Probe combination and kit for detecting microdeletion and microrepetition of Y chromosome and application of probe combination and kit

A Y-chromosome and microdeletion technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of undetectable fragment duplication, etc., and achieve intuitive and accurate structural interpretation, high sensitivity, and low The effect of human error

Pending Publication Date: 2022-02-08
王晴雪
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem that the current kits mentioned in the above background technology can only detect the deletion of chromosome segments, but cannot detect segment duplication, the present invention provides a probe combination for detecti

Method used

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  • Probe combination and kit for detecting microdeletion and microrepetition of Y chromosome and application of probe combination and kit
  • Probe combination and kit for detecting microdeletion and microrepetition of Y chromosome and application of probe combination and kit
  • Probe combination and kit for detecting microdeletion and microrepetition of Y chromosome and application of probe combination and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Embodiment 1-design of probe combination

[0079] A set of specific probes are designed for each detection point as shown in Table 1, and the length of the amplification product corresponding to each detection point is unique (88-168nt):

[0080] Table 1

[0081]

[0082]

[0083] The left sequence of the universal primer used by the above probe is shown in SEQ ID NO.51, and the right sequence is shown in SEQ ID NO.52.

Embodiment 2

[0084] Embodiment 2, sample preparation, PCR amplification program and result interpretation

[0085] A. Obtaining DNA samples to be tested

[0086] Select known normal married childbearing male samples, known normal female samples, known AZFb region deletion samples, known AZFc region deletion samples, known AZFb+c deletion samples, known AZFa+b+ c missing sample, DNA is extracted as the DNA sample to be tested in the present invention, and the pure water sample without DNA is used as a control;

[0087] B. The above-mentioned experimental materials are detected by the following detection methods, and the specific operations are as follows:

[0088] S100. Prepare the probe mixture: prepare 3 ul of the 25 groups of probe combinations in Example 1 with TE buffer solution, the final concentration of each probe is 1uM, and add the reference probe, the final concentration is 1uM;

[0089] Preparation of DNA template: prepare 100ng / 5ul DNA sample to be tested with TE buffer, heat...

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Abstract

The invention relates to the field of chromosome variation detection, in particular to a probe combination and a kit for detecting microdeletion and microrepetition of a Y chromosome and application of the probe combination and the kit, and the probe combination comprises specific probes aiming at 25 detection points (6 basic points and 19 extension points) of the Y chromosome. When the probe combination provided by the invention is adopted for detection, only Y chromosome microdeletion and microrepetition, 25 point locations, variation interval range judgment and related gene judgment are needed to be simultaneously detected aiming at DNA to be detected in the same reaction tube, and a qualitative and quantitative method containing 6 basic point locations and 19 expansion point locations is adopted. The technical means can be applied to various specimens such as semen, peripheral blood, tissues and the like, and the sample size is required to be only 20-50ngDNA (Deoxyribose Nucleic Acid); the method is short in detection time, economical, simple and convenient, capable of reducing manual operation errors, suitable for various types of samples, visual and accurate in structure interpretation and easy to popularize.

Description

technical field [0001] The invention relates to the field of chromosome variation detection, in particular to a probe combination, a kit and an application thereof for detecting Y chromosome microdeletion and microduplication. Background technique [0002] Y chromosome microdeletions are the second most common genetic cause of male spermatogenesis disorders. Associated chromosomal microdeletions are concentrated on three specific regions on the long arm of the Y chromosome (eg, figure 1 shown), respectively AZFa (15.2-13.1Mb), AZFb (22.8-18.5Mb) and AZFc (25.5-23.3Mb), deletions can affect single or multiple subregions, resulting in different clinical phenotypes. Recommended by the European Academy of Andrology (EAA) and the European Molecular Genetics Quality Network (EMQN) as a routine inspection item for men with azoospermia and severe oligospermia, the sY84 and sY86 sites in the AZFa subregion, and the sY127 and sY134 in the AZFb subregion site, as well as the sY254 an...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6827C12N15/11
CPCC12Q1/6888C12Q1/6827C12Q2531/113C12Q2565/125
Inventor 王晴雪
Owner 王晴雪
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