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Composition and product for detecting Y chromosome microdeletion, application of composition and method

A technology of Y chromosome and composition, applied in the field of molecular biology, can solve the problems of operation and instrument requirements, complicated operation, difficult to popularize, etc., and achieve the effects of accurate results, high sensitivity and simple operation.

Pending Publication Date: 2020-10-16
SHANDONG SHANDA HOSPITAL FOR REPRODIVE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method has extremely high requirements on operation and instruments, and the operation is extremely cumbersome, so it is difficult to popularize in practical applications and is not conducive to popularization.

Method used

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  • Composition and product for detecting Y chromosome microdeletion, application of composition and method
  • Composition and product for detecting Y chromosome microdeletion, application of composition and method
  • Composition and product for detecting Y chromosome microdeletion, application of composition and method

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0047] Adopting positive quality control product and negative quality control product, product of the present invention is compared with a market product (Y chromosome microdeletion detection kit (PCR fluorescent probe method) produced by Shanghai Toujing Life Science and Technology Co., Ltd.), wherein , the positive quality control product is identified as normal male genomic DNA (no deletion at each site), and the negative quality control product is deionized water. The use method of the product of the present invention is the same as the method introduced in this manual, and the use of the product on the market is carried out according to the instructions of the kit. The result is as Figure 1-4 shown.

[0048] The fluorescence intensity values ​​at each point detected by the product of the present invention and the products on sale in the market are compared, and the results are as shown in Table 1:

[0049] Table 1: Product of the present invention compares with the tes...

experiment example 2

[0053] Select 46 examples (S1-S46) of AZF detection in the Affiliated Reproductive Hospital of Shandong University to carry out the verification test, adopt the test kit of the present invention and the commercially available product test kit used in Experimental Example 1 to detect respectively, and the two results are inconsistent Samples were then verified by gel electrophoresis. Wherein the use method of the kit of the present invention is as described herein, and the use of the product kit on the market is carried out according to the contents of the kit instructions. The gel electrophoresis method is as follows:

[0054] The following primers and PCR Mix (recipe as above) were prepared as premix A (ZFX / SRY / SY86 / SY127 / SY153 primers), B (ZFX / SRY / SY254 / SY134 / SY255 primers), C (RBM1 / SRY / SY84 / SY152 primer):

[0055]

[0056] Remarks: The gel electrophoresis method includes three spots of RBM1 (region b), SY152 (region d), and SY153 (region d), and the present invention ...

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Abstract

The invention relates to the field of molecular biology, and provides composition for detecting Y chromosome microdeletion. The composition comprises primers and probes with sequences as shown in SEQID NO: 1 to SEQ ID NO: 24. The invention also provides a product comprising the composition and a method using the composition. The composition has the characteristics of simple operation, short time,less pollution, accurate result and the like.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a composition, product and application for detecting Y chromosome microdeletion. Background technique [0002] The Y chromosome regulates the growth and development of male sperm cells and plays an important role in male fertility. Therefore, the damage of Y chromosome function is likely to lead to a variety of phenomena. [0003] Y chromosome microdeletion refers to the occurrence of DNA fragments or gene deletions in functional domains on the Y chromosome. The Y chromosome microdeletion region mainly includes AZFa, AZFb, AZFbc, AZFc, and AZFabc regions ("2013 New EAA / EMQNY Chromosomal Microdeletion Detection Guidelines Interpretation"). The "Practice Guidelines for the Molecular Diagnosis of Y Chromosome Microdeletions" jointly issued by the European Academy of Andrology (EAA) and the European Molecular Genetics Quality Network (EMQN) recommended multiplex PCR detection of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6883C12Q1/6858C12Q2600/156C12Q2600/16C12Q2531/113C12Q2561/101C12Q2537/143
Inventor 高媛康冉冉黄色新徐佩文邹洋李杰高明马金龙陈子江
Owner SHANDONG SHANDA HOSPITAL FOR REPRODIVE MEDICINE
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