Liquid chip for detection of fshr gene mutation and its detection method

A detection solution and chip technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the limitation of detection flux that cannot meet clinical needs, poor repeatability of detection results, and expensive solid-phase chips and other problems, to achieve accurate and reliable detection results, improve detection accuracy, and improve the fluorescence signal value

Inactive Publication Date: 2011-12-07
SUREXAM BIO TECH
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Problems solved by technology

[0005] Several PCR-based technologies for detecting FSHR SNPs have been established, such as direct sequencing, semi-quantitative PCR, and PCR-single-strand conformational polymorphism (SSCP) detection. The above technologies have low sensitivity, easy contamination of samples, and Disadvantages such as high false positive rate, ordinary PCR method and fluorescent quantitative PCR cannot meet the clinical needs due to the limitation of detection throughput
However, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis technology and the allelic difference analysis method based on TaqMan technology can only detect one mutation at a time, which is time-consuming and laborious; traditional solid-phase chips Expensive, low sensitivity and poor reproducibility of test results

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  • Liquid chip for detection of fshr gene mutation and its detection method
  • Liquid chip for detection of fshr gene mutation and its detection method
  • Liquid chip for detection of fshr gene mutation and its detection method

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Embodiment 1

[0029] FSHR gene mutation detection liquid chip kit mainly includes:

[0030] 1. ASPE Primers

[0031] The wild-type and mutant-specific primer sequences were designed for the five types of mutations on exon 10 of FSHR. ASPE primers are composed of Tag+ specific primer sequence. The principle of specific primer design is: the 3' end of the specific primer is the mutation site; the specific primers used to detect the wild type and mutant type of a gene mutation site should come from DNA The same strand; the annealing temperature of the specific primer should be between 51°C and 56°C; the length of the primer is generally between 18-22bp. The ASPE sequence is shown in the table below:

[0032]Table 1 ASPE primer sequences

[0033]

[0034] Each ASPE primer includes two parts, the 5' end is a specific tag sequence for the anti-tag sequence on the corresponding microsphere (as shown in Table 2), and the specific tag sequence is complementary to the corresponding anti-tag seq...

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Abstract

The invention discloses a liquid phase chip for FSHR gene mutation detection and a detection method thereof. The liquid phase chip includes: microspheres respectively coated with specific anti-tag tag sequences; and mutant-specific ASPE primers; primers that amplify the target sequence with the mutation site on exon 10 of the FSHR gene. The coincidence rate of the detection method provided by the present invention and the sequencing method is as high as 100%, and the prepared FSHR gene mutation detection liquid chip has a very good signal-to-noise ratio, and the designed probe and anti-tag sequence are basically There is no cross-reactivity.

Description

technical field [0001] The invention relates to medical in vitro diagnostic technology, in particular to a liquid phase chip for FSHR gene mutation and a detection method thereof. Background technique [0002] Follicular stimulating hormone receptor (FSHR) is a member of the glycoprotein subfamily of the G protein-coupled receptor superfamily, and consists of three parts: extracellular region, intracellular region and transmembrane region. The specific FSHR on the membrane surface of ovarian granulosa cells binds to follicle stimulating hormone (FSH), and plays an important role in promoting the development of follicles in the ovary in women. After the combination of FSH and FSHR, by activating the FSHR promoter, activating RNA polymerase to accurately combine with the template DNA and start gene transcription, causing changes in the post-transcriptional level of the FSHR gene, thereby mediating the response signal of gonadotropin, leading to the first Two messengers produc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 许嘉森朱泽尧陈玲郭元杰
Owner SUREXAM BIO TECH
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