Primer composition for detecting Y chromosome microdeletion and sex chromosome number and application

Abstract

The invention relates to the field of molecular biology diagnosis, in particular to a primer composition for detecting Y chromosome microdeletion and the sex chromosome number and an application. A Ychromosome AZF region sequence marker site and a sex chromosome number detection site can be simultaneously amplified by utilizing special primer design in a tube of PCR reaction solution; a male sexdetermination gene (SRY) is used as a sex abnormality control and a human X / Y linked zinc finger protein gene (ZFX / Y) is used as an internal control; PCR products are analyzed through capillary electrophoresis; and whether an AZF region of Y chromosome is deleted or not and whether the sex chromosome number is abnormal or not are interpreted by comparing the PCR products with different fragment sizes. A result can be obtained within 3 hours from a DNA sample, the Y chromosome microdeletion and the Klinefelter syndrome can be detected at the same time only through one PCR experiment, the detection technical process is simple, and standardization is easy to achieve.

Description

technical field [0001] The invention relates to the field of molecular biology diagnosis, and relates to a primer composition and application for detecting Y chromosome microdeletion and sex chromosome number. Background technique [0002] According to WHO statistics, 10-15% of couples in the world suffer from infertility, of which male factors account for 50%, and genetic factors account for 30% of male infertility. Among patients, Klinefelter syndrome is the most common genetic factor, followed by Y chromosome microdeletion. [0003] Klinefelter syndrome (KS) is the most common sex chromosome abnormality disease, also known as congenital testicular hypoplasia syndrome, which is non-disjunction of sex chromosomes during meiosis, resulting in two or more X chromosomes in males, About 90% of patients have a karyotype of 47XXY, and 10% have 47,XXY / 46,XY mosaic, 48XXXY, 48XXYY, 49XXXXY and structural sex chromosome abnormalities. The clinical phenotypes of patients are diverse...

AI Technical Summary

Problems solved by technology

MLPA combines DNA probe hybridization and PCR technology, can detect 40 or more sites in a single reaction, and can detect point mutations; The operator has high requirements, and the whole experiment process takes about two days, which is more time-consuming than other detection methods

[0009] Fluorescence in situ hybridization (Fluorescence in situ hybridization) is to hybridize the nucleic acid probe directly or indirectly labeled with fluorescein with the nucleic acid sequence in the sample to be tested according to the principle of complementary base pairing, and directly observe it under a fluorescence microscope after washing Compared with traditional radiolabeled in situ hybridization, it has the advantages of fast operation, strong detection signal, high specificity and reproducible staining, but compared with multiplex PCR, real-time quantitative fluorescent PCR, chip method, etc., the detection steps Complicated, long cycle, low sensitivity and specificity, limited by microscope resolution, high requirements for operators to interpret

[0010] Routine diagnostic methods for Klinefelter syndrome include routine serological examinations and semen examinations. The diagnosis requires karyotype analysis. Karyotype analysis is performed by staining the chromosomes after being treated with protease. Sorting and numbering of chromosomes based on features such as point positions, and diagnosing whether chromosomes are abnormal through the structure or number of chromosomes. Chromosome karyotype analysis requires a large amount of samples, and amniotic fluid cells need to be cultured in vitro. Cell growth speeds vary, and chromosome staining is performed. Sorting and sorting is time-consuming, and it takes an average of one to two weeks to get the results. There are certain requirements for amniotic fluid extraction technology and chromosome identification, and the cost of time and manpower is quite high.

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