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Reagent kit for detecting chromosome numerical abnormalities resulting in natural abortion

A technology of chromosome number and spontaneous abortion, applied in the field of biochemistry, can solve problems such as unsuitable for large-scale promotion, large probe restrictions, expensive fluorescent probes, etc.

Inactive Publication Date: 2011-03-09
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

All of the above QF-PCR methods can diagnose chromosomal aneuploidy, triploidy, and uniparental diploidy, and identify maternal contamination. However, this method is limited by probes, which are directly affected by the quality of fluorescent probes. As a result, more factors are affected than ordinary PCR, which directly affects the accuracy of the method; in addition, the fluorescent probes used in this method are relatively expensive, which directly limits the clinical application of the method, and the results require special analysis. The software can only interpret the fluorescent signal after processing it, which is not suitable for large-scale promotion

Method used

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  • Reagent kit for detecting chromosome numerical abnormalities resulting in natural abortion
  • Reagent kit for detecting chromosome numerical abnormalities resulting in natural abortion
  • Reagent kit for detecting chromosome numerical abnormalities resulting in natural abortion

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0074] Case 1 Abnormal number of chromosome 2

[0075] 1. Experimental materials:

[0076] (1) The specimens in this case were collected from the villi specimens collected when the pregnant woman underwent uterine curettage after B-ultrasound indicated that the embryo stopped developing (no heartbeat, no fetal buds, etc.) at the 10th week of pregnancy, and the venous blood of both aborted couples was collected at the same time.

[0077] (2) kit of the present invention, its composition is as follows:

[0078] Primers: the primers of the group 1-7 of the present invention were synthesized by Shanghai Handsome Biotechnology Co., Ltd.

[0079] Enzyme: Taq enzyme from Beijing Dingguo Biotechnology Company was used.

[0080] Reagent: 10×buffer (20mM Tris-HCl, 10mM (NH 4 ) 2 SO 4 , 2mM MgCl 2 , 0.1% X-100 and 10 mM KCl, pH 8.8); TAKARA company dNTP (2.5 mM each).

[0081] 2. Experimental method:

[0082] (1) Karyotype analysis:

[0083] Aseptically remove 15-20 mg of vill...

example 2

[0111] Case 2 Abnormal number of chromosome 16

[0112] 1. Experimental materials: The specimens in this case were collected from the villi specimens collected when the pregnant woman underwent uterine curettage after the B-ultrasound indicated that the embryo stopped developing (no heartbeat, no fetal buds, etc.) at the 7th week of pregnancy, and the venous blood of both aborted couples was collected at the same time .

[0113] 2. Experimental method:

[0114] (1) Karyotype analysis:

[0115] Aseptically remove 15-20 mg of villous tissue, roughly cut, finely cut, digested, add medium and inoculate into a culture bottle, 37 ° C, 5% CO 2 Cultivate in the incubator for 5-10 days. Chromosomes in the cells were extracted, and made into glass slides after hypotonicity, pre-fixation, and twice-fixation. After drying, conventional G-banding was performed.

[0116] (2) kit analysis of the present invention:

[0117] a. Use the first group of primers of the kit of the present inve...

example 3

[0144] Example 3 The diagnostic rate statistics of the kit of the present invention

[0145] 1. Experimental materials:

[0146] Villi: 6-12 weeks of gestation, the villi specimens collected when curettage was performed after the B-ultrasound found that the embryo had ceased to be fertile (no heartbeat, no fetal bud, etc.).

[0147] Peripheral blood: Peripheral blood of abortion couples.

[0148] 2. Experimental method:

[0149] (1) Obtaining the theoretical heterozygosity of the selected STR site in the present invention:

[0150] The theoretical heterozygosity of STR loci was searched through the www.gdb.org database.

[0151] (2) Statistics of the actual heterozygous ratio of the selected STR of the present invention:

[0152] The actual heterozygous ratio of STR was calculated by counting the number of heterozygous peaks at each locus after 9 sets of PCR on 120 DNA samples from 40 abortion families (abortion villi DNA and abortion couples' peripheral blood DNA). Actua...

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Abstract

The invention provides a kit for detecting the abnormal number of chromosomes which causes natural abortion. The kit comprises 7 to 9 groups of multiple PCR primers which can carry out molecule detection and source judging on the abnormal number of 10 types of chromosomes (the chromosomes of number 2, number 13, number 15, number 16, number 18, number 20, number 21, number 22, X and Y); and the detection range covers the common abnormal chromosome types during natural abortion and prenatal diagnosis and the kit has the advantages of high delicacy, high accuracy, low detection cost, and the like.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to detection reagents including nucleic acids. Background technique [0002] Spontaneous abortion is a common disease in obstetrics and gynecology, and its incidence accounts for about 10-20% of clinical pregnancies. According to literature reports, direct cytogenetic observation (karyotype analysis) of abortion villi, in <15 weeks of gestation In spontaneous abortion, the incidence of chromosomal abnormalities exceeds 50% (50-70%). In spontaneously aborted embryos, aneuploidy mainly manifests as trisomy and sex chromosome haplotype (autosomal haplotype is extremely rare because of its high lethality), and abnormal karyotypes are mostly trisomy 16, 21 Trisomy, trisomy 18, trisomy 13, trisomy 15 and haplotype X. [0003] At present, the methods for genetic detection of spontaneous abortion embryos reported internationally include: karyotype analysis (G-banding method), FISH (fluoresc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 曾嵘熊丽
Owner SOUTHERN MEDICAL UNIVERSITY
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