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Kit for detecting amikacin content in blood and preparation method thereof

A technology of amikacin and a kit is applied in the field of latex-enhanced immune turbidimetric reagent and its preparation, which can solve the problems of troublesome sample processing, long time-consuming detection process, expensive detection equipment and the like, so as to improve detection sensitivity and enhance detection. Signal, the effect of good clinical application prospects

Inactive Publication Date: 2018-10-02
SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are not conducive to clinical application and promotion due to the expensive detection equipment, long detection process, troublesome sample pretreatment, and the need for full-time technicians.

Method used

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  • Kit for detecting amikacin content in blood and preparation method thereof
  • Kit for detecting amikacin content in blood and preparation method thereof
  • Kit for detecting amikacin content in blood and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Preparation of amikacin immunogen and anti-amikacin monoclonal antibody

[0044] Weigh 5.1 mg of amikacin and 55.0 mg of BSA and dissolve in 10 mL of PBS (phosphate buffer saline) (pH 7.2), stir well, add 20.0 mg of EDC dissolved in purified water dropwise, and stir at room temperature for reaction 6 Then the reaction solution was transferred to a dialysis bag, dialyzed in PBS (pH7.2) at 4°C for 4 days, centrifuged and then freeze-dried. Store at -20°C for later use. Referring to Yang Hanchun's "Animal Immunology" published in August 2003, Balb / C mice were immunized with amikacin-BSA (amikacin-bovine serum albumin) as an immunogen. According to the literature method (Zhu Liping, Chen Xueqing. Commonly used experimental methods in immunology. Beijing: People's Military Medical Publishing House, 2000), the monoclonal antibody was purified and obtained. The mouse Mab Isotyping Test Kit purchased from ROCKLAND Company was used to identify the subtype of the mon...

Embodiment 2

[0045] Embodiment 2: the preparation of chitosan and amikacin coupling conjugate

[0046] Dissolve 70mmol of chitosan in 10ml~30ml of water, then heat up the reaction system to 37~45°C; slowly add 70mmol of succinic acid dissolved in water dropwise, raise the temperature of the reaction system to 65°C, react for 4 hours, after the reaction, adjust the pH with to 7.0. Under the action of the activator EDC (carbodiimide), the above preparation is chemically coupled with the amikacin molecule to obtain the prepared conjugate;

Embodiment 3

[0047] Embodiment 3: the preparation of amikacin detection kit

[0048] 1. the kit of the present invention relates to the main raw material of reagent as follows:

[0049] anti-human amikacin antibody

[0050] Latex microspheres: use polystyrene latex particles with a diameter of 215nm and carboxyl groups (Agilent)

[0051] 2. the preparation of the main reagent of the present embodiment is as follows:

[0052] Reagent R1: 50mM PBS buffer, add 0.1% conjugate, 0.4% PEG6000 (polyethylene glycol 6000), 0.05% NaN 3 , 0.3% BSA, 0.8% NaCl, 0.2% Tween-20, pH 7.2, total preparation volume 1L, the reagent is a colorless or light yellow transparent solution.

[0053] Reagent R2: 50mM Tris buffer, latex microspheres coated with 0.2% amikacin antibody, 0.8% BSA, 0.9% NaCl, 0.05% NaN 3 , pH 7.4, the reagent is a milky white solution. Concrete preparation steps are as follows:

[0054] (1) 1.2g of 215nm latex microspheres, add 16ml of MES solution with pH 6.1, 2.5ml of freshly prepar...

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Abstract

The invention provides a kit for detecting amikacin content in blood. The kit is composed of a reagent R1 and a reagent R2 in a volume ratio of 1 to 1. The reagent R1 is prepared from a 20 to 150 mmol / L buffer, a 3 to 20 g / L conjugate, a 3 to 20 g / L stabilizer, a 5 to 20 g / L electrolyte, a 3 to 20 g / L sensitizer, a 0.5 to 10 g / L surfactant, a 0.2 to 1 g / L preservative, and purified water, and thepH is 7.0 to 9.5. R2 is prepared from 1.5 g / L to 3.5 g / L latex microspheres conjugated to an anti-human amikacin monoclonal antibody, a 20 mmol / L to 150 mmol / L buffer, a 3 g / L to 20 g / L stabilizer, a5 g / L to 20 g / L electrolyte, a 0.5 g / L preservative, and purified water, and the pH is 7.0 to 9.5. The kit provided by the invention is accurate, rapid and has a good clinical application prospect.

Description

technical field [0001] The invention relates to a detection reagent, in particular to a kit for detecting the content of amikacin in blood and a preparation method thereof, in particular to a latex-enhanced immune turbidimetric reagent which can be specifically used for clinically detecting the content of amikacin in blood and its preparation method. Background technique [0002] Amikacin, also known as amikacin, is an aminoglycoside antibiotic (Aminoglycosides), which is effective against most Enterobacteriaceae bacteria, such as Escherichia coli, Klebsiella, and Enterobacter , Proteus, Shigella, Salmonella, Citrobacter, Serratia, etc. all have good effects on Pseudomonas aeruginosa and other Pseudomonas, Acinetobacter, alkagenes Bacillus, etc. also have good effects; it also has good effects on Neisseria meningitidis, Neisseria gonorrhoeae, Haemophilus influenzae, Yersinia, Campylobacter fetalis, Mycobacterium tuberculosis and some non-tuberculous mycobacteria Antibacter...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/544G01N21/82
CPCG01N21/82G01N33/54346G01N33/544
Inventor 栾春杰张跃建孙卫兵
Owner SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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