Kit for fluorescent quantitative PCR detection of EML4-ALK fusion gene

Abstract

The invention provides a kit for fluorescent quantitative PCR detection of an EML4-ALK fusion gene. The kit comprises a reaction buffer solution, MgCl2, dNTP, a primer mixed solution I, a primer mixedsolution II, a primer mixed solution III, a primer mixed solution IV, a primer mixed solution V, a primer mixed solution VI, a primer mixed solution VII, a primer mixed solution VIII, DNA polymerase,purified water and a positive control, the concentration of each of above primers and probes is 10 [mu]mol, and a volume ratio of a primer to a probe in every mixed solution is 2.5:1. A amplificationreaction system of the kit includes 0.2 [mu]l of the DNA polymerase, 1.2 [mu]l of the primer mixed solution, 2 [mu]l of a sample / positive control / purified water, 12.1 [mu]l of purified water, 2 [mu]lof MgCl2, 2 [mu]l of 10 * buffer and 0.5 [mu]l of the dNTP. The kit has a high detection sensitivity and a good specificity; the specific primers and fluorescent probes are designed for seven variants of the EML4-ALK fusion gene, and a relevant reaction solution is detected in tubes to eliminate the interference among the primers, so the detection specificity and the sensitivity in the inventionare greatly higher than those of a case adopting a mixed reaction solution, and the false positive rate is low.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting EML4-ALK fusion gene fluorescence quantitative PCR. Background technique [0002] Lung cancer is a malignant tumor with the highest morbidity and mortality in the world. Over the past 50 years, the incidence and mortality of lung cancer have risen rapidly in countries all over the world, especially in industrialized countries. In 1995, 600,000 people died of lung cancer in the world, and the number is increasing every year. In 2003, the World Health Organization (WHO) announced that the mortality rate was 1.1 million / year, and the incidence rate was 1.2 million / year. The etiology of lung cancer is still not completely clear so far. Medical statistics show that the pathogenic factor of lung cancer is mainly smoking (including second-hand smoke). 80% of male lung cancer and 75% of female lung cancer are caused by smoking. In addition, it is associated with asbestos...

AI Technical Summary

Problems solved by technology

Although FISH is a sensitive and specific method for the detection of ALK fusion genes in lung adenocarcinoma, it is not foolproof and cannot differentiate between different EML4-ALK fusion gene variants

There is still no uniform standard for the treatment strategy and result interpretation of different numbers of fracture signals