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Kit for simultaneously detecting thalassemia mutant type and deletion type and application thereof

A thalassemia and deletion type technology, which is applied in the determination/testing of microorganisms, chemical libraries, combinatorial chemistry, etc., can solve the problems of missed detection and improvement of thalassemia detection technology, and achieves significant advantages, small detection range and low cost Effect

Pending Publication Date: 2018-11-13
BGI BIOTECH WUHAN CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented method allows for efficient amplification or identification of genes involved with sickle cell disease (SCD) through use of specific primers that are designed specifically against certain DNA sequences called beta globin proteins found on chromosomally normal red blood cells. These methods have been developed over several decades ago but they still require expensive equipment such as specialized instruments like flow cytometry machines.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the efficiency and effectiveness of early cancer identification tests used today. Current methodologies involve analyzing specific regions called alpha globin loci associated with certain diseases such as sickness syndrome, beta globulism, abnormal liver function, congenital hypothyroid disorder, lymphatic malaria, necrocytosis fugaxia, autochondrial recessed siderosis, fetal hyperchromasia, X-linked hypoplasmosis syndroma, tetanocyte anomalous ghost cell leukodystrophy, sporadic adhesion polyneuropathies, bovine hereditary dyshidase deficiency, porcini ceroids, pigment darkening skin lesions, rickler's syndrome and other similar conditions.

Method used

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  • Kit for simultaneously detecting thalassemia mutant type and deletion type and application thereof
  • Kit for simultaneously detecting thalassemia mutant type and deletion type and application thereof
  • Kit for simultaneously detecting thalassemia mutant type and deletion type and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0089] The inventors designed PCR primers for the α-globin gene and β-globin gene of thalassemia, and then performed multiple PCR combination screening on the obtained PCR primers, and designed matching primer tag sequences. details as follows:

[0090] 1. Primer design

[0091] (1) Design of mutant primers

[0092] First, bioinformatics analysis was performed on the sequences of HBA1, HBA2, and HBB genes (two α-globin genes and one β-globin gene, respectively), and the pseudogenes ψα1, ψα2, and β-globin with high sequence similarity to HBA genes were analyzed. Other genes in the protein gene cluster were included in the scope of bioinformatics analysis to find out the conserved and specific sequences of HBA1, HBA2, and HBB genes, and then design primers. The designed primer sequences were subjected to genome-wide comparison (blast) analysis. Except for the accurate alignment to the target HBA1, HBA2, and HBB genes, the primers without accurate alignment at other positions i...

Embodiment 2

[0144] Using the kit with 96 sets of index primers obtained in Example 1, referring to the above-mentioned "general method", the results after sanger sequencing are known (including 7 deletion types, no mutations, common mutations, rare mutations and newly discovered mutations ) of 95 samples were tested. details as follows:

[0145] 1. Sample extraction

[0146] DNA was extracted from 95 blood samples using a KingFisher automatic extractor, and each blood sample was from Chenzhou People's Hospital. The main extraction steps are as follows: Take out 3 deep-well plates and 1 shallow-well plate matched with the Kingfisher automatic extractor, add a certain amount of matching reagents according to the instructions and mark them, and place all the well-plates with reagents in the corresponding Select the program "Bioeasy_200ul Blood DNA_KF.msz" and press "star" to execute the program for nucleic acid extraction. After the program is finished, about 100ul of the eluted product i...

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Abstract

The invention discloses a kit for simultaneously detecting thalassemia mutant type and deletion type and application thereof, wherein the kit includes a plurality of amplimer groups for amplifying thalassemia genes; the amplimer groups include primers designed by aiming at alpha globin genes and beta globin gene, and mutant type detecting primers aiming at HBA1 genes, HBA2 genes and HBB genes, anddeletion type detection primers aiming at alpha thalassemia genes -alpha 3.7, -alpha4.2,--SEA,--THAI and beta deletion type SEA-HPFH, sinotype and Taiwantype. Through adopting the kit, the thalassemia mutant type and deletion type can be simultaneously detected, particularly, seven common deletion types and more than 300 mutant types can be simultaneously detected, and novel mutant types can be found out; furthermore, the cost is low, and the kit is much superior to the conventional thalassemia detection kit.

Description

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Claims

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Application Information

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Owner BGI BIOTECH WUHAN CO LTD
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