Method and kit for analyzing ploidy of unicellular sex chromosomes quickly

Abstract

The invention relates to a method and kit for analyzing the ploidy of unicellular sex chromosomes quickly. According to the method, primers, as shown in SEQ ID NO.1-6, for X chromosome, Y chromosome and chromosome 21 respectively are adopted for preamplification of trace DNA from a single cell firstly, then nested primers as shown in SEQ ID NO.7-12 are adopted for real-time fluorescent quantitative PCR to obtain the DNA content values K and L of X chromosome and Y chromosome relative to the internal reference, a male standard substance, a female standard substance and a negative control group are arranged at the same time, and finally the values of K / L, K / K [female standard], L / L[female standard], K / K [male standard] and / or L / L[male standard] are calculated and the ploidy of the sex chromosomes of a sample to be tested is obtained. The method has the advantages of being high in accuracy, short in time and low in cost when used for analyzing the ploidy of the unicellular sex chromosomes and is helpful for clinical genetic diagnosis.

Description

【Technical field】 [0001] The invention relates to the field of biotechnology, in particular to a method and kit for rapidly analyzing unicellular sex chromosome ploidy. 【Background technique】 [0002] In vitro fertilization-embryo transfer, referred to as test-tube baby technology, has the opportunity to obtain the cells of an embryo before embryo transfer, thus providing conditions for pre-implantation genetics and sex screening of embryos. However, in our country and many European countries, the law does not allow purely gender screening. However, in some countries, such as the United States, sex screening before embryo transfer is permitted by law. However, sex screening to exclude sex-linked genetic diseases is permitted by law. For example, 47,XXY Klinefelter syndrome with doubled X chromosome has an incidence rate of 1 / 850 male newborns; female 45,XO, Turner syndrome with doubled X chromosome deletion has an incidence rate of 1 / 850 among female babies 1 / 2500-5000; I...

AI Technical Summary

Problems solved by technology

However, such a chip is expensive and takes a long time (1-2 days), and the hospital itself cannot operate it. It has to be sent to a company for testing, which increases the chance of sample contamination. The cost of testing an embryo blastomere is about 6,000 yuan. , not suitable for most IVF patients

[0004] In general, the problems with existing screening methods are high cost, long time, and many turnover laboratories

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