Probe, kit and method thereof for detecting MAPT gene mutation

Abstract

The invention discloses a probe, a kit and a method thereof for detecting MAPT gene mutation, and relates to the field of bio-medicine. The probe for detecting the MAPT gene mutation includes, but not limited in, at least one of a TaqMan probe, a TaqMan-MGB probe, a molecular beacon, a modified molecular beacon, a double-stranded fluorescent displacing probe, a LightCycler probe, a double-loop probe and the like. The kit for detecting the MAPT gene mutation consists of an amplification primer, a fluorescent probe, an amplification reagent and a reference reagent. The method for detecting the MAPT gene mutation comprises the following steps: 1) selecting a sample; 2) extracting genome DNA; 3) conducting real-time fluorescent PCR amplification and detection; and 4) conducting result analysis. Experimental results can be observed in real time. Meanwhile, the probe, the kit and the method are more sensitive and rapid in detection, more convenient to operate and easy for automation, and the probability of cross pollution can be greatly reduced and the accuracy of detection can be improved. Cost is saved, and meanwhile flux is improved.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a probe, a kit and a method for detecting MAPT gene mutation. Background technique [0002] Progressive supranuclear palsy (PSP), also known as Steele-Richardson-Olszewski syndrome, is a clinically rare degenerative disease of the central nervous system. PSP is the most common degenerative parkinsonian syndrome after Parkinson's disease (PD), and its incidence accounts for about 6%-10% of PD. It was first discovered and described by Posey in 1904. In 1964, Steele et al. (Steele JC, Richardson JC, et al. .Arch Neurol,1964,10:333-359.) This disease is listed as an independent nervous system disease. The main clinical features of PSP are postural instability, vertical supranuclear palsy, pseudobulbar palsy, supranuclear oculomotor disturbance, cervical dystonia, dysarthria, pseudobulbar palsy, mild dementia and paresthesia. Similar symptoms of Kinson syndrome, etc. [0003] The onset ...

AI Technical Summary

Problems solved by technology

But there are many deficiencies in this method: first, this method relies on the activity of restriction endonucleases, and it is easy to produce false positive and false negative results in the process of enzyme digestion; secondly, the method of electrophoresis is likely to cause contamination of PCR products, resulting in erroneous results. Again, this method is time-consuming (about 5-6 hours) and labor-intensive, including at least four steps of PCR amplification, enzyme digestion, electrophoresis, and gel imaging analysis, requiring a lot of manual operations, low degree of automation, and large human errors

At present, there is no report on the use of real-time fluorescent PCR to study the mutations of MAPT genes R5L, A152T, L284R, S285R, N296del and G303V

Images