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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 by combining hemoglobin electrophoresis and genetic diagnosis techniques. Complicated, expensive, and detrimental to universal screening
Currently, there is no rapid and high-throughput detection method for thalassemia

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

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The establishment of embodiment 1 detection method

[0049] (1) Preparation of reagents

[0050] Erythrocyte hemoglobin extraction agent (reagent A): deionized water; used for extraction of erythrocyte hemoglobin.

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

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

[0053] Lysed protein reconstitution 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] ①Use a standard puncher to punch out the dried blood filter paper sample with a diameter of 3.2 mm circular filter paper blood sheet (if the whole blood sample is taken directly f...

Embodiment 2

[0110] Utilize the detection scheme established above, quickly inject samples through the flow sampling method of the liquid phase system, and then perform rapid detection through the mass spectrometer, so as to realize high-throughput sample injection of test samples. The injection detection time of each sample is less than 1 minute, once In the experiment, at least 192 samples can be tested in the same batch.

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

[0112] Table 10. Normal people, β-thalassemia carriers, and p...

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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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