Primer group, method and kit for detecting triploids of fetus

A technology of triploid and primer set, which is applied in the direction of biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of high requirements for PCR reaction conditions, large number of primers, high cost, etc. Quality, process and data analysis are simple and cost-effective

Active Publication Date: 2015-05-06
PEKING JABREHOO MED TECH CO LTD
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  • Claims
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AI Technical Summary

Problems solved by technology

However, due to the large number of primers, the cost is high and the conditions for PCR reactions are high
[0006] T...

Method used

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  • Primer group, method and kit for detecting triploids of fetus
  • Primer group, method and kit for detecting triploids of fetus
  • Primer group, method and kit for detecting triploids of fetus

Examples

Experimental program
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Effect test

Embodiment 1

[0050] (1) Separation and extraction of cfDNA:

[0051] Collect 10ml of venous blood from pregnant women (12-24 weeks of pregnancy) into vacuum blood collection tubes (EDTA anticoagulant), centrifuge at 1600g for 10 minutes, and centrifuge at 16000g for 10 minutes to obtain about 4ml of plasma, and use a free DNA extraction kit to separate free DNA in the plasma , to get 15ng cfDNA.

[0052] (2) Primer design:

[0053] For human chromosomes 13, 18, and 21, 300 pairs of sequence-specific primers were designed respectively. The sequence-specific primer has the following characteristics: (1) Each site of the primer is composed of two consecutively arranged specific position oligos dependent on peripheral blood free DNA (cfDNA) through an intermediate oligonucleotide bridge Form a PCR template; (2) the primers have a unique sequence on the target chromosome; (3) the position-specific oligonucleotides have the same annealing temperature; The universal primers have the lowest seq...

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Abstract

The invention relates to a primer group, a method and a kit for detecting the triploids of a fetus by virtue of plasma free DNS in maternal peripheral blood, wherein the DNA is selected from human No. 21, No. 18 and No. 13 chromosomes, 300 pairs of sequence specific primers are designed with regard to the No. 21, No. 18 and No. 13 chromosomes respectively, each pair of the sequence specific primers comprises a forward primer, a reverse primer and a middle primer, and the gene sequence numbers of the primers are SEQ ID NO: 1-2700. The primer group, the method and the kit disclosed by the invention have the following beneficial effects: the triploids of the fetus are detected by the method disclosed by the invention, the detection is non-invasive detection without a threat on the health of the fetus, diagnosis and detection are high in accuracy and low in expense, flow and data analysis are simple and convenient, and PCR reaction condition requirements are not rigorous; the primer group, the method and the kit are wide in applicability, can be used for promoting the popularization of non-invasive screening for a trisomic syndrome, and are conductive to improving the population quality of China.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a product, method and kit for detecting fetal triploids by using plasma free DNA in maternal peripheral blood, that is, using PCR technology and sequencing technology to detect free DNA from maternal peripheral blood Test to determine whether the fetus is triploid. Background technique [0002] Aneuploidy detection of fetal chromosomes is one of the important prenatal examinations [1,2] . Prenatal care currently includes screening and diagnosis. Screening includes analysis of serum markers, fetal ultrasonography (such as ultrasound measurement of fetal nuchal translucency thickness), but sensitivity and specificity need to be improved [1] . Diagnostic testing includes invasive chorionic villus sampling (CVS) or amniocentesis. Current screening methods are not yet accurate enough to be used for censuses. Diagnostic tests, while highly accurate, are invasive and risk misc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6883C12Q2600/158C12Q2563/143
Inventor 冯涛
Owner PEKING JABREHOO MED TECH CO LTD
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