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Alpha fetoprotein kit based on aptamer fluorescent probe AFP4 and detection method thereof

A technology of alpha-fetoprotein and nucleic acid aptamer, which is applied in the field of medical testing and determination, can solve problems such as operating technical errors, poor repeatability, and high testing costs, and achieve the effects of high repeatability, convenient transportation, and high sensitivity

Inactive Publication Date: 2016-05-25
徐大鹏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The purpose of the present invention is to provide a nucleic acid aptamer fluorescent probe based alpha-fetoprotein AFP kit and Its detection method

Method used

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  • Alpha fetoprotein kit based on aptamer fluorescent probe AFP4 and detection method thereof

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Experimental program
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Effect test

Embodiment 1

[0033] After dissolving and preparing the required reagent components according to Example 1 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:

[0034] (1) Blood sample lysing: Mix heparin anticoagulated whole blood and erythrocyte lysate at a volume ratio of 1:0.5, let stand for 30 minutes, then centrifuge at a medium speed for 7 minutes, and collect the supernatant;

[0035] (2) Mixed egg incubation: Take 20μl of the supernatant obtained in step 1) and mix with 45ul of the AFP nucleic acid aptamer fluorescent probe reagent obtained by dissolving the AFP nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer, and mix at room temperature Eggs were incubated for 5 minutes to obtain the test solution;

[0036] (3) Fluorescence detection: 50ul of the test solution obtained in the detection step 2) of ...

Embodiment 2

[0040] After dissolving and preparing the required reagent components according to Example 2 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:

[0041] (1) Blood sample lysis: Mix peripheral blood and red blood cell lysate at a volume ratio of 1:2.5, let stand for 5 minutes, then centrifuge at a medium speed for 6 minutes, and collect the supernatant;

[0042] (2) Mixed egg incubation: Take 50μl of the supernatant obtained in step 1) and 30ul of

[0043] The alpha-fetoprotein nucleic acid aptamer fluorescent probe reagent obtained by dissolving the alpha-fetoprotein nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer was mixed, and the eggs were incubated at room temperature for 6 minutes to obtain the test solution;

[0044] (3) Fluorescence detection: 50ul of the test solution obtained in the ...

Embodiment 3

[0047] After dissolving and preparing the required reagent components according to Example 3 in Table 1, divide into bottles and freeze-dry to make a dry powder reagent; before use, add ultrapure water and use after reconstitution. Three parallels were set for each sample, and the detection steps were as follows:

[0048] (1) Blood sample lysis: Mix peripheral blood and red blood cell lysate at a volume ratio of 1:3.5, let stand for 10 minutes, then centrifuge at a medium speed for 5 minutes, and collect the supernatant;

[0049] (2) Mixed egg incubation: Take 75μl of the supernatant obtained in step 1) and 45ul of

[0050] The alpha-fetoprotein nucleic acid aptamer fluorescent probe reagent obtained by dissolving the alpha-fetoprotein nucleic acid aptamer fluorescent probe in 0.2M phosphate buffer was mixed, and the eggs were incubated at room temperature for 7 minutes to obtain the test solution;

[0051] (3) Fluorescence detection: 50ul of the test solution obtained in the...

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Abstract

The invention relates to an alpha fetoprotein kit based on an aptamer fluorescent probe AFP4, a method for determining the alpha fetoprotein AFP concentration and compositions and ingredients of a determining reagent and belongs to the technical field of medical examination determination. The kit provided by the invention mainly comprises an erythrocyte lysis solution, a phosphate buffer PBS, an alpha fetoprotein AFP standard substance and an alpha fetoprotein AFP aptamer fluorescent probe. The concentration of the alpha fetoprotein AFP can be measured through blood sample splitting decomposition and mixing spawn raising treatment as well as fluorescence spectrophotometer detection. The alpha fetoprotein kit provided by the invention has the advantages of simple sample treatment, convenience in operation, short detection time, strong detection specificity, high sensitivity, high repeatability of detection results, and the like.

Description

technical field [0001] The invention belongs to the technical field of medical examination and determination, in particular to an alpha-fetoprotein kit and a detection method based on a nucleic acid aptamer fluorescent probe AFP4. Background technique [0002] Alpha-fetoprotein (AFP) is a glycoprotein. Under normal circumstances, this protein mainly comes from the liver cells of the embryo. About two weeks after the fetus is born, AFP disappears from the blood. Therefore, the content of AFP in normal human serum Less than 20 micrograms / liter. Alpha-fetoprotein in maternal amniotic fluid or maternal plasma AFP can be used for fetal prenatal monitoring. For example, in neural tube defects, spina bifida, anencephaly, etc., AFP can enter the amniotic fluid through the open neural tube, resulting in a significant increase in its content in amniotic fluid. Fetal death in the uterus, congenital defects such as teratoma may also increase AFP in amniotic fluid. AFP can enter the m...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/68
CPCG01N21/6408G01N21/6486G01N33/68
Inventor 徐大鹏
Owner 徐大鹏
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