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AZF region microdeletion detection kit for Y chromosome

A Y chromosome and detection reagent technology, applied in the field of detection of microdeletions in the AZF region of the Y chromosome, can solve the problems of complex detection signal analysis, low accuracy and sensitivity, and high cost

Active Publication Date: 2015-12-23
PEKING JABREHOO MED TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Like the chip hybridization technology, this technology faces the defect of complex detection signal analysis. In addition, because the probe needs to be identified according to the length of the product, it has certain limitations in probe design and identification.
[0008] In summary, the current detection methods for microdeletions in the AZF region of the Y chromosome have the main defects of low accuracy and sensitivity, high cost, and complicated interpretation of detection results, etc.

Method used

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  • AZF region microdeletion detection kit for Y chromosome
  • AZF region microdeletion detection kit for Y chromosome
  • AZF region microdeletion detection kit for Y chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Hair follicle sample detection of microdeletions in AZF region

[0059] 1. Materials:

[0060] 1. Specimen: The source of the sample is a hair follicle sample from a male patient with oligospermia admitted to a cooperative hospital. The hospital used multiplex PCR technology to detect the deletion of the AZFc region of the Y chromosome.

[0061] 2. Reagents: Micro Genomic DNA Extraction Kit, End Filling Kit, dsDNAHS analysis kit, biotin, labeling enzyme, labeling enzyme reaction solution, ultrapure water, hybridization buffer, hybridization primer, DNA lysis solution (LowTE), ligase, ligase reaction solution, PCR reaction solution, label adapter, washing solution, XP magnetic beads, isopropanol, absolute ethanol, MyOneTMStreptavidinC1beads (MyoneC1 magnetic beads), 1MNaOH, Nuclease-freeWater (notDEPC-treated), IonPGMTTemplateOT2200Kit.

[0062] 3. Equipment: high-speed refrigerated centrifuge, water bath, ultrasonic interrupter CovarisM220, magnetic stand...

Embodiment 2

[0098] Example 2 Detection of microdeletions in AZF region in peripheral blood samples

[0099] 1. Materials:

[0100] 1. Specimen: The source of the sample is a peripheral blood sample from a male patient with oligospermia admitted to a cooperative hospital. The hospital used multiplex PCR technology to detect a partial deletion of the AZFc region of the Y chromosome.

[0101] 2. Reagents: plasma DNA extraction kit, dsDNAHS analysis kit, biotin, labeling enzyme, labeling enzyme reaction solution, ultrapure water, hybridization buffer, hybridization primer, DNA lysis solution (LowTE), ligase, ligase reaction solution, PCR reaction solution, label adapter, washing solution, XP magnetic beads, isopropanol, absolute ethanol, MyOneTMStreptavidinC1beads (MyoneC1 magnetic beads), 1MNaOH, Nuclease-freeWater (notDEPC-treated), IonPGMTTemplateOT2200Kit.

[0102] 3. Equipment: high-speed refrigerated centrifuge, water bath, magnetic frame, PCR instrument, IonOneTouch TM 2Instrumen...

Embodiment 3

[0135] Example 3 Detection of microdeletions in AZF region in peripheral blood samples

[0136] 1. Materials:

[0137] 1. Specimen: The source of the sample is the peripheral blood sample of a male patient with oligospermia admitted to the cooperative hospital. The hospital used multiplex PCR technology to detect that the AZF region of the Y chromosome was normal.

[0138] 2. Reagents: plasma DNA extraction kit, dsDNAHS analysis kit, biotin, labeling enzyme, labeling enzyme reaction solution, ultrapure water, hybridization buffer, hybridization primer, DNA lysis solution (LowTE), ligase, ligase reaction solution, PCR reaction solution, label adapter, washing solution, XP magnetic beads, isopropanol, absolute ethanol, MyOneTMStreptavidinC1beads (MyoneC1 magnetic beads), 1MNaOH, Nuclease-freeWater (notDEPC-treated), IonPGMTTemplateOT2200Kit.

[0139] 3. Equipment: high-speed refrigerated centrifuge, water bath, magnetic frame, PCR instrument, IonOneTouch TM 2Instrument, Io...

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Abstract

The invention provides a detection reagent for AZF (azoospermia factor) region microdeletion of a Y chromosome. The detection reagent comprises a primer composition, wherein primer sequences are designed for such primer sites aiming at the AZF region of the Y chromosome and other specificity regions serving as internal references; the characteristics of the primer sequences comprises: (1) in each group of primers, three continuous primers are designed by taking the previous sequence of a genome as a template, and form a PCR template after being connected; (2) a primer site sequence has a known and unchanged copy number in the target region of the chromosome of a normal person, and the sequence does not exist in other chromosome regions except the target region; (3) the sequence primers have close annealing temperatures, and the fluctuation range is lower than 5 DEG C; (4) the primer sequences do not have complementarity to a general amplification primer sequence; (5) an internal reference site only appears in the target region, and does not exist in other regions of the genome. The invention further provides a kit for detecting the AZF region microdeletion of the Y chromosome and a method for detecting the AZF region microdeletion of the Y chromosome.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit for detecting microdeletions in the AZF region of Y chromosome by using gDNA or free peripheral blood DNA and a method for using the kit to detect microdeletions in the AZF region of Y chromosome in vitro. Background technique [0002] A research report pointed out that infertile couples accounted for 15% of married couples of childbearing age, of which male factors accounted for about half, while 35% of male infertility was caused by genetic factors. Y chromosome microdeletion is the most important genetic factor of male infertility except K-type syndrome. While maintaining the evolutionary stability of the Y chromosome, the large number of repetitive sequences and palindromic structures of the Y chromosome also make the Y chromosome more prone to structural rearrangements, resulting in partial deletion or duplication of chromosomes. Y chromosome microdeletions ma...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6874C12Q1/6883C12Q2600/156C12Q2600/166C12Q2563/143C12Q2535/122C12Q2545/113
Inventor 冯涛李崎马燕琳李玉帅刘沙沙沈珺瑶
Owner PEKING JABREHOO MED TECH CO LTD
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