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Plasma room-temperature transportation method for detecting fetal free DNA

A technology for separating plasma at normal temperature, which is applied in the field of plasma free nucleic acid sequencing, which can solve problems such as mechanical collision or shaking, increased testing costs, cold chain or dry ice transportation temperature out of control, etc., to avoid overtime and improve inconvenient effects

Inactive Publication Date: 2018-05-01
CAPITALBIO GENOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, anticoagulant blood collection tubes or free nucleic acid stable tubes are usually selected according to the actual situation when collecting peripheral blood from pregnant women. Among them, EDTA anticoagulant blood collection tubes need to be stored at 4°C after collection, and must be delivered to the laboratory for plasma separation within 8 hours; if Use free nucleic acid stabilization tubes that are more than ten to dozens of times more expensive than EDTA anticoagulant blood collection tubes, such as Streck cfDNA BCT blood collection tubes, which can be stored and transported at room temperature, but still need to be delivered within 72 to 96 hours For plasma separation in the laboratory, due to the instability of logistics transportation time and mechanical collision or shaking during transportation, peripheral blood samples often have hemolysis caused by transportation overtime or mechanical force, and peripheral blood needs to be re-collected
[0005] For a long time, if the plasma samples that have completed plasma separation need to be transported to the testing point, they must rely on high-cost cold chain logistics transportation or dry ice insulation transportation. Chain or dry ice transportation temperature is out of control, and the price of dry ice is expensive, which increases the detection cost

Method used

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  • Plasma room-temperature transportation method for detecting fetal free DNA
  • Plasma room-temperature transportation method for detecting fetal free DNA
  • Plasma room-temperature transportation method for detecting fetal free DNA

Examples

Experimental program
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Effect test

specific Embodiment 1

[0035] 1. Normal temperature transport and treatment of blood plasma

[0036] Peripheral blood was collected from 3 pregnant women and placed in EDTA anticoagulant tubes, and placed at 4°C for 8 hours to separate plasma by centrifugation. The centrifugation steps were: whole blood was centrifuged at 1600g for 10 minutes, and the supernatant was carefully sucked to avoid white blood cells. The supernatant was placed in In a centrifuge tube, centrifuge at 16000g for 10min, the supernatant is the separated plasma, carefully draw the supernatant and put it into a sterile nuclease-free EP tube, and seal it.

[0037]The peripheral blood was collected from 3 pregnant women and placed in the Cell Free DNA preservation tube of Wanji Bio (hereinafter referred to as Wanji preservation tube), and the plasma was separated by centrifugation within 72 hours. To avoid absorbing white blood cells, put the supernatant in a centrifuge tube and centrifuge at 16,000g for 10 minutes. The supernatan...

specific Embodiment 2

[0084] 1. Normal temperature transport and treatment of blood plasma

[0085]Peripheral blood was collected from 3 pregnant women and placed in EDTA anticoagulant tubes, and placed at 4°C for 8 hours to separate plasma by centrifugation. The centrifugation steps were: whole blood was centrifuged at 1600g for 10 minutes, and the supernatant was carefully sucked to avoid white blood cells. The supernatant was placed in In a centrifuge tube, centrifuge at 16000g for 10min, the supernatant is the separated plasma, carefully draw the supernatant and put it into a sterile nuclease-free EP tube, and seal it.

[0086] The peripheral blood collected from 3 pregnant women was placed in the Cell Free DNA preservation tube of Wanji Bio (hereinafter referred to as the Wanji Bio preservation tube), and the plasma was separated by centrifugation within 72 hours. Supernatant, avoid absorbing white blood cells, put the supernatant in a centrifuge tube, centrifuge at 16000g for 10min, the super...

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Abstract

The invention discloses a processing method of plasma room-temperature transportation, wherein plasma is separated from maternal peripheral blood, put into a sterile nuclease-free device, sealed, andthen can be transported at room temperature. After transportation at the room temperature, the original free DNA concentration, free DNA fragment distribution, fetal free DNA percentage concentration,sequencing fragment length distribution, and chromosome Z detection value of the plasma are all equivalent to those of the plasma stored at ultra-low temperature (minus 70 DEG C), and the plasma transported at the room temperature is suitable for detecting the fetal free DNA and noninvasive prenatal testing (NIPI) products. The plasma room-temperature transportation method for detecting fetal free DNA, disclosed by the invention, does not need a special device and does not need to add a protective agent additionally, sample transportation of the plasma can be expanded to normal-temperature transportation, the method improves the problems of inconvenience and high transportation cost of plasma cold chain transportation, and can avoid the embarrassment of overtime transportation of a wholeblood sample and hemolysis due to a transport mechanical force, and a novel, effective and convenient alternative method is provided for the existing maternal peripheral blood processing mode and plasma transportation mode.

Description

technical field [0001] The invention belongs to the field of plasma free nucleic acid sequencing, and more specifically relates to a normal-temperature transport processing method for plasma and its application. Background technique [0002] In 1997, Professor Lu Yuk-ming of the Chinese University of Hong Kong discovered the presence of fetal cell-free DNA in the peripheral blood of pregnant women, and since then created a new era of noninvasive prenatal testing (Noninvasive Prenatal Testing, NIPT). At present, based on maternal peripheral blood samples and high-throughput sequencing technology, non-invasive fetal chromosomal aneuploidy screening, non-invasive fetal chromosomal microdeletion and microduplication screening have been applied to clinical services, and non-invasive fetal single-gene disease screening technology is being developed and developed. Sample accumulation verification phase. Compared with traditional invasive screening / diagnostic techniques, such as am...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/02A01N1/0263
Inventor 糜庆丰罗东红谢婵芳陈样宜黄铨飞彭春方饶兴蔷刘丽菲
Owner CAPITALBIO GENOMICS
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