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A method for detecting hepatitis B surface antigen by inductively coupled plasma mass spectrometry

A technology of hepatitis B surface antigen and inductive coupling, applied in the field of mass spectrometry, can solve the problems of limited selection of markers, narrow detection range, radiation hazards, etc., and achieve the effect of excellent accuracy, rapid detection, and wide linear range

Inactive Publication Date: 2016-03-23
HIGH & NEW TECH RES CENT OF HENAN ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for samples with low HBsAg concentration, the sensitivity of detection is often low due to methodological reasons, and the detection range is narrow, resulting in missed detection in clinical practice, which brings losses to patients, especially patients who need clinical blood transfusion
Although other methods such as radioimmunoassay, chemiluminescence immunoassay, and fluorescence immunoassay have the characteristics of high sensitivity and wide linear range, their wider application is limited due to radiation hazards and limited selection of markers.

Method used

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  • A method for detecting hepatitis B surface antigen by inductively coupled plasma mass spectrometry
  • A method for detecting hepatitis B surface antigen by inductively coupled plasma mass spectrometry
  • A method for detecting hepatitis B surface antigen by inductively coupled plasma mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A kind of ICP-MS detection method based on the HBsAg of ZnSe colloidal solution labeling antibody, it comprises the steps:

[0023] 1) Preparation of ZnSe colloidal solution:

[0024] Preparation: refer to the literature [Enustun.B.V, Turkevich.J. J.Am.Chem.Soc .1963,85,3317-3328], specifically: Measure 125mL of double distilled water, fill it with nitrogen and remove oxygen for 1h, then add 0.875g (2.35mmol) ZnClO to the water under stirring conditions 4 ·6H 2 O and 5.7 mmol of MPA (used as a stabilizer), adjust the pH value of the reaction solution to 6.5 with 1 mol / L NaOH solution, and continue to pass nitrogen gas into the solution to remove oxygen. Under stirring, add 0.134g (0.46mmol) Al 2 Se 3 and excess sulfuric acid to react (to produce fresh H 2 Se 3 ), in this state, the precursor of ZnSe was produced. Then heat and reflux for 2 hours, after nucleation and growth stages, a ZnSe colloidal solution is obtained.

[0025] Purification: refer to literat...

Embodiment 2

[0034] A kind of ICP-MS detection method based on the HBsAg of nano-gold colloid solution labeling antibody, it comprises the steps:

[0035] 1) Preparation of nano-gold colloidal solution:

[0036] Preparation: Reference [B.Nikoobakht, M.A.El-Sayed, Chem.Mater., 2003,15,1957-1962,A.Gole,C.J.Murphy, Chem.Mate r., 2005, 17, 1325-1330] to synthesize a nano-gold colloidal solution through a seed-promoted growth process. The main process is: 5mL0.2mol / L cetyltrimethylammonium bromide (CTAB) and 5mL0.5mmol / L HAuCl 4 Mix, add 0.6mL ice NaBH with a concentration of 0.01mol / L under stirring 4 The solution was stirred vigorously for 2 minutes, and the solution was brownish yellow at this time, and then stood in a water bath at 25° C. for about 2 hours to obtain a gold seed solution.

[0037] Dissolve 3.4624gCTAB in 95mL of water, add 1mL of 0.01mol / L AgNO under stirring 3 solution, and then slowly add 0.6 mL of ascorbic acid (AA) solution with a concentration of 0.1 mol / L. Af...

Embodiment 3

[0046] Detection of clinical samples:

[0047] 10 serum samples were collected from the hospital, and tested according to the methods described in Examples 1 and 2, respectively. Calculate the corresponding concentration according to the linear equation, calculate the concentration of the antigen in the serum, and then compare it with the results obtained by the enzyme-linked immunoassay used in clinical diagnosis in the hospital. The results are shown in Table 1. In enzyme-linked immunoassay, if the normal value of HBsAg (ng / ml) is greater than or equal to 0.5ng / mL, it is regarded as a positive result (+), otherwise it is regarded as a negative result (-). It can be seen from Table 1 that the detection results obtained according to the methods of the two embodiments are basically consistent with the traditional clinical detection results, indicating that the detection method of the present invention can meet the clinical needs for HBsAg detection.

[0048]

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Abstract

The present invention provides a method for detecting hepatitis B surface antigen (HBsAg) by inductively coupled plasma mass spectrometry (ICP-MS), which involves covalently linking hepatitis B virus surface antibody (HBsAb) to metal colloid particles with activated carboxyl groups , the HBsAg in the serum and the colloidal particle-labeled HBsAb combine to form an immune complex through a specific reaction; the colloidal particles are dissociated from the immune complex under acidic conditions, and then the metal ion signal is detected by ICP-MS, thereby indirectly detecting HBsAg . Since each colloidal particle can be connected with one or more antibody molecules, the sensitivity of the detection is greatly improved; this method overcomes the disadvantages of low accuracy of the traditional colloidal gold immunochromatography and enzyme-linked immunoassay. It has excellent accuracy and high sensitivity, and the immunoassay detection is convenient and rapid, which broadens the application of ICP-MS in immunoassay.

Description

technical field [0001] The invention belongs to the field of mass spectrometry analysis, in particular to a method for detecting hepatitis B surface antigen by inductively coupled plasma mass spectrometry. Background technique [0002] China is a country with a high incidence of hepatitis B virus. According to epidemiological surveys, the incidence rate in the population is relatively high, and the surface antigen carrying rate is about 15%. The experimental diagnosis of hepatitis B virus has important reference significance for its treatment and prevention. Hepatitis B surface antigen (HBsAg) is the first serological marker to appear after HBV infection, and it is also one of the important indicators for diagnosis. For reasons such as testing costs, most domestic clinical laboratories use enzyme-linked immunosorbent assay (ELISA) to detect HBsAg. However, for samples with low HBsAg concentration, the detection sensitivity is often low due to methodological reasons, and t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/576G01N33/531G01N30/72
Inventor 刘珂珂刘冲庞海岩刘清黄海平段显英褚艳红
Owner HIGH & NEW TECH RES CENT OF HENAN ACAD OF SCI
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