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Primer combination and kit used for detecting Y chromosome microdeletion

A chromosomal microdeletion and detection kit technology, which is applied in the field of biotechnology detection, can solve the problems of increasing workload and analysis difficulty, increasing the complexity of result interpretation, and failing to improve detection sensitivity, and achieves intuitive result interpretation and short detection time. , the effect of saving testing costs

Active Publication Date: 2017-04-26
北京圣谷智汇医学检验所有限公司
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AI Technical Summary

Problems solved by technology

In 2013, EAA and EMQN published a revised version of the guidelines for the molecular diagnosis of Y chromosome microdeletions, pointing out the method of using the above 6 STS loci to detect the deletion of the AZF region (usually two multiplex PCR reactions, each reaction to detect different regions 3 STS loci) can effectively and accurately diagnose Y chromosome microdeletions, but new detection methods that add or include more STS loci cannot improve the sensitivity of detection, and may even increase the complexity of interpretation of the results, so It is not recommended to add more STS sites
Two amplification reactions not only increase the workload and difficulty of analysis, but also increase the chance of cross-contamination due to sample confusion or other reasons

Method used

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  • Primer combination and kit used for detecting Y chromosome microdeletion
  • Primer combination and kit used for detecting Y chromosome microdeletion
  • Primer combination and kit used for detecting Y chromosome microdeletion

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Embodiment Construction

[0079] The term "Y chromosome microdeletion" as used herein refers to the deletion of a small segment of a gene on the Y chromosome. In the present invention, the Y chromosome microdeletion is mainly the deletion of the azoospermia factor (AZF) region, and different degrees of deletion of the AZF region can lead to azoospermia or oligospermia.

[0080] The term "azoospermia factor (AZF)" used herein refers to a gene that causes azoospermia and is located on the long arm of the Y chromosome. Usually, AZF is divided into three regions, AZFa, AZFb and AZFc, in which AZFa is located in the 5th region of the long arm of the Y chromosome, AZFb is located in the 5-6 region of the long arm of the Y chromosome, and AZFc is adjacent to the heterochromatin region. Such as figure 1 shown. Deletion of the AZF gene results in oligospermia or azoospermia.

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Abstract

The invention relates to a primer combination and kit for chromosome microdeletion. By the use of a multiple PCR combined with agarose gel electrophoresis technology, seven STS loci closely associated with the Y chromosome microdeletion are detected to judge the microdeletion conditioof a Y chromosome. Seven STS loci with high specificity and sensitivity (sY84, sY86, sY127, sY134, sY254, sY255 and sY160) are chosen as detection loci, at the same time two loci ZFX / Y and SRY are utilized as quality control loci, a primer is designed aiming at each locus respectively, and products of the nine loci after multiplex amplification are directly put on the agarose gel to be conducted electrophoresis detection. By the use of the primer combination and detection system, the Y chromosome microdeletion can be detected only needing single tube amplification reaction.

Description

technical field [0001] The invention relates to the field of biotechnology detection, in particular to a primer combination and a kit for detecting Y chromosome microdeletion. Background technique [0002] It is estimated that approximately 15% of couples of reproductive age are infertile, with male factors accounting for approximately 50%. The main cause of male infertility is the man's spermatogenesis disorder, manifested as severe oligospermia (less than 2×10 6 / ml~5×10 6 / ml) or no sperm, and about 20% of clinically infertile men are caused by hereditary, non-obstructive azoospermia or oligospermia, and most of these patients have normal karyotype tests. Among the known genetic factors leading to male infertility, Y chromosome microdeletion is the second most common genetic factor after Klinefelter syndrome. [0003] In 1976, Tiepolo et al., in analyzing the karyotypes of more than 1,000 male infertility patients, found that the long arm of the Y chromosome in several...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6806C12Q1/686C12Q1/6883C12Q2600/118C12Q2600/156C12Q2600/16C12Q2521/101C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 周仲春叶建伟张捷邱丽丽刘棒余应襄崔丹
Owner 北京圣谷智汇医学检验所有限公司
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