Preservation solution for peripheral blood samples of pregnant women and preservation method for peripheral blood samples of pregnant women

A technology of peripheral blood and peripheral blood of pregnant women, applied in the field of blood sample preservation, can solve the problems of inability to meet the needs of non-invasive prenatal diagnostic tests, NRBC preservation damage, etc., and achieve the effect of ensuring accuracy and delaying apoptosis

Active Publication Date: 2021-01-05
BGI SHENZHEN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the NRBC content in the peripheral blood of pregnant women is very rare, when the peripheral blood samples of pregnant women are preserved with Alfred's solution, because the cells are still active and the metabolism of the blood is still going on, it will cause irreversible preservation of the originally rare NRBC. Injury, severe or even unable to meet the needs of subsequent non-invasive prenatal diagnostic tests

Method used

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  • Preservation solution for peripheral blood samples of pregnant women and preservation method for peripheral blood samples of pregnant women
  • Preservation solution for peripheral blood samples of pregnant women and preservation method for peripheral blood samples of pregnant women
  • Preservation solution for peripheral blood samples of pregnant women and preservation method for peripheral blood samples of pregnant women

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] In this example, a comparative test was carried out on the components added to fresh blood samples. Take a tube of fresh pregnant women's peripheral blood for density gradient centrifugation and observe the centrifugation effect and cell status as a reference. Then set, test group a: fresh peripheral blood of pregnant women, without any addition. Experimental group b: fresh peripheral blood of pregnant women was added with Alfred's solution, and the volume ratio of pregnant women's peripheral blood to Alfred's solution was: peripheral blood of pregnant women: Alfred's solution = 1:15. Test group c: fresh peripheral blood of pregnant women, added with Caspase inhibitors, the volume ratio of peripheral blood of pregnant women to Caspase inhibitors is: peripheral blood of pregnant women: Caspase inhibitors = 1:0.1. Experimental group d: fresh peripheral blood of pregnant women was added with a preservation solution composed of Alfred's solution and Caspase inhibitor. The ...

Embodiment 2

[0045] Two tubes of fresh pregnant women’s peripheral blood were drawn from the hospital, one of which was mixed with Caspase inhibitors and then transported to the laboratory at low temperature. The volume ratio of pregnant women’s peripheral blood to Caspase inhibitors was: Peripheral blood of pregnant women: Caspase inhibitors=1 -0.1. After being transported to the laboratory, the peripheral blood of the pregnant woman was added to the Alfred's solution according to the volume ratio of the pregnant woman's peripheral blood: Alfred's solution = 1:3, and stored in a refrigerator at 4°C. After 1 day, 2 days, 3 days, 5 days, and 7 days of storage, observe the morphological integrity of the cells in the blood samples and the cell viability detected by FDA+Hochest fluorescence, and perform gradient centrifugation to observe the centrifugation effect.

[0046] Another tube of pregnant women’s peripheral blood was directly stored in a 4°C refrigerator for 3 days. This sample was co...

Embodiment 3

[0051] On the basis of Example 2, in this example, Alfred's solution and Caspase inhibitors were also made into a preservation solution for testing. Specifically, 1. Preservation solution prepared in the laboratory: 100ul of caspase apoptosis inhibitor was added to 15ml of Alfred's solution, the final concentration was 0.67%, mixed well, stored at -80°C in a 50ml centrifuge tube, taken out and melted before sampling room temperature. 2. Carry a 50ml preservation solution tube to the hospital at room temperature, and take peripheral blood samples from pregnant women. The pregnancy week is about 10-20 weeks. Use a 5ml heparin sodium tube, shake well and immediately transfer to a 50ml preservation solution tube. 3. The sample is placed in a low-temperature sample delivery box and quickly transported from the hospital to the laboratory, and stored in a refrigerator at 4°C. Other settings are the same as in Embodiment 2.

[0052] The results showed that the morphological integrit...

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Abstract

Disclosed are a preserving solution for a maternal peripheral blood sample and method for preserving the maternal peripheral blood sample. The preserving solution of the present invention consists of an Alsever solution and an apoptosis inhibitor at a volume ratio of 5-15:0.03-0.1. The preserving solution of the present invention utilizes the Alsever solution and the apoptosis inhibitor for the preservation of maternal peripheral blood, thus effectively delaying apoptosis in the maternal peripheral blood and maintaining various indexes of a cell to keep the cell fresh as far as possible; maintaining cell morphology, providing nutrients for the cell and maintaining various physicochemical properties of the cell; providing a basis for the gradient centrifugation and flow sorting of subsequent nucleated red blood cells and other rare cells, and ensuring preservation of a large amount of fresh nucleated red blood cells and accurate non-invasive antenatal diagnosis.

Description

technical field [0001] The present application relates to the field of blood sample preservation, in particular to a preservation solution for preservation of peripheral blood samples of pregnant women and a method for preservation of peripheral blood samples of pregnant women. Background technique [0002] Several methods are currently available for prenatal diagnosis of genetic disorders. According to whether the sampling route or diagnostic method constitutes an impact on the fetus, it can be divided into invasive prenatal diagnosis and non-invasive prenatal diagnosis. Invasive prenatal diagnosis includes amniocentesis, chorionic villous cell aspiration, and fetal umbilical cord blood puncture; although the diagnosis accuracy is high, it can lead to complications such as miscarriage and can only be performed in high-risk pregnant women. Non-invasive prenatal diagnosis is mainly for prenatal diagnosis of fetal cells entering the mother's blood. The advantage of this diagn...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02C12N5/078
CPCA01N1/02A61K35/18
Inventor 邓乐潘健昌刘萍
Owner BGI SHENZHEN CO LTD
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