Method of determining ratio of globin chain alpha to globin chain beta in hemoglobin and application thereof

A technology of α-globin and β-globin, which is applied in the application field of detecting β-thalassemia, can solve the problems of complicated test operation, unfavorable screening popularization, high cost, etc., achieves broad application prospects, is convenient for long-term storage and long-distance Transportation, low cost effect

Active Publication Date: 2016-01-06
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, as a diagnosis after the onset, when hypochromic microcytic anemia is generally found through blood routine examination and iron deficiency anemia is ruled out, patients suspected of thalassemia are diagnosed

Method used

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  • Method of determining ratio of globin chain alpha to globin chain beta in hemoglobin and application thereof
  • Method of determining ratio of globin chain alpha to globin chain beta in hemoglobin and application thereof
  • Method of determining ratio of globin chain alpha to globin chain beta in hemoglobin and application thereof

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0048] The establishment of embodiment 1 detection method

[0049] (1) Preparation of reagents

[0050] Erythrocyte Hemoglobin Extractor (Reagent A): Deionized water; used for the extraction of erythrocyte hemoglobin.

[0051] Protein denaturant (reagent B): acetonitrile: formic acid solution (12g / L) in a volume ratio of 3:1; used for protein denaturation.

[0052] Protein lysing agent (reagent C): TPCK-Treated trypsin lyophilized powder is dissolved in 1 mol / L ammonium bicarbonate solution to prepare a lysing solution with a concentration of 5 g / L; it is used for protein hydrolysis.

[0053] Cleavage protein complex solvent (reagent D): acetonitrile aqueous solution (volume ratio of acetonitrile to water is 1:1), which also contains 1.2 g / L formic acid.

[0054] (2) Extraction and cracking of hemoglobin

[0055] ①Punch the dried blood filter paper sample with a standard hole punch to punch out a round filter paper blood sample with a diameter of 3.2mm (if the whole blood s...

Example Embodiment

[0109] Example 2

[0110] Using the above-established detection scheme, the liquid phase system flow injection method is used to quickly inject samples, and then the mass spectrometer is used for rapid detection to achieve high-throughput injection of detection samples. The detection time of each sample injection is less than 1 minute. In the experiment, at least 192 samples can be tested in the same batch.

[0111] According to the above technical scheme of the present invention, 200 normal controls (age 6 months to 40 years), 358 β-thalassemia patients and 16 HbE type anemia patients were analyzed, and the reference value interval (1st-99th) was established as shown in Table 10 . The reference interval may vary slightly between different laboratories, different experimenters, and different reagent batches. It is recommended that each laboratory establish its own reference value interval.

[0112] Table 10. Reference interval of α:β ratio in normal subjects, β-thalassemia c...

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Abstract

The invention belongs to the field of biological chemical detection and particularly relates to a method of determining the ratio of a globin chain alpha to a globin chain beta in hemoglobin and application of the method in detection of beta-mediterranean anemia. The method comprises the steps that hemoglobin samples are taken and split to obtain split fragments of the globin chain alpha and split fragments of the globin chain beta; one split fragment of the globin chain alpha and one split fragment of the globin chain beta are selected and used as sign peptide fragments; a mass spectrum is used for determining the concentration of the sign peptide fragment of the globin chain alpha and the concentration of the sign peptide fragment of the globin chain beta; the ratio of the globin chain alpha to the globin chain beta is expressed just by the ratio of the concentration of the sign peptide fragment of the globin chain alpha to the concentration of the sign peptide fragment of the globin chain beta. According to the method, by means of quantitative analysis of the ratio of the globin chain alpha to the globin chain beta, different types of beta-mediterranean anemia can be identified at an early stage, and severe, medium and mild beta-mediterranean anemia can be identified.

Description

technical field [0001] The invention belongs to the field of biochemical detection, and in particular relates to a method for measuring the ratio of hemoglobin alpha and beta globin chains and its application in detecting beta thalassemia. Background technique [0002] Hemoglobin (Hb) is composed of a globin and four heme (also known as ferrous protoporphyrin), and each heme is composed of four pyrrole subunits to form a ring, and the center of the ring is a ferrous ion. Each globin is composed of four polypeptide chains, and the four peptide chains of globin are different. Adults can be composed of 2 α chains and 2 β chains, called HbA, and fetuses are composed of 2 α chains and 2 γ chains, called HbF, which are replaced by HbA shortly after birth. Each polypeptide chain of globin is connected with a heme to form a monomer, or subunit (ie, subunit) of hemoglobin. [0003] Thalassemia, also known as thalassemia, is a common disease in children in my country, accounting for...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 余朝文邹琳
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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