Method and kit for detecting mutation of PDS gene 2168A>G by using fluorescence PCR technology

Abstract

The invention provides a method and a kit for detecting mutation of PDS gene 2168A>G by using fluorescence PCR technology. The method comprises: aiming at a PDS 2168A>G mutation site, designing a specific mutation type probe and a wild type probe and respectively marking with different colors, designing amplimers at two sides of the mutation site; and performing amplification on a to-be detected sample by using a PCR reaction system containing the wild type probe, the mutation type probe and the amplimers, and comparing the fluorescence curve mode with that of a quality-control sample to determine whether the sample has PDS gene 2168A>G mutation. The method and the kit can provide clinical reference for disease cause diagnosis of deaf patient, especially deaf patients with enlarge vestibular aqueduct syndrome, also can be used to screen whether newborns and couples in pregnancy are carriers, and provides help for deaf antenatal diagnosis and newborn congenital deaf cause diagnosis. The kit has the characteristics of closed-tube high-flux automatic operation and analysis, and is especially usable in clinic laboratories.

Description

technical field [0001] The invention belongs to the technical field of gene detection, in particular to a method for detecting the mutation of deafness gene PDS 2168A>G by probe-specific real-time PCR technology and a kit thereof. Background technique [0002] Deafness is the most common disease causing language communication barriers, and the incidence of newborn deafness is 0.1% to 0.3%. There are many people with hearing and language disabilities in my country, and the number of deaf children born every year is increasing at a rate of 30,000. In my country, about 300,000 couples of childbearing age who have at least one deaf child face the choice of having another child. The cause of deafness, whether it is hereditary, and whether the next childbirth is safe are issues of great concern to deaf families. Enlarge vestibular aqueduct syndrome (EVAS) is an autosomal recessive hearing impairment disease with a relatively high incidence. Imaging examination data show that ...

AI Technical Summary

Problems solved by technology

Direct sequencing of PCR products is currently the most widely used and more accurate method, but this method requires expensive instruments and professional personnel to operate, time-consuming and laborious, and it is difficult to promote in clinical practice

The gene chip method has the advantages of fast and high throughput, but the operation is complicated, there are many steps, the cost is high and it is not easy to popularize

The method of detecting gene mutation with restriction fragment length polymorphism (RFLP) has the advantage of low cost, but the process is cumbersome and takes a long time and is not suitable for popularization

The published patent 201010599385.X uses specific primers and probes to detect mutations through single nucleotide extension technology combined with microarray chip technology. This method is complex in operation, expensive in instruments, and difficult to popularize

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