Kit for detecting gene mutation of aminoglycoside drugs

An aminoglycoside and kit technology, used in genomics, microbial determination/inspection, bioinformatics, etc., can solve the problems of limited number of samples tested, high sequencing cost, difficulty in interpretation of results, etc., to achieve low cost, sustainable The effect of high sensitivity and specificity, easy to popularize and apply

Pending Publication Date: 2021-11-12
苏州源河医疗科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

From the perspective of patents, PCR-based methods are the main ones. CN107881229A, CN112852948A and CN111172250A have announced the method of designing primers based on PCR to detect these two mutations. It is indeed easy to operate, high in efficiency and low in cost, but the interpretation results are easily affected by sample concentration and purity. and other factors, and the number of detection samples is limited by the detection channel
The Sanger method is a low-throughput technique that can only dete

Method used

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  • Kit for detecting gene mutation of aminoglycoside drugs
  • Kit for detecting gene mutation of aminoglycoside drugs
  • Kit for detecting gene mutation of aminoglycoside drugs

Examples

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Embodiment 1

[0046] A kit for detecting gene mutations of aminoglycoside drugs according to this embodiment, the kit includes the probes shown in SEQ ID No.1~SEQ ID No.2.

[0047] The design of the probe is combined with the literature reports of clinical laboratories, aiming at the clinical diagnosis and treatment guidelines related to aminoglycoside drug genes, combined with databases such as ClinVar, CPIC and pharmaGKB to screen these sites, and confirmed 2 high-incidence sites in the Chinese population , and synthesize the corresponding probes for these two sites and make a chip, with one probe for each site. The screening principles are as follows:

[0048] (1) High-incidence sites in the Chinese population included in clinical drug guidelines;

[0049] (2) The ClinVar database is rated as Drug response, and the evidence strength is 3 stars or above;

[0050] (3) The CPIC and pharmaGKB databases publish Guidelines and the CPIC grade is A.

[0051] The components of the kit also inc...

Embodiment 2

[0056] In the present embodiment, clinical samples are actually detected by the present invention, which specifically includes the following steps:

[0057] (1) Sample processing and quality control: 2 clinically obtained samples were used as positive verification samples for the invention. These verification samples are first subjected to DNA extraction, and quality control is carried out through gel electrophoresis, Nanodrop, etc. to ensure that each DNA sample has no degradation, no impurity contamination, and high purity. The optical density (Optical Density, OD) 260 / 280nm ratio is between 1.8~2.0, and the OD 260 / 230nm ratio is between 1.5~2.0. Samples that do not meet any of the conditions need to be purified and other treatments. The gel loading dye is Invitrogen™ TrackIt Cyan / Orange Loading Buffer (Invitrogen P / N 10482-028); 25bp Invitrogen Ladder (Invitrogen P / N 10488-022); the gel electrophoresis system is Invitrogen E-Gel™ 48 agarose gels 4%, G8008-04.

[0058] (2)...

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Abstract

The invention provides a kit for detecting gene mutation of aminoglycoside drugs. The kit comprises two hotspot variations m.1494C exceeding T and m.1555A exceeding G of a mitochondrial gene MT-RNR1. The kit can detect m.1494C exceeding T and m.1555A exceeding G accurately, quickly, efficiently and conveniently at low cost, adopts a microarray chip technology, continues high sensitivity and specificity of a traditional gene chip, is easy to popularize and apply clinically, can quickly screen m.1494C exceeding T and m.1555A exceeding G in a neonatal stage and effectively prevents one-shot deafness of children.

Description

technical field [0001] The invention belongs to the field of gene detection, in particular to a kit for detecting gene mutations of aminoglycoside drugs. Background technique [0002] Aminoglycoside drugs include gentamicin, streptomycin, kanamycin, etc., which are mainly used clinically to treat infections caused by Gram-negative bacteria and Mycobacterium tuberculosis infections. The combination of 16S rRNA leads to mistranslation of bacterial protein or premature termination of protein synthesis to achieve antibacterial effect. Studies have shown that in some children carrying 12S rRNA gene m.1494C>T and m.1555A>G mutations, the secondary structure of 12S rRNA undergoes the same changes as bacterial 16S rRNA. When it is combined with 12S rRNA in the human body, the mitochondrial protein synthesis of the ear cells is abnormal, reducing the amount of ATP synthesis, further making it impossible to maintain the ion gradient of the inner ear, resulting in an imbalance o...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G16B20/50G06K9/62
CPCC12Q1/6883G16B20/50C12Q2600/156C12Q2600/106G06F18/23
Inventor 黄守锋
Owner 苏州源河医疗科技有限公司
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