Detection kit for CAG trinucleotide duplication mutation of SCA (Spinocerebellar Ataxia) pathogenic genes

Abstract

The invention performs genetic diagnosis on common SCA subtypes 1, 2, 3, 6, 7, 8, 17 and DRPLA of Chinese population by using a method of agarose gel electrophoresis, capillary electrophoresis and T carrier clone sequencing. Mutation detection on the CAG trinucleotide duplication mutation of the SCA pathogenic genes comprises three parts: firstly, performing the agarose gel electrophoresis on a DNA sample of a patient, and performing preliminary diagnosis; secondly, performing the capillary electrophoresis on the DNA sample of the patient to determine times of the CAG trinucleotide duplication mutation, and performing further diagnosis; thirdly, performing the T carrier clone sequencing on corresponding subtypes of the sample which is diagnosed to be sicken in the above steps, so as to determine internal structures of the pathogenic genes, and finally confirming diagnosis.

Description

technical field [0001] The present invention relates to the field of molecular biology and diagnostic medicine. More specifically, the present invention relates to genetic diagnosis of common SCA subtypes 1, 2, 3, 6, 7, 8, 17 and DRPLA in the Chinese population by means of agarose gel electrophoresis, capillary electrophoresis and T vector cloning and sequencing. Background technique [0002] Hereditary spinocerebellar ataxia (SCA) is a class of progressive neurodegenerative diseases including multiple subtypes of ataxia, with autosomal dominant inheritance, with an incidence of about 5 / 100,000, accounting for neurological deficits. 10%-15% of systemic hereditary diseases. The lesions are mainly located in the cerebellum, brain stem, and spinal cord, and the pathological changes are mainly in the form of neuronal cell loss and gliosis. The clinical manifestations are cerebellar ataxia, accompanied by dysarthria, intention tremor, eye movement disturbance, pyramidal tract a...

AI Technical Summary

Problems solved by technology

[0004] At present, in clinical application, the CAG trinucleotide repeat mutation detection of the SCA pathogenic gene is used for further molecular diagnosis of patients with clinically diagnosed SCA, and most of them only use a certain method for genetic diagnosis, among which polyacrylamide gel electrophoresis or agarose gel electrophoresis Glycogel electrophoresis has the advantage of high sensitivity, but its specificity is low, and the specific number of CAG repeats cannot be determined

Although capillary electrophoresis can determine the number of CAG repeats with a certain accuracy, it cannot determine the internal structure of the mutant gene well, and its diagnostic specificity and sensitivity are relatively low.

T vector clone sequencing can determine the internal structure of the gene well, but cannot accurately determine the number of CAG repeats due to possible DNA strand slippage

[0005] SCA1, 2, 3, 6, 7, 8, 17, DRPLA and other subtypes are the eight subtypes with the highest incidence of SCA in the Chinese population. The current diagnostic methods are mostly based on the analysis of individual genes one by one. Specific diagnostic kit for each subtype

Images