Combined detection kit for human tumor gene mutation

Abstract

The invention provides a combined detection kit for human tumor gene mutation. The combined detection kit comprises the following parts: an EGFR (Epidermal Growth Factor Receptor) gene primer group, a KRAS gene primer group, a BRAF (serine-threonine kinase) gene primer group, an NRAS gene primer group, a PIK3CA (Phosphatidylino-sitol 3-kinases) gene primer group, a HiFi enzyme, a PCR (Polymerase Chain Reaction) reaction solution, a digestive enzyme, ligase, a ligation buffering solution, a ligation joint, a fluorescent probe, a Taq enzyme of QPCR (Quantitative Polymerase Chain Reaction) reaction, a QPCR primer and a QPCR reaction solution. The combined detection kit provided by the invention can be used for simultaneously detecting mutant sites of a plurality of driving genes of human tumors, including one or more gene mutant sites of EGFR, NRAS, KRAS, PIK3CA and BRAF; an experiment result has high sensitivity and the mutation rate of 1 percent can be detected; the detection sensitivity is greatly improved and the difficulties in the prior art are overcome; meanwhile, requirements on large-batch and rapid detection are met through a whole experiment process.

Description

technical field [0001] The invention relates to the field of gene variation detection, in particular to a combined detection kit for human tumor gene variation. Background technique [0002] Lung cancer is a common cancer in the world, and its morbidity and mortality rank first among all cancers. In 2002, the new incidence of lung cancer in the world was 1.38 million, nearly half of which occurred in developing countries. According to statistics from the Ministry of Health of my country in 2008, the mortality rate of lung cancer was 30.83 / 100,000. Lung cancer has become the first malignant tumor in terms of morbidity and mortality, and the number of deaths from lung cancer is far greater than the sum of deaths from breast cancer and prostate cancer. [0003] Lung cancer can be divided into two categories based on histopathology: small cell lung cancer and non-small cell lung cancer, of which non-small cell lung cancer accounts for about 85% of all lung cancer cases, includ...

AI Technical Summary

Problems solved by technology

In the Sanger sequencing method, a single pair of primers can detect multiple mutations, but the mutation sites of multiple genes or different exons of the same gene need to be amplified and sequenced separately, the operation is cumbersome, and the sensitivity is about 20% lower , a higher false positive rate

Although the fluorescent quantitative PCR method has high sensitivity, it can only detect one mutation for each pair of primers, and each mutation requires a separate PCR system, and the operation of detecting multiple mutation sites at the same time is cumbersome

These two methods require a large amount of samples and are not suitable for detecting multiple gene mutation sites at the same time

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