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Detection reagent for antiviral protein MxA and preparation method thereof

An anti-virus and protein technology, applied in the field of medical immunodiagnosis, can solve the problems affecting the test precision and operation error, and achieve the effect of shortening the test time, simplifying the operation procedure, and good applicability of the reagents.

Pending Publication Date: 2020-04-10
SUZHOU KANGHESHUN MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the instrument platforms commonly used in tertiary hospitals, the dilution and mixing operations can be completed by large-scale automatic biochemical analyzers, but on small semi-automatic platforms, especially POCT portable platforms (commonly used in hospitals below the third level and medical service units at the county and township level) The test performed requires the operator to perform it manually, which brings additional operation steps and introduces more operational errors, which affects the precision of the test.

Method used

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  • Detection reagent for antiviral protein MxA and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1) Preparation of latex reagent

[0030] There are differences between preparation methods 1 and 2 in the combination of surfactants.

[0031] Preparation method 1: Prepare activated latex particles: Take 0.18mL of carboxylated polystyrene microspheres with a particle size of 333nm and a concentration of 10mg / mL, add 3.6mL of 25mM MES with pH6.5, stir for 10min, and add 7.04mg of EDC activator powder After stirring at room temperature for 2.5 hours, the precipitate was collected by centrifugation at 12000 rpm, and the precipitate was resuspended in 1.8 mL of 25 mM MES with pH 6.5.

[0032] Preparation of sensitized latex particles: 0.18 mL of MxA polyclonal antibody with a concentration of 4.0 mg / mL was added to 1.8 mL of 25 mM MES at pH 6.5, and stirred for 1 min to prepare a solution of the antibody to be coupled. Quickly pour the activated latex particles in (1) into the antibody solution to be coupled and stir for 2.5 hours. The precipitate was collected by centri...

Embodiment 2

[0042] Reagent sensitivity (blank limit)

[0043] Using the reagents in Example 1, select the zero-value calibrator as a blank sample test, and test the concentration on the POCT device. The test was repeated 20 times, and the mean (X) and standard deviation (SD) of the 20 test results were calculated according to the standard curve established in Example 1, and X+2SD was calculated as the sensitivity of the reagent. The test results are shown in Table 1, showing that the sensitivity of the reagent obtained by Preparation Method 1 is 5 ng / mL, and the sensitivity of the reagent obtained by Preparation Method 2 is 15 ng / mL, indicating that the combination of surfactants can significantly improve the sensitivity of reagent detection . The clinical reference value for the detection of the antiviral protein MxA (MxA) is about 50ng / mL, and the sensitivity of the MxA latex-enhanced immune turbidimetric reagent prepared in Preparation Method 1 of the present invention is 10 times sma...

Embodiment 3

[0047] Reagent repeatability

[0048] With the single reagent test prepared in Example 1, the MxA concentration is the control substance of the normal value level of about 50ng / mL and the control substance of the abnormal value level of the MxA concentration of about 200ng / mL, each repeated test 10 times, calculate the measured value Mean (X) and standard deviation (SD). Calculate the coefficient of variation (CV) according to formula (1). The test results are shown in Table 2. According to the measurement results, the coefficients of variation CV=5.35% and 4.53% were calculated, which met the technical requirements of reagent CV<10%.

[0049] CV = (SD / X) × 100% ………………………(1)

[0050] In the formula:

[0051] CV - coefficient of variation;

[0052] SD - standard deviation;

[0053] X - the average value of the measured values.

[0054] Table 2. Repeatability measurement results of Example 1

[0055] Example 1 repeatability repeatability Assay number ...

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Abstract

The invention discloses a latex-enhanced immunoturbidimetric assay detection single reagent for antiviral protein MxA and a preparation method thereof. A single reagent is adopted, mixing is not needed, the sensitivity is high, the detection speed is high, various samples such as the whole blood, serum and plasma can be directly measured. Therefore, the reagent can be widely applied to various transmission or scattering analyzers including a common biochemical analyzer, a specific protein analyzer and the like.

Description

technical field [0001] The invention belongs to the field of medical immunodiagnosis, and in particular relates to the preparation and application of a latex-enhanced immune turbidimetric detection single reagent for antiviral protein MxA (MxA). Background technique [0002] Mx protein is a kind of protein with antiviral activity induced and regulated by interferon, which widely exists in vertebrates. It was first discovered in mice in 1963 and named Mx1, and its homologous protein was found in humans in 1985 and named MxA. Interferon is the first line of defense for the human body against viral infection. It can induce a large amount of expression of MxA after the human body is infected with the virus. Therefore, measuring the content of MxA in the human body can be used for the detection and diagnosis of viral infection. The serum concentration of MxA is generally below 50ng / mL in normal human body, and the concentration after virus infection is generally above 200ng / mL. ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N33/531G01N21/51G01N21/59
CPCG01N33/6866G01N33/54313G01N33/531G01N21/51G01N21/59
Inventor 刘向晖汪春芳陈胜胜
Owner SUZHOU KANGHESHUN MEDICAL TECH
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