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ELISA kit for severe patient disease evaluation and detection method thereof

A detection method and kit technology, applied in the field of immunoassay medicine, can solve the problems of low accuracy, long cycle, complicated operation, etc., and achieve good specificity and sensitivity, accurate disease severity assessment, simple and rapid early assessment. Effect

Inactive Publication Date: 2014-08-13
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] The purpose of the embodiments of the present invention is to provide an ELISA kit and its detection method for condition assessment of critically ill patients, aiming to solve the problems of complex operation, low accuracy and long cycle in the existing condition assessment methods

Method used

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  • ELISA kit for severe patient disease evaluation and detection method thereof
  • ELISA kit for severe patient disease evaluation and detection method thereof
  • ELISA kit for severe patient disease evaluation and detection method thereof

Examples

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experiment example 1

[0028] Experimental example 1, preparation of ELISA kit and detection method for condition assessment of critically ill patients

[0029] 1.1 Monoclonal antibody: use antibodies CHGA monoclonal antibody, product number ABIN933034, specification 100ul (1mg / ml), antibody type LGG1kappa;

[0030] 1.2 Coating: Coating formula, the concentration of CHGA monoclonal antibody is 10ng / ml, add 100ul coating solution to each well of the micro-reaction plate of the microplate, overnight at 4°C, wash 5 times with PBST washing solution;

[0031] 1.3 Blocking: add 100ul / well blocking solution, overnight at 4°C, wash with washing solution 5 times, store in refrigerator at 4°C;

[0032] 1.4 Measurement: Use the purchased CHGA standard product, double dilution, set up a blank control, and add the patient's plasma into different wells (100ul / well); after adding the secondary antibody and HRP enzyme-labeled secondary antibody in sequence, add the substrate chromogenic solution respectively , sto...

Embodiment 2

[0033] Example 2, a study on the correlation between the severity of illness and plasma CHGA concentration in critically ill patients.

[0034] 2.1 Methods: Using the above ELISA method, 20 healthy volunteers and 120 consecutive critically ill patients were tested for plasma CHGA within 24 hours after admission, PCT, CRP and other indicators were detected, and APACHEII and SOFA scores and 28-day mortality were recorded.

[0035] 2.2 Results: The plasma CHGA level of the healthy control group: 40.91±4.63ug / L, the plasma CHGA level of the septic shock group: 213.62±36.90ug / L, the CHGA concentration of the dead patients was significantly higher than that of the surviving patients [(190.51±19.26ug / L) vs(65.00±16.87ug / L)p<0.017], ROC analysis takes 139ug / L as the cutoff value, the sensitivity of CHGA to predict death is 80.0%, and the specificity is 81.6%.

[0036] Such as image 3 As shown, the results of ROC analysis show that: CHGA and APACHEII score have comparable ability to ...

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Abstract

The invention discloses an ELISA kit for severe patient disease evaluation and a detection method thereof. The detection method comprises the following steps: preparing a CHGA monoclonal antibody with a concentration of 10 ng / ml, adding 100 ul coating buffer in each reaction hole of an ELISA plate, leaving the ELISA plate to stand still for a night at a temperature of 4 DEG C, then washing the ELISA plate with a PBST washing liquid for five times; adding 100 ul confining liquid in each hole of the ELISA plate, leaving the ELISA plate to stand still for a night at a temperature of 4 DEG C, washing the ELISA plate with a washing liquid for five times, preserving the ELISA plate in a refrigerator at a temperature of 4 DEG C; using a bought CHGA standard substance, diluting in multiple proportions, setting a blank reference, respectively adding 100 ul / hole of the blank reference and 100 ul / hole of a patient plasma sample into different holes; orderly adding an enzyme-labeled second antibody and an HRP enzyme-labeled second antibody into the holes, respectively adding a substrate color-developing liquid, then adding a terminating liquid to stop color development, and finally measuring the OD value at 450 nm by an ELISA; wherein the ELISA kit for severe patient disease evaluation comprises an ELISA plate, a confining liquid container, an enzyme-labeled second antibody container, a substrate color-developing liquid container, and a terminating liquid container. The ELISA kit can simply and rapidly carry out early-stage evaluation on disease conditions of a severe patient, and has the advantages of very good specificity and sensitivity.

Description

technical field [0001] The invention belongs to the technical field of immunoanalysis medicine, and in particular relates to an ELISA kit and a detection method for evaluating the condition of critically ill patients. Background technique [0002] The etiology of severe patients is complex and their conditions are changeable. Although there are modern medical methods such as organ function support, their mortality rate is still high. Early and accurate assessment of the condition, timely and effective prevention and treatment are important ways to improve the prognosis of critically ill patients. How to make early and accurate disease assessment is a difficult problem for clinicians. [0003] At present, various critical illness scores commonly used in the ICU, such as APACHE and SOFA scores, mainly aim at the comprehensive evaluation of some important symptoms, signs, medical history, imaging and physiological parameters of the disease, and quantitative evaluation to achie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/535
CPCG01N33/577G01N33/53G01N33/535
Inventor 张丹陈晓迎
Owner CHONGQING MEDICAL UNIVERSITY
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