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Kit for detecting regulon of Leber disease and use thereof

A technology of regulators and kits, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of increased false negatives and strict PCR amplification conditions, and achieves low cost, intuitive result interpretation, and detection process. easy effect

Inactive Publication Date: 2012-10-24
ZHEJIANG UNIV
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Problems solved by technology

Although this method can detect positive sites in a short period of time, the PCR amplification conditions are strict, and the probability of false negatives is greatly increased

Method used

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  • Kit for detecting regulon of Leber disease and use thereof
  • Kit for detecting regulon of Leber disease and use thereof
  • Kit for detecting regulon of Leber disease and use thereof

Examples

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Effect test

Embodiment 1

[0036] Example 1 Detection of Leber hereditary optic neuropathy family carrying mitochondrial gene A4435G mutation and primary G11778A mutation, see figure 1 .

[0037] 1. Test samples

[0038] A family with Leber's hereditary optic neuropathy (WZL4 family) carrying the A4435G mutation was selected. See his pedigree figure 2 . This family presents a typical maternal inheritance, and the only clinical symptom of the patients is uncorrectable visual impairment, but the degree of visual impairment of each member of the family ranges from mild to severe. The total number of people in this family is 25, including 14 members of the maternal line, and 8 people with vision loss.

[0039] 2. Extraction of Genomic DNA

[0040] Obtain the blood filter paper pieces (one drop of blood) of these 8 subjects respectively, and cut the blood filter paper pieces into about 1cm in size with clean scissors 2 Put the shredded pieces of paper into a 1.5ml EP tube (Eppendorf tube), add 900μl...

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Abstract

The invention provides a kit for detecting mitochondrial DNA mutation related to a regulon of a Leber disease, wherein the kit consists of primer sequences, reagents for extracting genome DNA of samples, reagents for PCR amplification reaction, a positive control sample, a negative control sample and an instruction, restrictive endonucleases comprise a restrictive endonuclease Nla III, and the positive control sample is a restriction enzyme digested sample in which 4435th site of a mitochondrial sequence occur A to G mutation. The kit provided by the invention can be applied in the detection of mitochondrial DNA mt DNAA4435G mutation related to the regulon of the Leber disease. According to the kit, specificity and stability of detection results can be ensured, cost is lower than that of other detection methods, and the detection process is simple, quick, accurate and economic, and thus is suitable for detecting LHON-related gene mutation.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a method for detecting mitochondrial gene mutations related to regulators of Leber disease, in particular to detecting mitochondrial tRNA Met The method for the A4435G mutation also relates to a kit for detecting the mitochondrial DNA A4435G mutation related to the Leber disease regulator, and the application of the above method or the kit in detecting the mitochondrial DNA A4435G mutation related to the Leber disease regulator. Background technique [0002] Leber's Hereditary Optic Neuropathy (Leber's Hereditary Optic Neuropathy, LHON, referred to as Leber's disease) is a type of optic neuropathy, mainly involving the retina and the anterior part of the cribriform plate of the sclera, the papillary macular tract fibers, a maternally inherited disease that causes optic nerve degeneration, severe It affects people's normal production and life. According to the latest statistic...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 管敏鑫冀延春蒋萍萍张娟娟肖云
Owner ZHEJIANG UNIV
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