Alpha-thalassemia gene detecting kit

A technology for thalassemia and gene detection, applied in the biological field, can solve problems such as inconvenient use, harm, and false detection

Active Publication Date: 2016-07-27
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0019] The above products and methods all have the risk of missed detection or false detection of non-deletion type α-thalassemia or Thai type α-thalassemia, resulting in the birth of children with moderate to severe thalassemia, which brings a

Method used

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Experimental program
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Effect test

Embodiment 1

[0074] 1. Design and screening of primers and probes

[0075] The sequence of the probes on the membrane strip is as follows: figure 1 As shown, the result can be judged by the position of the spots.

[0076] According to the selected 4 kinds of deletion-type α-thalassaemia and 3 kinds of non-deletion-type α-thalassemia, design probe array (3x4), the meaning of each point on the film strip and its relationship with the normal control are shown in Table 2.

[0077] Table 2

[0078]

[0079] Note: figure 1 Each square in represents a probe, which is used to detect and diagnose different thalassemia genotypes. In Table 2, QSN, CSN and WSN were used as normal controls for the four deletion α-thalassemias. The last letter "N" stands for normal and "M" stands for mutant.

[0080] Obtain the α-globin gene sequence from the GenBank database, and design corresponding primers and probe sequences according to different types of deletion or mutation. Hybridization sensitivity and...

Embodiment 2

[0107] Instructions for use of the kit of the present invention:

[0108] 1. The main components of the kit are shown in Table 7

[0109] Table 7

[0110]

[0111] 2. Other main reagents (boxes) needed for this test

[0112] Whole blood genomic DNA extraction reagent: It is recommended to use the "Nucleic Acid Extraction Reagent" of Yaneng Biotechnology (Shenzhen) Co., Ltd. (record number: Yueshen Machinery No. 20150099; model: whole blood DNA (spin column type); specification: 25 people servings / box or 50 servings / box.)

[0113] 20×SSC: Dissolve 175.3g NaCl and 88.2g sodium citrate in 750mL pure water, adjust the pH value to pH7.0 with concentrated hydrochloric acid, and finally set the volume to 1000mL, and store under autoclaving. Store at room temperature.

[0114] 10% SDS: Dissolve 20g of SDS in 180mL of pure water, adjust the pH value to pH7.0 with 1N HCl, and finally adjust the volume to 200mL. Store at room temperature.

[0115] 1M sodium citrate: Dissolve 294...

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Abstract

The invention provides a kit capable of detecting four deletion alpha-thalassemia genes (-alpha<3.7>, -alpha<4.2>, --<SEA> and --<THAI>) and three non-deletion alpha-thalassemia genes (alpha<CS>alpha, alpha<QS>alpha and alpha<WS>alpha) at the same time.The kit comprises a DNA chip, a PCR solution I and a PCR solution II.The DNA chip comprises a substrate and probes fixed to the substrate.The kit has the advantages that based on the PCR-reversal point hybridization detecting principle, corresponding amplification primers and probes are designed according to the mutation or deletion loci of genotypes, biotin is used for labeling the primers, amidogen is used for labeling the probes, the DNA chip (nylon membrane) is adopted as a substrate, the probes are fixed to the nylon membrane, and PCR products amplified by the specific primers and the probes fixed to the DNA chip are subjected to hybridization and signal developing box interpretation to diagnose thalassemia.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for experimentally diagnosing seven α-thalassemia genes (including 4 deletion-type α-thalassemia genes and 3 non-deletion-type α-thalassemia genes) in clinical samples at one time . Background technique [0002] Thalassemia (referred to as "thalassemia") is a heritable hemolytic blood disease caused by a defect in the globin gene, which reduces or fails the synthesis of globin chains, resulting in an imbalance in the ratio of the globin chains that form hemoglobin. There are two main types: α-thalassemia and β-thalassemia. However, the carrier rate of α-thalassemia is much higher than that of β-thalassemia. [0003] α-thalassemia (including deletion-type α-thalassemia and non-deletion-type α-thalassemia) is one of the most common monogenic genetic diseases in the world. Guangxi, Guangdong, Yunnan, Hainan, Hong Kong and other provinces and cities in southern China and Its surro...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 刘福平郑瑜刘晶晶李印淑未纪涛
Owner 亚能生物技术(深圳)有限公司
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