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Kjeldahl comprehensive syndrome fetal molecule dingnosis reagent box and application

A molecular diagnosis and syndrome technology, which is applied in the field of biomedical technology and can solve problems such as inability to perform prenatal villus or amniotic fluid cell chromosome examinations.

Inactive Publication Date: 2007-05-16
翁亚光
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If a relative of a couple has given birth to a child with Klinefelter syndrome or the couple has given birth to a child with Klinefelter syndrome and wants to have a second child, it is difficult for clinicians to evaluate whether the couple will become pregnant again Syndrome or re-pregnancy Klinefelter syndrome fetal risk, so it is impossible to make a judgment whether it is necessary to collect prenatal villi or amniotic fluid cells for chromosome examination, especially when it is known that the examination may cause miscarriage and the doctor cannot provide the necessary information. When the basis of the inspection, most couples choose to give up the inspection

Method used

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  • Kjeldahl comprehensive syndrome fetal molecule dingnosis reagent box and application

Examples

Experimental program
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Effect test

Embodiment 1

[0081] Example 1: Preparation of Fetal Molecular Diagnosis Kit for Klinefelter Syndrome

[0082] Klinefelter Syndrome Fetal Molecular Diagnosis Kit includes DNA Extraction Solution A, DNA Extraction Solution B, Proteinase K, PCR Reaction Solution, PCR Specific Primer 1, PCR Specific Primer 2, Taq Enzyme, Standard I, Standard II and Positive Control goods, negative control substances, of which:

[0083] DNA extraction solution A is cell lysate, including: EDTA, sodium dodecyl sulfate (SDS), DNase-free RNase lysis buffer.

[0084] DNA extraction solution B is the precipitation solution, usually NH 4 Ac salts.

[0085] The PCR reaction solution contained 2.5 μl of 10×PCR buffer, 2 μl of MgCL, 0.5 μl of dNTP and 17.75 μl of deionized water.

[0086] PCR-specific primer 1 is: sense strand 5′-aag cct ttt cct tta tct tca ca-3′,

[0087] Antisense strand 5′-ctg aga atg ctt ccg ttt gcc-3′,

[0088] PCR specific primer 2 is: sense strand 5′-att ctc aga aag ttt tct gc-3′,

[0089] ...

Embodiment 2

[0117] Example 2: Detection of molecular markers in peripheral blood of parents with Klinefelter syndrome fetal molecular diagnostic kit

[0118] (1) Sample DNA preparation

[0119] 1. Take 1ml of peripheral blood, add sodium citrate for anticoagulation, and then take out 200μl into a 1.5ml sterile EP tube;

[0120] 2. Add 400 μl of DNA extraction solution A to the EP tube, then add 3 μl of proteinase K and mix well, place in a 55°C water bath for 5-10 minutes or longer, mix up and down several times during the process, and adjust the digestion time according to the degradation degree of the sample;

[0121] 3. Add 600 μl chloroform and mix well, not too vigorously, to ensure the integrity of DNA;

[0122] 4. Centrifuge at room temperature for 2 minutes at 10,000 rpm on a desktop high-speed centrifuge;

[0123] 5. Divide into three layers after centrifugation. Genomic DNA is in the upper layer. Take 500 μl of the supernatant and transfer it to a 1.5ml sterile EP tube. The mi...

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Abstract

The invention relates to K. syndrome embryo molecule diagnosing agent box and the application. The agent box includes DNA distill liquid A, DNA distill liquid B, protease K, PCR reaction liquid, PCR specificity 1, PCR specificity 2, Tap enzyme, standard substance and contrast. By testing the molecule label cirumferential blood of the husband and wife before delivery or pregnant by repPCR technology, it supplies reference to determine whether embryo needs to take chromosome examination based on diagnosing K. syndrome.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a Klinefelter syndrome fetal molecular diagnostic kit, which detects molecular markers in the peripheral blood of prenatal or pre-pregnancy couples by using repPCR (representative sampling of multiple repetitive units PCR) technology, for Provides basis for whether the pregnant fetus needs to undergo chromosome examination, and the application of the kit of the present invention in judging whether the pregnant fetus needs to undergo chromosome examination based on the diagnosis of Klinefelter syndrome. technical background [0002] Klinefelt is a congenital disorder caused by an abnormal number of chromosomes. The normal male karyotype is 46, xY, and the female is 46, xx. If there is an increase in x in the karyotype of a man, this disease will be caused. The α-satellite DNA in the entire centromere region of the X chromosome contains a...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 翁亚光李恬施琼涂植光王应雄何俊琳刘学庆陈雪梅杨戎魏莎莉
Owner 翁亚光
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