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Kit for detecting microdeletion of human Y chromosome

A Y chromosome and kit technology, which is applied in the field of kits for detecting microdeletions of human Y chromosomes, can solve problems such as unfavorable promotion, high price of sequencers and HPLC, and high professional requirements for result interpretation, so as to avoid false negatives, The effect of reducing the risk of contamination and reducing the workload

Active Publication Date: 2014-12-24
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sequencers and HPLC are expensive, and require high professionalism in instrument operation and result interpretation, which is not conducive to promotion

Method used

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  • Kit for detecting microdeletion of human Y chromosome
  • Kit for detecting microdeletion of human Y chromosome
  • Kit for detecting microdeletion of human Y chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Technical basis

[0035] 1.1 Use the known Y chromosome microdeletion and STS research results to design and implement the multiplex PCR reaction solution.

[0036] Its main research contents include: according to the requirements of the 1999, 2004 and 2013 editions of the EAA / EMQN guidelines and the recommended STS, design different fluorescently labeled STS detection probes; according to the characteristics of STS in different regions and the probe labeling group, respectively Three tubes of PCR reaction system were designed, and each tube of reaction system was used for multiple fluorescent PCR amplification with corresponding STS amplification primers and probes to specifically detect different STSs. The human zinc finger protein gene ZFX / Y was used as an internal control in each reaction system, and the male sex-determining gene (Sex-determining Region of Y—Chromosome, SRY) was simultaneously amplified in the first reaction system as a sex abnormal control.

[...

Embodiment 2

[0058] Embodiment 2 The use of kit of the present invention

[0059] 1. Background of clinical indications:

[0060] The kit described in Example 1 of the present invention is used for the qualitative detection in vitro of microdeletions in the region of the azoospermia factor (AZF) affecting spermatogenesis on the human Y chromosome. Including the detection of 6 sequence-tagged sites (STS) closely related to the deletion of small DNA fragments in the three regions of AZF (AZFa, AZFb, AZFc), namely sY84, sY86, sY127, sY134, sY254, sY255.

[0061] This kit is only used for the detection of microdeletion genes in the AZF region of the Y chromosome. It can be used as a molecular inspection method to find the cause of male infertility. The results cannot be directly used as the final diagnosis basis for male infertility.

[0062] 2. Inspection principle

[0063] This kit product uses multiplex PCR combined with multicolor Taqman fluorescent probe technology.

[0064] Multiplex ...

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Abstract

The invention relates to the field of chromosome deletion, in particular to a kit for detecting the microdeletion of a human Y chromosome. The kit comprises PCR (Polymerase Chain Reaction) reaction liquid I, PCR reaction liquid II and PCR reaction liquid III, wherein the PCR reaction liquid I comprises primers and fluorescent probes of specific amplification ZFX / Y, sY254, sY134 and SRY (Sex-determine Region of Y Chromosome) sites; the PCR reaction liquid II comprises primers and fluorescent probes of specific amplification ZFX / Y, sY84 and sY127 sites; the PCR reaction liquid III comprises primers and fluorescent probes of specific amplification ZFX / Y, sY255 and sY86 sites. According to the kit disclosed by the invention, by designing the specific fluorescent probes of different STS (Sequence Tagged Site) and three-pipe reaction systems are optimally combined, so that microdeletion types of the Y chromosome can be detected by one-time test; effective monitoring for different reaction systems is realized by the ZFX / Y.

Description

technical field [0001] The invention relates to the field of chromosome deletion detection, in particular to a kit for detecting human Y chromosome microdeletion. Background technique [0002] Azoospermia factor (AZF), which is closely related to spermatogenesis, exists on the long arm of the human Y chromosome. The small deletion of DNA fragments in the azoospermia factor is called Y chromosome microdeletion. The occurrence of Y chromosome microdeletion can cause spermatogenesis disorder, resulting in oligozoospermia, weak sperm, teratozoospermia or even azoospermia. According to WHO statistics, about 10%-15% of male patients with primary azoospermia and oligospermia have Y chromosome microdeletions. In male infertility patients caused by spermatogenesis disorder, the incidence of Y chromosome microdeletion is the second genetic factor. [0003] At present, AZF is divided into 3 regions, namely AZFa, AZFb and AZFc. Any one or more deletions in these regions will lead to s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2537/143C12Q2561/101C12Q2561/113
Inventor 滕乐林斯里田洁
Owner 亚能生物技术(深圳)有限公司
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