Test strip for detecting alpha fetoprotein in whole blood quickly and manufacturing method of test strip

Abstract

The invention discloses a test strip for detecting alpha fetoprotein in whole blood quickly and a manufacturing method of the test strip and belongs to the field of biological test product manufacturing. The test strip is based on a colloidal gold immune chromatography technology and has the advantages of being fast in detection, high in sensitivity, high in specificity, good in stability and easy and convenient to operate, no professional personnel is needed in the detection process, dependence on large instruments is not needed, the content of alpha fetoprotein in the blood can be detected quickly at any time just by 80 microliters of blood, the detection range of the test strip is 5ng-200 ng / ml, the detection time is shorter than 5 minutes, and the test strip can be widely applied to detection of the content of alpha fetoproteinin in blood in physical examination (tumor screening) and family self-examination.

Description

technical field

[0001] The invention belongs to the field of biological test product manufacture, and in particular relates to a test strip for rapidly detecting alpha-fetoprotein in whole blood and a preparation method thereof. Background technique

[0002] The rapid and accurate detection of liver cancer specific markers is of great significance for the early diagnosis and treatment of primary liver cancer. Under normal circumstances, alpha-fetoprotein (AFP) mainly comes from the liver cells of the embryo. About two weeks after the fetus is born, alpha-fetoprotein disappears from the blood. The serum alpha-fetoprotein of normal people is still less than 20ng / ml. During the period of liver cell regeneration, some embryonic carcinomas, germ cell tumors, digestive tract cancers, and AFP in the blood of pregnant women also increase to varying degrees. But the most noteworthy is that in patients with liver cancer, the concentration of AFP in the blood can be greatly increased,...

Examples

preparation example Construction

[0036] The preparation method of the test strip for rapid detection of alpha-fetoprotein in whole blood disclosed by the present invention comprises the following steps:

[0037] 1) Colloidal gold particles with uniform shape and size were prepared by the citrate reduction method of chloroauric acid, and the diameter of the particles was 20 nm.

[0038] 2) Prepare the colloidal gold immunoprobe, adjust its pH to 8.5-9.0, and modify the AFP monoclonal antibody 2 according to the principle that high-concentration salt solution can cause coagulation of unstable colloidal gold.

[0039] When modifying, mix 0.6-1.5ml of colloidal gold and 10-15μg of AFP monoclonal antibody 2 evenly and let stand at 4°C for 25-35 minutes. Add BSA for blocking for 35-45 minutes, keeping its final concentration at 0.5%-1.5%. Centrifuge at 10000-14000rpm under centrifugation conditions, and use PBS buffer containing 0.5%-1.5% BSA, 0.03%-0.1% sodium azide, 0.5%-1.5% sucrose, and 0.5%-1.5% Tween-20 for ...

Embodiment 1

[0044] 1) Preparation of 20nm colloidal gold: Take 100 mL of 0.01% (v / v) chloroauric acid solution, put it into a cleaned and dried Erlenmeyer flask, place it on a heating magnetic stirrer and heat it to boiling. Maintain the heating state and accurately and quickly add 2.0 ml of freshly prepared 1% (v / v) trisodium citrate solution in one go under vigorous stirring. It can be observed that the color of the solution changes from yellow→black→purple→dark blue→wine red, and continue stirring for 15 minutes until the solution turns transparent wine red. After the reaction solution was cooled to room temperature, it was stored at 4°C.

[0045] 2) Preparation of immune probe: first use 0.1M k 2 CO 3 The pH of the colloidal gold was adjusted to 8.7, and 13 μg of AFP monoclonal antibody 2 was added to 1 ml of the colloidal gold solution and left to stand at 4° C. for 25 minutes. 10% (v / v) BSA was added for blocking for 40 minutes, keeping its final concentration at 1%. Make the im...

technical field

[0001] The invention belongs to the field of biological test product manufacture, and in particular relates to a test strip for rapidly detecting alpha-fetoprotein in whole blood and a preparation method thereof. Background technique

[0002] The rapid and accurate detection of liver cancer specific markers is of great significance for the early diagnosis and treatment of primary liver cancer. Under normal circumstances, alpha-fetoprotein (AFP) mainly comes from the liver cells of the embryo. About two weeks after the fetus is born, alpha-fetoprotein disappears from the blood. The serum alpha-fetoprotein of normal people is still less than 20ng / ml. During the period of liver cell regeneration, some embryonic carcinomas, germ cell tumors, digestive tract cancers, and AFP in the blood of pregnant women also increase to varying degrees. But the most noteworthy is that in patients with liver cancer, the concentration of AFP in the blood can be greatly increased,...

Examples

preparation example Construction

[0036] The preparation method of the test strip for rapid detection of alpha-fetoprotein in whole blood disclosed by the present invention comprises the following steps:

[0037] 1) Colloidal gold particles with uniform shape and size were prepared by the citrate reduction method of chloroauric acid, and the diameter of the particles was 20 nm.

[0038] 2) Prepare the colloidal gold immunoprobe, adjust its pH to 8.5-9.0, and modify the AFP monoclonal antibody 2 according to the principle that high-concentration salt solution can cause coagulation of unstable colloidal gold.

[0039] When modifying, mix 0.6-1.5ml of colloidal gold and 10-15μg of AFP monoclonal antibody 2 evenly and let stand at 4°C for 25-35 minutes. Add BSA for blocking for 35-45 minutes, keeping its final concentration at 0.5%-1.5%. Centrifuge at 10000-14000rpm under centrifugation conditions, and use PBS buffer containing 0.5%-1.5% BSA, 0.03%-0.1% sodium azide, 0.5%-1.5% sucrose, and 0.5%-1.5% Tween-20 for ...

Embodiment 1

[0044] 1) Preparation of 20nm colloidal gold: Take 100 mL of 0.01% (v / v) chloroauric acid solution, put it into a cleaned and dried Erlenmeyer flask, place it on a heating magnetic stirrer and heat it to boiling. Maintain the heating state and accurately and quickly add 2.0 ml of freshly prepared 1% (v / v) trisodium citrate solution in one go under vigorous stirring. It can be observed that the color of the solution changes from yellow→black→purple→dark blue→wine red, and continue stirring for 15 minutes until the solution turns transparent wine red. After the reaction solution was cooled to room temperature, it was stored at 4°C.

[0045] 2) Preparation of immune probe: first use 0.1M k 2 CO 3 The pH of the colloidal gold was adjusted to 8.7, and 13 μg of AFP monoclonal antibody 2 was added to 1 ml of the colloidal gold solution and left to stand at 4° C. for 25 minutes. 10% (v / v) BSA was added for blocking for 40 minutes, keeping its final concentration at 1%. Make the im...

Images