Detection kit for fluorescent PCR (Polymerase Chain Reaction) melting curve of gene mutation of glucose-6-phosphate dehydrogenase

Abstract

The invention discloses a detection kit for a fluorescent PCR (Polymerase Chain Reaction) melting curve of gene mutation of glucose-6-phosphate dehydrogenase. The kit provided by the invention can detect 16 mutations at 16 sites in a two-tube PCR system, and the genotype of a sample can be known through a primary fluorescent PCR melting curve analysis after PCR amplification. The whole operation is completed within 2-3 hours, so that the operating step is less, the time consumed is short, the flux is high, the detection sites are more and the mutation coverage is high. In addition, the kit provided by the invention uses homogeneous detection in a closed tube, so that the probability of PCR product pollution is reduced, the detection specificity is high and the result is easy to judge.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit for detecting the mutation of the glucose-6-phosphate dehydrogenase gene by using fluorescent PCR melting curve analysis. Background technique [0002] Glucose-6-phosphate dehydrogenase deficiency (G6PD deficiency) is one of the most common human enzyme deficiency diseases, and its cause is that G6PD gene mutations lead to decreased synthesis of glucose-6-phosphate dehydrogenase. However, the quality of life of the vast majority of patients has not been reduced. This is mainly because the vast majority of patients with G6PD deficiency in my country will have corresponding clinical symptoms only in the presence of incentives. And receive the correct treatment in time to effectively avoid the occurrence of the above situation. Therefore, it is very necessary to carry out universal screening in areas with a high incidence of G6PD deficiency in my country. [0003] The...

AI Technical Summary

Problems solved by technology

[0004] Although the biochemical detection method has low detection cost and short detection time, it has the following disadvantages: (1) the detection rate of female heterozygotes is low, only about 20-45%; (2) the results are easily affected by the experimental conditions, operators and other factors

In addition, because some point mutation types have little change in the spatial conformation of single-stranded DNA, it is easy to miss detection

Gene chip technology is not only costly and complicated to operate, but also results in varying degrees of false positives

Images