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Nucleic acid film tape and kit for diagnosing alpha mediterranean anemia

A technology for thalassemia and nucleic acid membrane strips is applied in the field of diagnostic reagents for thalassemia, which can solve the problems of missed diagnosis, inability to detect alpha gene mutation, EB pollution, etc., and achieve the effect of reducing economic burden, family burden and social cost.

Active Publication Date: 2007-12-26
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

The disadvantage of this method is that it can only detect the deletion of the α gene, but not the mutation of the α gene, which can easily lead to missed diagnosis and result in the birth of defective babies
The technical solution is to use PCR technology to amplify the α-globin gene, and analyze whether the α-globin gene is missing or mutated by electrophoresis. This method is likely to cause EB pollution
[0014] So far, there are no published patents or literature reports for α-thalassemia-based nucleic acid hybridization membrane strips based on nylon membranes and mutated genes located in the middle of oligonucleotide probes

Method used

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  • Nucleic acid film tape and kit for diagnosing alpha mediterranean anemia
  • Nucleic acid film tape and kit for diagnosing alpha mediterranean anemia
  • Nucleic acid film tape and kit for diagnosing alpha mediterranean anemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0100] Embodiment 2 is used for the design of the primer of DNA in the sample to be amplified

[0101] According to the α-thalassemia gene sequence data, 4 pairs (a total of 7, one shared by two pairs) were used to amplify α - a globin gene fragment. The nucleotide sequences of these seven primers are shown in SEQ ID NO.16-22. The amplified product of SEQ ID NO.16 and SEQ ID NO.22 can be used with-- SEA Probe hybridization, used to detect the presence of -- SEA Deletion; the amplification products of SEQ ID NO.17 and SEQ ID NO.20 can be combined with -α 4.2 Probe hybridization, used to detect the presence of -α in the sample 4.2 Deletion; the amplification products of SEQ ID NO.18 and SEQ ID NO.21 can be combined with -α 3.7 Probe hybridization, used to detect the presence of -α in the sample 3.7 Deletion; the amplified products of SEQ ID NO.18 and SEQ ID NO.19 can be hybridized with the αα probe to detect whether the two chromosome groups of the sample are missing (if t...

Embodiment 3

[0109] The detection of embodiment 3 to-be-checked sample DNA

[0110] 1. Experimental instruments and reagents

[0111] 1. Test equipment

[0112] PCR instrument, DNA electrophoresis instrument, molecular hybridization box, shaker

[0113] 2. Test reagents

[0114] (1) 3% H2O2

[0115] (2) 20×SSC (pH7.0): Take 175.3g of NaCl and 88.2g of sodium citrate, add H2O to dissolve to 1000mL, adjust the pH to 7.0 with a pH meter, and autoclave.

[0116] (3) 10% SDS (pH7.0): Dissolve 20g of SDS in 180mL of H2O, adjust the pH to 7.0 with HCl, and finally set the volume to 200mL.

[0117] (4) Liquid A (2×SSC, 0.1% SDS, pH7.4): Take 100mL of 20×SSC, 10mL of 10% SDS,

[0118] Add H2O to dissolve to 1000mL, and adjust the pH to 7.4.

[0119] (5) Solution B (0.5×SSC, 0.1% SDS, pH7.4): Take 25mL of 20×SSC, 10% SDS 10mL, add H2O to dissolve to 1000mL, and adjust the pH to 7.4.

[0120] (6) Solution C (0.1M sodium citrate, pH5.4): Dissolve 14.7g of sodium citrate in 450mL of water, adjus...

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Abstract

This invention relates to test kit and test paper with oligonucleotide probes for diagnosing alpha-thalassemia. The test paper comprises a base, and specific oligonucleotide probes immobilized on the base. The oligonucleotide probes comprise: specific oligonucleotide probes for detecting mutation of alpha-globin gene, specific oligonucleotide probes for detecting deletion of alpha-globin gene, and base sequence complementary with the above sequences. The test kit comprises specific primer for amplifying alpha-globin gene or transcript, and specific oligonucleotide probes for detecting mutation / deletion of alpha-globin gene. The test kit and the test paper have such advantages as low cost, rapid detection, and stable results. The test kit and the test paper can be used in detection of mutation / deletion of alpha-globin gene, and prenatal diagnosis of severe thalassemia (alphaTalpha) caused by point mutation, and do not have error diagnosis.

Description

technical field [0001] The invention relates to a diagnostic reagent for thalassemia, in particular to a nucleic acid membrane strip and a kit for diagnosing α-thalassemia. Background technique [0002] Thalassemia (thalassemia, hereinafter referred to as thalassemia) is due to the decrease in the synthesis rate of one or some globin chains in the patient, resulting in the lack of some peptide chains and the relative excess of other peptide chains, resulting in an imbalance in the number of peptide chains, resulting in hemolysis sexual anemia. There are two main types of thalassemia, α-thalassemia caused by a deficiency of α-globin chain synthesis and β-thalassemia caused by a deficiency of β-globin synthesis. [0003] Alpha-thalassemia is one of the most common monogenic genetic diseases in the world. The disease occurs in a wide range from Italy, Greece, Malta, Cyprus along the Mediterranean Sea to Southeast Asian countries. Guangxi, Guangdong, Hainan, Hong Kong, Taiwan...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor LIAO SHENGYUNTIAN JIEREN WEIXIANG ZHUZOU YAODONG
Owner 亚能生物技术(深圳)有限公司
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