Alpha fetal protein kit based on aptamer fluorescent probe AFP3 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 many interference factors, and achieve high repeatability, convenient transportation, and strong specificity
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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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