Multiplex PCR specific primers, kit and method for detecting causative genes of large vestibular aqueduct syndrome based on high throughput sequencing technique

Abstract

The invention belongs to the field of gene detection and relates to multiplex PCR specific primers, kit and method for detecting causative genes of large vestibular aqueduct syndrome based on a high throughput sequencing technique. The invention discloses the multiplex PCR specific primers which are used for detecting causative genes of large vestibular aqueduct syndrome and are shown in formulas of SEQ ID NO: 1 to SEQ ID NO: 86, the kit containing the primers and the detection method for capturing and amplifying the causative gene of large vestibular aqueduct syndrome through a one-step PCR amplification reaction through the primers or kit. The multiplex PCR specific primers, kit and detection method can realizes simultaneous detection of multiple causative genes of large vestibular aqueduct syndrome of multiple samples, effectively improve the detection efficiency and accuracy, reduce a cost and simplify operation processes.

Description

technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a multiplex PCR specific primer, kit and method for detecting the causative gene of large vestibular aqueduct syndrome based on high-throughput sequencing technology. Background technique [0002] Large vestibular aqueduct syndrome (Large Vestibular aqueduct Syndrome, LVAS) is a congenital hereditary inner ear deformity hearing impairment discovered in the late 1970s. A characteristic autosomal recessive genetic hearing impairment disease, clinically mainly manifested as sensorineural deafness with high-frequency hearing loss, and the degree of hearing loss is usually severe or extremely severe deafness, which may be accompanied by A series of clinical symptoms such as recurrent tinnitus or vertigo. The onset is mostly in childhood, and before the onset, there are often colds, fevers, traumas and other causes that increase intracranial pressure. A number of clinical st...

AI Technical Summary

Problems solved by technology

Sanger sequencing can only sequence a certain region of a sample at a time, which has low detection efficiency, high cost, and low sensitivity (20%)

Gene chip method and fluorescent quantitative PCR method are used to detect the pathogenic gene of large vestibular aqueduct syndrome. This method can only design detection probes for one or several known mutation sites to form a detection chip, so it cannot be used to detect unknown mutation sites. point, the test results are not comprehensive and accurate enough

Multi-color fluorescent melting curve analysis method, the fluorescent dye used is a general-purpose double-stranded DNA intercalator, which cannot recognize primer dimers and non-specific amplification products, and may easily cause false positives

The high-throughput sequencing method is mainly composed of sequencing instrument library construction kits provided by sequencing instrument suppliers, which are evaluated by the supplier company for quality control. There are shortcomings such as long delivery period of kits, fixed specifications of kits, and insufficient test flexibility.

Some reagent manufacturers build libraries according to the experimental methods shared and reported in the literature, but the results of the completed experiments are uneven, and the sequencing quality is not high

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