Detection kit for human Y-chromosomal microdeletion and application of detection kit

Abstract

The invention provides a multi-PCR amplification system and detection kit for human Y-chromosomal microdeletion. The system comprises amplification primers comprising following loci with the corresponding sequences from SEQ ID No.1 to SEQ ID No.40, wherein the loci comprise 15 Y-chromosomal microdeletion loci (4 in the AZFa region, 4 in the AZFb region, 5 in the AZFc region and 2 in AZFd region),3 sample molecular tag loci (two X-STR loci and one autosomal locus) and 2 gender quality control loci (SRY, ZFX / Y). The 15 Y-chromosomal microdeletion loci use sequence tag loci (STS) specific primers, the 3 sample molecular tag loci use length polymorphism primers, and the 2 gender quality control loci use STS specific primers (EAA / EMQN Y-chromosomal microdeletion related aboratory optimal handbook recommendation detection loci, 2013). The primers are compatible with one another and can react in the same PCR amplification system. In addition, the invention further provides the detection kitcontaining an amplification primer composition container, a PCR reaction mother liquor container and a PCR auxiliary liquid container and application of the detection kit in human Y-chromosomal microdeletion non-diagnostic detection. After DNA extraction, target fragments of 20 genetic marker loci are amplified through a PCR reaction, the cost, labor and time can be remarkably saved, and the workefficiency is improved.

Description

Technical field [0001] The invention belongs to the technical field of human nucleic acid in vitro detection, and specifically relates to a multiple PCR amplification system and a detection kit for human Y chromosome microdeletion. Background technique [0002] According to statistics from the World Health Organization (WHO), about 10-15% of couples worldwide suffer from infertility, and male infertility accounts for half. Among male infertility, more than half are of unknown cause. Among these patients, about 15% have azoospermia and oligospermia. Except for vas deferens obstruction, endocrine abnormalities, sexual dysfunction, congenital diseases, infections and other causes, the cause of a considerable number of patients has been unclear. Since Tiepolo and Zuffardi discovered that there is a distal Yq11 deletion in azoospermous patients in 1976, more and more reports support the presence of spermatogenesis regulatory genes at the distal end of Yq11. Their deletion or mutation...

AI Technical Summary

Problems solved by technology

In view of the different operating platforms or conditions of different laboratories, there are three main detection methods: PCR-gel electrophoresis method, which is a traditional molecular detection method, that is, the detection of target bands after end-point PCR and agarose gel electrophoresis. The judgment of results given by naked eye observation is generally restricted by factors such as PCR product concentration and agarose gel concentration, and is easily affected by false positives and misjudgments caused by specific electrophoretic bands. Overall, the overall PCR amplification conditions and result judgments have a greater impact. High requirements, difficult to promote clinical application; PCR fluorescent probe method, which uses different fluorescently labeled probe combinations for rapid amplification and real-time quantification of target fragments, is currently a commonly used technical method in the field of rapid molecular diagnosis, usually Amplify the target fragment by multi-tube reaction, collect the fluorescent signal of the amplification curve and analyze the CT value. Rapid screening of diseases by multi-site detection; MLPA method, the principle of which is based on molecular hybridization and multiplex PCR combined with capillary electrophoresis detection method, is a relatively certain molecular rapid diagnosis method at present, but due to expensive reagent consumables and long operation time It is long and requires high experimental operation, which is relatively difficult in actual clinical promotion

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