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Homogeneous immunoassay kit for rapidly detecting procalcitonin and preparation method thereof, and detection method and device

A technology of procalcitonin and immunological reagents, applied in the field of immunoassay, can solve the problems of short shelf life, complicated washing steps, environmental pollution of radioimmunoassay, etc., and achieve the effect of rapid detection

Pending Publication Date: 2019-08-23
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing heterogeneous immunoassay methods require cumbersome washing steps; at the same time, it is necessary to overcome the defects of radioimmunoassay environmental pollution and short shelf life, and overcome the poor repeatability of enzyme immunoassay
[0004] There is no need for separation and washing steps in the homogeneous immunoassay technology. However, in the bioassay based on homogeneous immunoassay, when the detection efficiency is further improved by shortening the detection time, the performance of the detection will be seriously affected due to the unbalanced reaction, such as: The detection sensitivity will drop significantly
In addition, the short lifespan of singlet oxygen ions limits the application efficiency of photochemiluminescence technology, and there is no report on significantly increasing its lifespan, and increasing its lifespan can improve the detection signal under the same conditions, which can increase the detection sensitivity and reduce the detection rate. limit

Method used

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  • Homogeneous immunoassay kit for rapidly detecting procalcitonin and preparation method thereof, and detection method and device
  • Homogeneous immunoassay kit for rapidly detecting procalcitonin and preparation method thereof, and detection method and device
  • Homogeneous immunoassay kit for rapidly detecting procalcitonin and preparation method thereof, and detection method and device

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preparation example Construction

[0101] In some embodiments of the present invention, the acceptor is a polymer particle filled with a luminescent compound and a lanthanide element, and the preparation method of the first composition includes:

[0102] Step S1, the first antibody or its binding fragment is dialyzed with the first dialysis buffer to obtain the dialyzed first antibody or its binding fragment;

[0103] Step S2, after washing the receptor with a cross-linking buffer, resuspending the washed receptor with a cross-linking buffer to obtain a receptor liquid;

[0104] Step S3, adding the dialyzed primary antibody or its binding fragment to the receptor fluid and mixing evenly to perform binding to obtain receptors bound to the primary antibody or its binding fragment;

[0105] Step S4, after washing the receptor bound to the first antibody or its binding fragment with a washing buffer, adding a first buffer solution to obtain a first composition.

[0106] In some embodiments of the present invention...

Embodiment 1

[0181] Example 1: Preparation of a homogeneous immunoassay kit for rapid detection of procalcitonin

[0182] 1. Preparation of acceptor and donor

[0183] The preparation method, composition and content of the acceptor and donor used in the present invention can be referred to Example 1 in Chinese Patent CN100429197C (this patent document is hereby incorporated by reference in its entirety).

[0184] 2. Preparation of the first composition (R1)

[0185] (1) Pretreatment

[0186] Take 0.2 mg of PCT Ab1 to be treated and pack it into a dialysis bag (molecular weight cut-off is 14KD), put the dialysis bag into a beaker, add 100 times the volume of 0.05M CB buffer solution with pH9.6 in the beaker, and place it on a magnetic stirrer , Dialyze at 2-8°C, change the dialysate at least once, and dialyze for at least 4-5 hours each time to obtain PCT Ab1 after dialyzing, aspirate it and transfer it to a clean centrifuge tube, and take a sample to determine the protein concentration. ...

Embodiment 2

[0209] Implementation 2: Preparation of a homogeneous immunoassay kit for rapid detection of procalcitonin

[0210] Preparation of the first composition (R1)

[0211] According to the description in the examples of patent PCT / US2010 / 025433, PCT Ab1 is connected to the receptor to form a receptor that binds to PCTAb1. Wherein, the structure of the receptor is soluble, and it is in the form of non-particles, and the concentration of the receptor (component a) that binds to PCT Ab1 in the first composition (R1) is 200 μg / mL.

[0212] The preparation of all the other components is the same as in Example 1.

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Abstract

The invention relates to a homogeneous immunoassay kit for rapidly detecting procalcitonin and a preparation method thereof, and a detection method and device. The kit prepared by the method can rapidly detect procalcitonin in a to-be-detected sample, detection time can be shortened within 10 min, detection sensitivity can reach 20 pg / mL, and coefficient of variation is within 10%. When detectionis performed by adopting the kit, the using amount of a reagent can be decreased, raw materials can be saved, the content of the procalcitonin in the to-be-detected sample of a curee can be accuratelyand rapidly determined, and therefore, the kit has wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a homogeneous immune reagent kit for rapid detection of procalcitonin, a preparation method, a detection method and a device. Background technique [0002] Procalcitonin, namely procalcitonin (PCT), was discovered in 1990. It has been proved that its concentration in serum is closely related to the occurrence of infection. In normal individuals, its serum concentration is extremely low, only 10-50pg / ml, which cannot be detected by general methods. When systemic bacterial, fungal and parasitic infections, systemic inflammatory response syndrome, sepsis, acute and chronic pneumonia, acute pancreatitis, active hepatitis, trauma, etc. occur, the serum PCT level increases abnormally, and its concentration can reach normal levels several times to tens of thousands of times. When viral infection, autoimmune disease, organ transplant rejection, etc. occur, its seru...

Claims

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Application Information

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IPC IPC(8): G01N33/536G01N33/531G01N21/76
CPCG01N21/76G01N33/74G01N33/533G01N33/582G01N33/577G01N33/5306G01N2333/585G01N2800/7095G01N2800/26
Inventor 孙小禁杨阳洪琳赵卫国刘宇卉李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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