Kit for detecting alpha-thalassemia genes

A technology for thalassemia and gene detection, applied in the biological field, can solve the problems of inconvenient use, harm, and missed detection of the kit

Active Publication Date: 2016-07-13
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

In addition, double heterozygosity for non-deleted α-thalassemia and deleted α-thalassemia can also lead to HbH hydrops fetalis syndrome
[0020] At present, there is no one that can quickly and easily detect six deletion types (-α 3 . 7 , -α 4 . 2

Method used

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  • Kit for detecting alpha-thalassemia genes
  • Kit for detecting alpha-thalassemia genes
  • Kit for detecting alpha-thalassemia genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Design and screening of primers and probes

[0055] 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.

[0056] According to the selected 6 kinds of deletion-type α-thalassaemia and 6 kinds of non-deletion-type α-thalassaemia, design the probe array (3x7), the significance of each point on the film strip and its relationship with the normal control are shown in Table 2.

[0057] Table 2

[0058]

[0059] Note: figure 1 Each square in represents a probe, which is used to detect and diagnose different thalassemia genotypes. The 6 deletion α-thalassemias in Table 2 were taken as normal controls together with QSN, CSN, WSN, 13N, 43-44N and 49N. The last letter "N" stands for normal and "M" stands for mutant.

[0060] Obtain the α-globin gene sequence from the GenBank database, and design corresponding primers and probe sequences according to different types of deletion or mutat...

Embodiment 2

[0088] Instructions for use of the kit of the present invention:

[0089] 1. The main components of the kit are shown in Table 7

[0090] Table 7

[0091]

[0092] 2. Other main reagents (boxes) needed for this test

[0093] 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.)

[0094] 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.

[0095] 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.

[0096] 1M sodium citrate: Dissolve 294...

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Abstract

The invention provides a kit capable of simultaneously detecting 6 deletion type alpha-thalassemia genes (-alpha<3.7>, -alpha<4.2>, --<SEA>, --<THAI>, --<FIL> and -alpha<27.6>) and 6 non-deletion type alpha-thalassemia genes (alpha<CS>alpha, alpha<QS>alpha, alpha<WS>alpha, alpha<13>alpha, alpha<43-44>alpha and alpha<49>alpha).The kit comprises a DNA chip, PCR reaction liquid I and PCR reaction liquid II, wherein the DNA chip comprises a substrate and probes fixed on the substrate.The kit has the advantages that the detection principle of PCR-reverse dot blot is used, the corresponding amplification primers and probes are designed according to the mutated or deleted sites of the genes, the probes are fixed on the DNA chip, and thalassemia is diagnosed through the hybridization of PCR products amplified by the specific primers and the probes fixed on the DNA chip and the interpretation of a signal developing box.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for one-time experimental diagnosis of 12 α-thalassemia genes (including 6 deletion-type α-thalassemia genes and 6 non-deletion-type α-thalassemia genes) in clinical samples . 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 surrounding...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6837C12Q1/686C12Q1/6883C12Q2600/156C12Q2600/158C12Q2565/501
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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