Alpha-thalassemia screening kit and application thereof in prenatal screening

A technology for thalassemia and prenatal screening, applied in the direction of microbial determination/examination, biochemical equipment and methods, etc., can solve the problems of invasive inspection, invasive operation, fetal impact, etc., achieving high accuracy and reducing Birth rates, effects of filling technological gaps

Active Publication Date: 2013-12-04
邯郸市康业生物科技有限公司
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Problems solved by technology

[0007] At present, the commonly used methods for the diagnosis of thalassemia include: Southern hybridization, multiplex Gap-PCR, multiplex ligase-dependent probe amplification (MLPA), PCR-oligonucleotide probe (ASO) method, real-time fluorescent quantitative PCR , dot hybridization, direct sequencing technology, etc. The above diag

Method used

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  • Alpha-thalassemia screening kit and application thereof in prenatal screening
  • Alpha-thalassemia screening kit and application thereof in prenatal screening
  • Alpha-thalassemia screening kit and application thereof in prenatal screening

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Embodiment

[0027] 10 mL of peripheral blood from 100 pregnant women were collected from 3 hospitals, and prenatal screening for α-thalassemia was carried out according to the following steps.

[0028] (1) Extraction of cell-free fetal DNA from peripheral blood of pregnant women;

[0029] ① Aseptically collect 10 mL of whole blood in 15 mL vacuum EDTA anticoagulant blood collection tubes to avoid hemolysis.

[0030] ②Centrifuge at 800g in a low-temperature centrifuge at 4°C for 10 minutes, and absorb the supernatant.

[0031] ③ Take the supernatant (about 4-5mL of plasma) and transfer it to a new 15 mL polypropylene test tube, centrifuge at 1600g, 4°C for 10 minutes.

[0032] ④ Take 1 mL of the supernatant (2.0 mL new polypropylene test tube), add 1 / 10 volume of proteinase K buffer (10X), pulse shake and mix for 30 seconds; then add proteinase K (proteinase K, 30∪ / mL, no DNase and RNase) to a final concentration of 0.1-1 mg / mL, pulse-shake and mix for 30 seconds.

[0033] ⑤Water at 60°...

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Abstract

The invention discloses an alpha-thalassemia screening kit and application thereof in prenatal screening and belongs to the technical field of prenatal screening of alpha-thalassemia. The alpha-thalassemia screening kit comprises a negative control sample, a positive control sample, PCRmasterMIX, and a PCR primer. Application of the alpha-thalassemia screening kit includes: amplifying fetal DNA in maternal peripheral blood by the designed primer so as to perform prenatal screening on alpha-thalassemia. The maternal peripheral blood is collected for prenatal screening of alpha-thalassemia, with no need for puncturing the amnion cavity and inserting pile tissue and with no injury to fetus, and the alpha-thalassemia screening kit is safe and reliable; accuracy is up to 99.99%; the technical blank of noninvasive prenatal screening of alpha-thalassemia is filled, and fewer children with diseases are born.

Description

technical field [0001] The present invention relates to prenatal screening for α-thalassemia. Background technique [0002] Thalassemia, also known as thalassemia, is a group of genetic diseases. Its pathogenesis is that the globin chain of synthetic hemoglobin is reduced or missing, which leads to abnormal structure of hemoglobin. The deformability of red blood cells containing abnormal hemoglobin is reduced, and the life span is shortened. It can be destroyed by the liver and spleen in advance, leading to anemia and even abnormal development. This disease is medically known as hemolytic anemia. [0003] α-thalassemia: The human α-globin gene cluster is located at 16Pter-p13.3, each chromosome has 2 α-globin genes, and a pair of chromosomes has a total of 4 α-globin genes. Most α-thalassemias (abbreviated as α-thalassemia) are caused by the deletion of the α-globin gene, and a few are caused by gene point mutations. If only one α gene on one chromosome is missing or defe...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王瑞晓
Owner 邯郸市康业生物科技有限公司
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