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Real-time fluorescence quantification PCR (Polymerase Chain Reaction) kit and method for detecting Y-chromosome micro-deletion

A real-time fluorescence and Y-chromosome technology, applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of low detection throughput, high cost, inapplicability, etc., to reduce time and The effect of cost, high sensitivity and specificity, and high detection throughput

Active Publication Date: 2014-09-17
骆子义 +1
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Problems solved by technology

The latter is the gold standard for all methods but is not widely used due to its high cost
Although the PCR-RFLP method has low cost and simple operation, its specificity and sensitivity are not ideal, and its detection throughput is small, so it is not suitable for clinical detection with a large sample size.

Method used

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  • Real-time fluorescence quantification PCR (Polymerase Chain Reaction) kit and method for detecting Y-chromosome micro-deletion
  • Real-time fluorescence quantification PCR (Polymerase Chain Reaction) kit and method for detecting Y-chromosome micro-deletion
  • Real-time fluorescence quantification PCR (Polymerase Chain Reaction) kit and method for detecting Y-chromosome micro-deletion

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Embodiment 1

[0066] 1. Materials

[0067] 1. Instrument: Real-time Fluorescence Quantitative Instrument

[0068] Design of probes and primers: 6 pairs of probes and primers were designed to specifically detect AZF sequences based on the above, and at the same time, 1 pair of probes and primers were designed as quality control to detect male sex-determining genes to detect whether the PCR detection process is normal To carry out, the normal fertile male DNA sample is used as a positive control, the female DNA sample is used as a negative control, and sterilized water is used as a blank control to prevent contamination of the test system.

[0069] 2. Use the following probe primer pairs to detect the deletion of AZFa, AZFb, and AZFc regions.

[0070] SY84 upstream primer 5'-CCTATTTGTTTTAAGGTGCCATTCC-3' (SEQ ID NO: 1)

[0071] SY84 downstream primer 5'-GCTTGCCTGAGCATCCTACAG-3' (SEQ ID NO: 9)

[0072] SY84 probe 5'-VIC-CTCTACCTCCTTCCCCCAGTGCCCA-BHQ-3' (SEQ ID NO: 15)

[0073] SY86 upstream...

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Abstract

Provided is a real-time fluorescence quantitative PCR kit for detecting Y-chromosome microdeletion. The kit comprises a kit body, a PCR reaction solution, and at least one primer probe combination. The primer probe combination is selected from an SY84 primer probe combination for detecting SY84 loci, an SY86 primer probe combination for detecting SY86 loci, an SY127 primer probe combination for detecting SY127 loci, an SY134 primer probe combination for detecting SY134 loci, an SY254 primer probe combination for detecting SY254 loci, and an SY255 primer probe combination for detecting SY255 loci. A method for detecting Y-chromosome microdeletion loci by using real-time fluorescence quantitative PCR in combination with a Taqman probe is further provided.

Description

technical field [0001] The invention relates to a kit and method for detecting Y chromosome microdeletion. Background technique [0002] According to statistics, 15% of couples in the world suffer from infertility (pregnancy), of which more than 50% are caused by the man. In recent years, studies have shown that more than 30% of male infertility is due to spermatogenesis disorders caused by genetic factors, usually manifested as multiple positions in the Azoospermia factor gene family (Azoospermia factor, AZF) on the 11th region of the male Y chromosome long arm (q arm) Point deletion mutations, microdeletions of any loci can lead to spermatogenesis disorders, and male infertility caused by Y chromosome microdeletions cannot be treated. Detection of Y chromosome microdeletions before intracytoplasmic sperm injection (ICSI) assisted fertility treatment can provide guidance for clinical treatment and reduce unnecessary economic waste. At present, relevant societies in Europe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6879C12Q2600/156
Inventor 骆子义邬宇美
Owner 骆子义
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