ATP7B (ATPase Cu2+transporting beta polypeptide) gene mutation detection primer set and kit, ATP7B gene mutation detection method and uses of ATP7B gene mutation detection primer kit

A technology for detecting primers and primer sets, applied in the field of molecular biology, can solve the problems of easy misdiagnosis and missed diagnosis, irreversible damage to patients, missed optimal treatment time, etc., and achieves the effect of uniform sequencing results, low cost and high sensitivity

CN106011302AInactive Publication Date: 2016-10-12深圳市人口和计划生育科学研究所
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2016-10-12
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to the field of biology and especially relates to an ATP7B (ATPase Cu2+transporting beta polypeptide) gene mutation detection primer set and kit, an ATP7B gene mutation detection method and uses of the ATP7B gene mutation detection primer kit. The primer set relates to the most area of the ATP7B gene, comprises an exon and an intron, acquires complete ATP7B gene nucleotide sequence cases and determines a mutation site. The primer set aims at long fragment amplification, a single stripe can be obtained through amplification under appropriate conditions, and a nonspecific stripe is not produced. The amplified fragment is long so that compared with the short fragment amplification method, the long fragment amplification method produces a uniform sequencing result and data analysis is simpler.
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Description

technical field

[0001] This application relates to the field of molecular biology, in particular to a primer set for detecting ATP7B gene mutations, a kit, a detection method and applications thereof. Background technique

[0002] Hepatolenticular degeneration, also known as Wilson Disease (WD), is a severe autosomal recessive disorder of copper metabolism with different ages of onset and different clinical phenotypes. According to foreign epidemiological survey data, the prevalence rate is 1.5-3.0 / 100,000, and the gene frequency is 0.3-0.7%. The carrier rate in the Chinese Han population is 1 / 50. The most common clinical manifestations of hepatolenticular degeneration are liver cirrhosis, neurological symptoms, and psychiatric symptoms. Hepatolenticular degeneration is mainly diagnosed through clinical manifestations such as liver and / or nervous system symptoms, family history, corneal pigment ring, biochemical examination including detection of serum ceruloplasmin, 24-ho...

Examples

Embodiment 1

[0082] 1. Family situation: The clinical manifestations of the proband were abnormal liver function, elevated transaminase, ceruloplasmin 69.6 mg / L, and corneal K-F ring (-). There were no other patients in the family of both parents. The parents of the proband had normal phenotypes, non-consanguineous marriage, and no history of drug use or adverse environmental exposure during pregnancy. This study was approved by the Medical Ethics Committee of the Shenzhen Institute of Population and Family Planning Sciences, and all genetic diagnoses were obtained with the consent of the patient's family members and signed informed consent.

[0083] 2. Method:

[0084](1) Specimen collection and DNA extraction: Take 2 mL of peripheral venous blood from the proband and his parents, and anticoagulate with sodium citrate. Whole blood genomic DNA was extracted using QIAamp DNA MiniKit (Qiagen) and stored at -20°C. (2) Analysis of the full-length gene sequence of the pathogenic gene: Screen ...