Nucleic acid aptamer fluorescent probe based alpha-fetoprotein AFP2 kit and detection method

An alpha-fetoprotein and nucleic acid aptamer technology, which is applied in the field of medical testing and determination, can solve the problems of operational technical errors, high testing costs, and many interference factors, and achieves the effects of high repeatability, convenient transportation and strong specificity.

Inactive Publication Date: 2016-06-08
徐大鹏
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  • 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

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  • Nucleic acid aptamer fluorescent probe based alpha-fetoprotein AFP2 kit and detection method

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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 a nucleic acid aptamer fluorescent probe based alpha-fetoprotein AFP2 kit. Simultaneously, the invention also relates to a method for determining concentration of alpha-fetoprotein AFP and determination on composition and ingredients of reagents and belongs to the technical field of medical detection and determination. The kit provided by the invention comprises the main ingredients, i.e., a red blood cell lysis solution, a phosphate buffer solution PBS, an alpha-fetoprotein AFP standard and an alpha-fetoprotein AFP nucleic acid aptamer fluorescent probe; the size of concentration of the alpha-fetoprotein AFP is measured through carrying out blood sample lysis, carrying out mixed egg breeding treatment and carrying out detection by a fluorescence spectrophotometer. The nucleic acid aptamer fluorescent probe based alpha-fetoprotein AFP2 kit and the method have the advantages of simplicity in sample treatment, simplicity and convenience in operation, short detection time, high detection specificity, high sensitivity, high detection result repeatability and the like.

Description

technical field [0001] The invention belongs to the technical field of medical testing and determination, in particular to an alpha-fetoprotein kit and a detection method based on a nucleic acid aptamer fluorescent probe AFP2. 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 mater...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/68
CPCG01N21/6408G01N21/6486G01N33/68
Inventor 徐大鹏
Owner 徐大鹏
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