High-sensitivity antenatal screening kit of Down's Syndrome in pregnant women at second trimester as well as preparation and detection methods of kit

A Down syndrome, high-sensitivity technology, applied in the field of prenatal screening kits, can solve the problems of limited sensitivity, long time-consuming, miscarriage, etc.

Active Publication Date: 2013-01-09
山东康华生物医疗科技股份有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Down syndrome, also known as Trisomy 21 or Down's Syndrome (DS), is the most common congenital defect that causes mental retardation. Its incidence rate is about 1 / 750, and its main features are severe mental retardation, with an IQ of 20 to 60, only 1 / 4 to 1 / 2 of normal people of the same age, small head, flat occiput, and thick nape. Symptoms such as small eye cleft, lateral upward tilt, and deep inner canthus are accidental and random, and the incidence will increase with the age of pregnant women. There is currently no effective treatment for this disease to prevent the occurrence of such diseases The method adopted is to carry out prenatal screening and necessary prenatal diagnosis during pregnancy, find out as early as possible and take corresponding measures. At present, the methods of clinical prenatal diagnosis mainly use chorionic villus sampling or amniocentesis for prenatal diagnosis , these methods have played a certain role in the prenatal detection of Down syndrome fetuses and the prevention of the birth of such fetuses. However, because amniocentesis is traumatic to a certain extent, it can cause miscarriage, and it is impossible to directly treat all fetuses on a large scale. Pregnant women are tested with a confirmed method, which also requires certain technical equipment conditions, takes a long time, and costs a lot, and it is impossible to carry out extensive testing among all pregnant women.
[0003]Now, the application of monoclonal antibody technology, enzyme labeling technology and enzyme-catalyzed chemiluminescence quantitative detection technology has also appeared to detect AFP and β-HCG in serum of pregnant women. For example, the Chinese invention patent with the application number "CN200610044608.X" titled "Down Syndrome Prenatal Screening Kit in the Second Trimester", however, has the disadvantages of complicated operations, complicated preparation methods, and limited sensitivity. limit

Method used

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  • High-sensitivity antenatal screening kit of Down's Syndrome in pregnant women at second trimester as well as preparation and detection methods of kit
  • High-sensitivity antenatal screening kit of Down's Syndrome in pregnant women at second trimester as well as preparation and detection methods of kit
  • High-sensitivity antenatal screening kit of Down's Syndrome in pregnant women at second trimester as well as preparation and detection methods of kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] A high-sensitivity prenatal screening kit for Down's syndrome in the second trimester, including a box body, and AFP-coated strips, AFP standard products, AFP quality control products, AFP enzyme conjugates, β -HCG coated strips, β-HCG standard, β-HCG quality control, β-HCG enzyme conjugate, sample diluent, luminescent substrate solution A, luminescent substrate solution B, concentrated washing solution.

[0058] The AFP-coated strip and the β-HCG-coated strip, preferably milky white opaque polystyrene 96-well chemiluminescent microplate plate coated strip; each well of the microplate is coated with solid-phase AFP Monoclonal antibody and β-HCG monoclonal antibody, the concentration of the above-mentioned monoclonal antibody is preferably 1ug / mL;

[0059] The AFP standard products are AFP standard solutions with concentrations of 0 ng / ml, 2.5 ng / ml, 5 ng / ml, 10 ng / ml, 20 ng / ml, and 40 ng / ml;

[0060] The β-HCG standard substance is a concentration of 0 mIU / ml, 2.5 mIU / ...

Embodiment 2

[0071] The preparation of the high-sensitivity prenatal screening reagent for Down's syndrome in the second trimester of the above-mentioned embodiment 1 comprises the following steps:

[0072] (1) Preparation of AFP, β-HCG monoclonal antibody-coated strips

[0073] aUsing monoclonal antibody technology, first use pure AFP or β-HCG to immunize mice according to routine, take splenocytes to fuse with mouse myeloma cells, screen and match, and establish AFP and β-HCG monoclonal antibody cell bank , and then inoculate the peritoneal cavity of mice to prepare ascites, and then purify the ascites to obtain AFP or β-HCG paired monoclonal antibodies with a specificity of 100% and a purity greater than 96%;

[0074] b Preparation of pre-coated plates: one end of the pre-coated AFP plate is painted yellow, and one end of the pre-coated β-HCG plate is painted red for distinction;

[0075] c Preparation of AFP coating solution: use 0.1mol / L carbonate buffer (recipe: 0.34g Na 2 CO 3 +...

Embodiment 3

[0103] The detection method of the prenatal screening kit for Down's syndrome in the second trimester of the above-mentioned embodiment 1 comprises the following steps:

[0104] (1) Sample collection

[0105] Take 3ml of venous blood from pregnant women in the second trimester (14-21 weeks), put it in a disposable blood collection tube, put it in a 37°C water bath for 20-30 minutes to fully coagulate the fibrinogen, centrifuge at 4000 rpm for 15 minutes, and absorb serum. If the serum sample cannot be detected in time, it should be frozen at -20°C. During the test, the sample should be thawed and mixed evenly, and repeated freezing and thawing should be avoided. It is best not to store serum samples in a refrigerator at 4°C for more than 3 days.

[0106] (2) Adding sample detection steps

[0107] AFP determination

[0108] a. Equilibrate reagents stored in a refrigerated environment to room temperature;

[0109] b. Take out the required microwell strips from the aluminum ...

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Abstract

The invention discloses a high-sensitivity antenatal screening kit of Down's Syndrome in pregnant women at second trimester as well as preparation and detection methods of the high-sensitivity antenatal screening kit. The kit comprises a kit body, AFP (Alpha Fetal Protein) and beta-HCG (Human Chorionic Gonadotropin) coated plates, AFP and beta-HCG standard products, AFP and beta-HCG quality control products, AFP and beta-HCG enzyme conjugates, sample diluent, light-emitting substrate liquids and concentrated scrubbing liquid. The preparation method comprises the following steps of: preparing monoclonal antibody coated plates of AFP and beta-HCG; preparing the standard products and the quality control products of AFP and beta-HCG; and preparing the enzyme conjugates of AFP and beta-HCG; and preparing the sample diluent, a light-emitting substrate liquid A, a light-emitting substrate liquid B, the concentrated scrubbing solution and the like. The high-sensitivity antenatal screening kit of the Down's Syndrome in the pregnant women at the second trimester has the advantages that the purpose of the antenatal screening of the Down's Syndrome in the pregnant women at the second trimester is realized, and the accordance rate with the clinic diagnosis reaches more than 99.7%; and the kit is high in sensitivity, strong in the specificity and accurate and reliable in result.

Description

technical field [0001] The invention relates to a prenatal screening kit, in particular to a high-sensitivity prenatal screening kit for Down syndrome in the second trimester of pregnancy and a preparation method thereof, belonging to the field of prenatal screening kits. Background technique [0002] Down's syndrome, also known as Trisomy 21 or Down's Syndrome (DS), is the most common congenital defect that causes mental retardation. Its incidence rate is about 1 / 750. Its main features are Severe mental retardation, the IQ is mostly 20-60, only 1 / 4-1 / 2 of the normal people of the same age, with symptoms such as small head, flat occiput, thick nape, small eye cleft, upward slant on the outside, deep inner canthus, etc. Its occurrence is accidental and random, and the incidence will increase with the age of pregnant women. There is currently no effective treatment for this disease. The way to prevent this kind of disease is to carry out prenatal screening and Necessary prena...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N33/531
Inventor 杨致亭杨爱香邱香廷宋玉翠
Owner 山东康华生物医疗科技股份有限公司
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