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Amplification composition and kit for detection of y chromosome microdeletion

A Y chromosome and microdeletion technology, applied in the field of amplification compositions and kits for Y chromosome microdeletion detection, can solve problems such as increasing the difficulty of primer design, and achieve the effects of intuitive and accurate result interpretation, short detection time, and simple operation.

Active Publication Date: 2022-07-29
BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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Problems solved by technology

[0004] Multiplex PCR combined with agarose gel electrophoresis is currently the most widely used method for Y chromosome microdeletions. After years of clinical practice, this method has been proved to be accurate, sensitive, easy to operate, and low in cost. However, the biggest difficulty and challenge of this method lies in the multiple Design of PCR primers: (1) Accurately locate the position of the STS and design primers to avoid STS loss; (2) Design the primers scientifically and reasonably to avoid mutual interference; (3) The size difference between the amplified fragments must reach more than 50bp, agar Glycogel electrophoresis can be used to better distinguish and interpret the results
These limitations greatly increase the difficulty of primer design

Method used

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  • Amplification composition and kit for detection of y chromosome microdeletion
  • Amplification composition and kit for detection of y chromosome microdeletion
  • Amplification composition and kit for detection of y chromosome microdeletion

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Embodiment Construction

[0063] A kit for detection of Y chromosome microdeletion is detected by the method of PCR amplification-agarose gel electrophoresis, comprising the following steps:

[0064] (1) Design of primer combinations

[0065] According to the recommendation of EAA / EMQN guidelines, 6 STS sites (AZFa: sY84, sY86; AZFb: sY127, sY134; AZFc: sY254, sY255) and 2 quality control sites (ZFX / ZFY, SRY) were selected and searched by NCBI Design and synthesize the best amplification primers based on the DNA sequence of each site to ensure that the size gap between each amplified fragment is more than 77bp, and will not interact to produce non-specific products or dimers. Then, dilute to the required concentration of 10 μM as needed. The specific primer sequences are shown in Table 1, as shown in SEQ ID NOs: 1-16 in the sequence table:

[0066] Table 1 Primer sequences

[0067]

[0068] (2) DNA extraction

[0069] The collected fresh peripheral blood samples were used to extract genomic DNA u...

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Abstract

The invention discloses an amplification composition and a kit for detecting Y chromosome microdeletion, which are used to amplify 6 STS sites (AZFa: sY84, sY86; AZFb: sY127, sY134; AZFc: sY254, sY255) and The primers of 2 quality control sites (ZFX / ZFY, SRY) were detected by multiplex PCR combined with agarose gel electrophoresis technology to detect the above-mentioned 6 STS sites that are closely related to Y chromosome microdeletion, so as to judge the Y chromosome microdeletion. Happening. Using the amplification composition of the present invention, only 2 tubes of amplification reaction can be used to simultaneously detect 6 STS sites.

Description

technical field [0001] The invention relates to the technical field of chromosome deletion detection, in particular to an amplification composition and a kit for Y chromosome microdeletion detection. Background technique [0002] The azoospermic factor (AZF) region that affects spermatogenesis exists on the long arm of the Y chromosome, which can be further divided into three regions: AZFa, AZFb and AZFc. Microdeletions in either region can lead to spermatogenesis disorders in men, leading to oligospermia or azoospermia. Among male infertility patients caused by spermatogenesis disorders, the incidence of Y chromosome microdeletion is second only to Klinefelter's syndrome, which is the second most common genetic factor. Y chromosome microdeletion has become a routine examination item for male infertility patients. [0003] Among the three regions of AZF, each region has a clear sequence tag site (STS), and the position of the deletion break has been clear. Considering tha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6883C12Q1/6858C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 智慧芳佟洪梅赵方圆倪君君
Owner BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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