Method for detecting mitochondrial mutations and kit thereof

A technology of mitochondria and kits, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high cost, long cycle, cumbersome operation, etc., and achieve the effects of avoiding pollution, short time, and simple operation

Active Publication Date: 2011-08-17
智海生物工程(北京)股份有限公司
View PDF3 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For the detection of this mutation, in addition to the direct sequencing method, there are enzyme digestion sequence analysis and Taqman probe method, but these methods are not only expensive, but also cumbersome and long-term. To solve these problems, we designed a simple, fast, Inexpensive assays for clinical testing

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for detecting mitochondrial mutations and kit thereof
  • Method for detecting mitochondrial mutations and kit thereof
  • Method for detecting mitochondrial mutations and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0023] 1. Primer design

[0024] 1.1 Using the human mitochondrial genome nucleic acid sequence (NCBI database sequence NC_012920) as a template, design 1555A->G and 1494C->T mutant primers, so that the 3' ends of the primers are completely matched with 1555G and 1494T mutants, but with wild-type If there is no match, design an upstream primer shared with these two primers at the same time.

[0025] 1.2 In order to distinguish the Tm values ​​of the two amplified products on the melting curve, a sequence with high GC content was added at the 5' of the primer at position 1494.

[0026] 1.3 In order to ensure the validity of each detection, a pair of quality control primers was designed, and at the same time, it was ensured that the Tm value of the product could be distinguished from the other two detection peaks on the melting curve.

[0027] 1494 downstream primer: GCGGGCAGGGCGGCCTTTGAAGTATACTTGAGGAGA (Sequence 1), located between 1494-1514pb of the nucleic acid sequence, the...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for detecting mitochondrial mutations and a kit thereof. The method is used for detecting 1494T and 1555G mutations, five primers are shown as a sequence one, a sequence two, a sequence three, a sequence four and a sequence five in a primer sequence table, the primers are used for multiple polymerase chain reaction (PCR) amplification and melting curve analysis, and whether corresponding mutations happen is judged according to a melting peak at the specific temperature. Meanwhile, the invention also provides a kit special for the method.

Description

technical field [0001] The invention relates to the detection of gene mutation, more specifically, the detection of a mutation of human mitochondrial ribosomal RNA A1555G and C1494T related to drug-induced deafness. technical background [0002] Aminoglycosides such as streptomycin, gentamicin, kanamycin, sisomicin, tobramycin, amikacin, and netilmicin are particularly effective against Gram-negative bacterial infections. It is effective and has a synergistic antibacterial effect on some Gram-positive bacteria, and has become one of the most widely used antibiotics in clinical practice. However, this type of drug has serious ototoxic side effects, and may cause irreversible damage to hearing when used, even It is "one needle for deafness". [0003] This susceptibility is usually maternally inherited, which suggests that its possible pathogenesis is related to mitochondrial DNA (Mitochondrial DNA, mtDNA), and mutations in the mitochondrial 12S rRNA gene are one of the import...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 不公告发明人
Owner 智海生物工程(北京)股份有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap