Y chromosome microdeletion detection kit and method

Abstract

The invention relates to the field of chromosome deletion detection, in particular to a Y chromosome microdeletion detection kit and a method, wherein the kit comprises a kit body, a PCR reaction solution I, a PCR reaction solution II, a positive control, a negative control and a blank control. According to the kit, gene deletion of sY84 and sY86 sites of an AZFa subarea of Y chromosome, sY127 and sY134 sites of an AZFb subarea of the Y chromosome and sY254 and sY255 sites of an AZFc subarea of the Y chromosome is detected by virtue of a Taqman fluorescent probe PCR method. The kit, by virtue of specific primer probes which are designed in accordance with the detection sites, can achieve efficient and stable multi-PCR detection on the detection sites, and the various detection site probes are marked by virtue of different fluorescent genes so as to indicate whether the circumstance of deletion occurs in a target segment or not, so that six AZF deleted sites are simultaneously detected in reaction solutions in two tubes and SRY and ZFY internal control genes are simultaneously detected in the reaction solution in each tube. With the implementation of the kit disclosed by the invention, six sites of male Y chromosome microdeletion can be detected; and the kit has the advantages of being accurate, rapid, high in throughput, free from pollution and the like.

Description

technical field [0001] The invention relates to the field of chromosome deletion detection, in particular to a Y chromosome microdeletion detection kit and method. Background technique [0002] It is estimated that infertility afflicts more than 10% of couples of childbearing age in the world, of which male factors account for half. There are many causes of male infertility, including infection, malformation of reproductive organs, abnormal immune function, varicocele, erectile dysfunction, drug damage and chromosomal abnormalities. Male infertility is often accompanied by a decrease in sperm count, and domestic scholars have done a lot of related research on it. Azoospermia factor (AZF) microdeletion on the long arm of the Y chromosome is one of the main genetic factors leading to male spermatogenesis disorders. Studies have shown that the incidence of AZF microdeletion in male infertility patients in Asia is between 2% and 19.4%, which is related to the inclusion criteri...

AI Technical Summary

Problems solved by technology

[0003] The techniques currently used to detect microdeletions on the Y chromosome are mainly the specific amplification of the sequence tag site in the AZF region of the Y chromosome by the PCR method, and then the detection of the PCR product by electrophoresis or Southern blot hybridization. Although the method is mature, there are also defects. The electrophoresis detection method has low accuracy and low sensitivity, and the Southern blot hybridization method is time-consuming, labor-intensive, expensive and cumbersome to operate.

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