT somatic cell mutation gene detection kit and a detection method thereof. Background technique [0002] Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor of the digestive tract, and it is also a model in tumor biological targeted therapy. The vast majority of GISTs express the Kit protein (CD117) encoded by the c-KIT gene. The c-KIT gene is located on human chromosome 4q12-13 and belongs to a proto-oncogene. Some GISTs have c-KIT gene mutations, which lead to the activation of kit protein without the participation of ligands to stimulate the continuous proliferation of tumor cells and the loss of control of anti-apoptotic signals. [0003] Imatinib (Gleevec) is a tyrosine kinase inhibitor approved by the FDA for the treatment of metastatic and surgically refractory cases, as well as postoperative preven...

AI Technical Summary

Problems solved by technology

Among them, the Sanger sequencing method has high accuracy as the gold standard, but its sensitivity is low, it takes a long time, and it is difficult to be used in clinical detection; the HRM method detection has higher requirements on the performance of the instrument, and the sensitivity is not high

In addition, several companies use fluorescent PCR method to detect c-KIT mutations, but the detection sites are limited to 1-2. The reason is that blocking probes or locked nucleic acid modifications are added to the detection reagents, the detection cost is high, and the number of detection sites is far away. Less than the number of clinically required tests, so the clinical application is limited

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