Kit for detecting pregnant woman peripheral blood free RNA screening 21-trisomy syndrome based on time-of-flight mass spectrometry

A technology of time-of-flight mass spectrometry and peripheral blood of pregnant women, which is applied in the direction of DNA/RNA fragments, recombinant DNA technology, and microbial measurement/inspection. It can solve the problems of not being included in the scope of medical insurance and high testing costs, and reduce psychological burden and testing costs. Low, easily accessible effects

Active Publication Date: 2020-08-07
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0005]The second-generation high-throughput gene sequencing technology is based on the characteristics of fetal cell-free DNA in the peripheral blood of pregnan

Method used

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  • Kit for detecting pregnant woman peripheral blood free RNA screening 21-trisomy syndrome based on time-of-flight mass spectrometry
  • Kit for detecting pregnant woman peripheral blood free RNA screening 21-trisomy syndrome based on time-of-flight mass spectrometry
  • Kit for detecting pregnant woman peripheral blood free RNA screening 21-trisomy syndrome based on time-of-flight mass spectrometry

Examples

Experimental program
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Effect test

Embodiment 1

[0091] Embodiment 1 kit and its use

[0092] 1. PCR reaction: the purpose of this step is to amplify the DNA fragment where the heterozygous SNP site is located, and the PCR single-hole reaction system is 5ul;

[0093] 1-1. Sample pretreatment (sampling and sample preparation): The peripheral blood samples of pregnant women are extracted according to the Qiagen RNA extraction and reverse transcription kit instructions. Store cDNA samples at -20°C when in use, and store at 4°C when in use;

[0094] 1-2. Dilute the PCR forward and reverse primers (dry powder) with TE buffer, configure 1uM (each primer) PCR primer mixture, store at -20°C, and store all other reagents at -20°C;

[0095] 1-3. The PCR mixture reaction system (total reaction system 5ul) is specifically:

[0096]

[0097] 1-4. Use a pipette to add 5uL of the prepared PCR mixture to each well of the PCR96-well plate, and then mix well; seal the plate with a sealing film, vortex twice and centrifuge (4000rpm, centrif...

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Abstract

The invention discloses a kit for detecting pregnant woman peripheral blood free RNA screening 21-trisomy syndrome based on time-of-flight mass spectrometry. The kit comprises a PCR reaction system and a single base extension reaction system. The kit provided by the invention is combined with time mass spectrometry detection, and has the following advantages: 1, the detection accuracy is high; 2,the detection flux is high; 3, the detection sensitivity is high; 4, the detection sites are flexible and variable; 5, the detection process is simple, and the operation time is short; 6, the detection cost is low; 7, prenatal non-invasive detection is performed, damage to the pregnant woman is extremely small, psychological burdens of the pregnant woman are reduced, and a detection sample is peripheral blood of the pregnant woman and is easy to obtain; and 8, the whole detection operation is easy to repeat, mass spectrum chip sample application is fully automatic, the detection result is automatically analyzed through software, and errors caused by improper manual operation can be reduced.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a kit for screening free RNA in peripheral blood of pregnant women based on time-of-flight mass spectrometry for screening 21-trisomy syndrome. Background technique [0002] Chromosomal aneuploidy is a common chromosomal disease, including 21-trisomy syndrome, 18-trisomy syndrome, 13-trisomy syndrome, etc. Among them, trisomy 21 is the most common type, with an annual incidence rate of 1 / 800-1 / 600, accounting for about 70%-80% of trisomy chromosomal diseases in children, and its incidence increases with the increase of mother's age. Children are mainly manifested as mental retardation, backward growth and development, some combined deformities, and dysplasia of multiple organs, which bring a heavy burden to the family and society. At present, non-invasive prenatal screening measures are mainly used to determine high-risk groups clinically; commonly used non-invasive...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6872C12Q1/686C12N15/11
CPCC12Q1/6883C12Q1/6872C12Q1/686C12Q2600/156
Inventor 刘派董文飞王昇赵也明常智敏李力梅茜
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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