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A kind of whole blood pretreatment reagent and its application

A pretreatment agent and whole blood technology, applied in the field of clinical testing, can solve the problems of cumbersome operation, incomplete processing, and long processing time, and achieve the effects of reducing interference, improving repeatability, and ensuring accuracy

Active Publication Date: 2022-03-01
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Based on this, the purpose of the present invention is to provide a whole blood pretreatment reagent and its application to quickly process whole blood samples in order to solve the problems of long processing time, cumbersome operation and incomplete treatment of whole blood pretreatment reagents in the prior art , to achieve the purpose of rapid and accurate blood drug concentration detection

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  • A kind of whole blood pretreatment reagent and its application
  • A kind of whole blood pretreatment reagent and its application
  • A kind of whole blood pretreatment reagent and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1, the influence of different organic solvents

[0053] 1. Test material

[0054] Formula: 5v / v% ethylene glycol, 5mol / L zinc sulfate, different volumes of organic solvents, 0.9% Nacl solution to make up to 100v / v%.

[0055] Organic solvents: methanol, dimethyl sulfoxide, acetonitrile.

[0056] 2. Test methods and results

[0057] Take 300 μl of whole blood into a centrifuge tube, add more than 600 μl of whole blood pretreatment agent, mix well, let stand for 30 seconds, centrifuge at 14,000 rpm for 5 minutes, absorb the supernatant, and carry out cyclosporine A project on Antu A2000PLUS chemiluminescence instrument detection. The results are shown in Table 1.

[0058] Table 1 The influence of different organic solvents on the detection results

[0059]

[0060] The results showed that methanol had the weakest effect on the HRP detection system as a treatment agent, which could ensure a clear gradient in the detection of cyclosporin A in the treated w...

Embodiment 2

[0061] Embodiment 2, the impact of different surfactants

[0062] 1. Test material:

[0063] Control: the existing whole blood pretreatment agent-sample extract (purchased from Abbott Trading (Shanghai) Co., Ltd., item number 1L77.55).

[0064] Formula: 85v / v% methanol, 5v / v% ethylene glycol, 5mol / L zinc sulfate, 0.05v / v% different surfactants, 0.9% Nacl solution to make up to 100v / v%.

[0065] Surfactant:

[0066] Surfactant 1: NINATER411

[0067] Surfactant 2: BRIJ 35

[0068] Surfactant 3: IGEPALRCA210

[0069] Surfactant 4: TRITONRX-100

[0070] Surfactant 5: RHODASURFRON-870

[0071] Surfactant 6: ZonylRFSN100

[0072] Surfactant 7: CHEMALRLA-9

[0073] Surfactant 8: SURFACTANT 10G

[0074] 2. Test method:

[0075] Take 300 μl of whole blood into a centrifuge tube, add more than 600 μl of whole blood pretreatment agent, mix well, place it for 15-90 seconds, then centrifuge at 14,000 rpm for 5 minutes, absorb the supernatant, and measure the OD value on a spectro...

Embodiment 3

[0084] Example 3. Comparison of the effects of different whole blood pretreatment reagents and different detection methods on the detection of cyclosporine A. 1. Test materials:

[0085] Existing whole blood pretreatment agent - sample extract.

[0086] Whole blood pretreatment agent formula of the present invention: 85v / v% methanol, 5v / v% ethylene glycol, 5mol / L zinc sulfate, 0.05v / v% ZonylRFSN100, 0.9% Nacl solution to make up to 100v / v%.

[0087] 2. Test method:

[0088] Take 300 μl of whole blood into a centrifuge tube, add more than 600 μl of whole blood pretreatment agent, mix well, leave it for 30 seconds, centrifuge at 14,000 rpm for 5 minutes, absorb the supernatant, and use one-step method and time-lapse method on Antu A2000PLUS chemiluminescence instrument respectively. One-step method and two-step method were used to detect cyclosporine A items, and the results are shown in Table 4.

[0089] Table 4 The impact of different detection methods on the detection of cy...

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Abstract

The application belongs to the technical field of clinical detection, and discloses a whole blood pretreatment reagent and its application, especially the application in the immunological detection of blood drug concentration of cyclosporine A and tacrolimus. The whole blood pretreatment agent in the present invention includes organic solvent, divalent metal salt, surfactant and physiological saline, and the organic solvent is methanol and ethylene glycol. Compared with the prior art, the present invention has at least one of the following beneficial effects: 1) The whole blood pretreatment agent of the present invention can effectively reduce the interference brought by hemoglobin to the HRP detection system in the case of using an organic solvent, and ensure the accuracy of the detection result 2) For the first time, a surfactant with better hemolysis effect was used in the whole blood pretreatment agent to ensure that the whole process of single sample processing can be completed within 10 minutes; 3) Compared with the existing whole blood pretreatment agent, On the premise that the consistency of the processed samples with LC / MS remains unchanged, the repeatability is improved accordingly.

Description

technical field [0001] The invention belongs to the technical field of clinical detection, and in particular relates to a whole blood pretreatment reagent and its application, in particular to the application in the immunological detection of blood drug concentration of cyclosporine A and tacrolimus. Background technique [0002] Blood drug concentration monitoring is guided by the principles of pharmacokinetics to analyze and measure the concentration of drugs in the blood, to evaluate the curative effect or determine the dosing regimen, to make the dosing regimen individualized, to improve the level of drug treatment, to achieve clinical safety, Effective and rational medication. Blood drug concentration monitoring is usually used for drugs with narrow therapeutic window, strong toxicity, long medication cycle, and large individual differences after medication. [0003] Cyclosporine A (CsA) is a water-insoluble cyclic undecapeptide isolated from the spore Trichoderma in 1...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/535
CPCG01N33/54326G01N33/535
Inventor 朱宇皇韩晶王瑞李忠信渠海刘功成项立红陈小玲付光宇
Owner AUTOBIO DIAGNOSTICS CO LTD