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Immunofluorescence kit for quantitatively detecting content of troponin I and preparation method

A troponin and immunofluorescence technology, which is applied in the field of quantitative detection of troponin I content immunofluorescence kits and preparation, can solve the problems of low sensitivity, inability to quantify, and poor repeatability, and achieve the effect of improving detection sensitivity

Inactive Publication Date: 2016-07-13
苏州联辰生物技术有限公司
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Problems solved by technology

[0004] The object of the present invention is to provide a kind of immunofluorescence reagent kit and preparation method for quantitatively detecting the content of troponin I, aiming at solving the problems of gel chromatography and high-performance liquid chromatography in the current method for detecting cTNI, which are time-consuming and difficult to automate; Poor accuracy, long operation time, low degree of automation, low sensitivity, generally only qualitative, not quantitative, especially the shortcoming of poor repeatability limits its clinical application, especially not suitable for accurate quantification to help treat Quantitative detection of body fluid marker proteins for disease diagnosis

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  • Immunofluorescence kit for quantitatively detecting content of troponin I and preparation method
  • Immunofluorescence kit for quantitatively detecting content of troponin I and preparation method
  • Immunofluorescence kit for quantitatively detecting content of troponin I and preparation method

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preparation example Construction

[0062] The preparation method of the antibody and particle complex comprises the following steps:

[0063] 1) Activation and washing of large particle latex particles, this step needs to activate the amino groups modified on the surface of large particles, centrifuge the commercialized latex latex solution after activation, discard the supernatant, and wash the precipitate repeatedly 3 times with washing buffer; The final precipitation is resuspended in the washing buffer, so that the mass volume ratio of the latex particles is 1% final concentration, and the washing buffer is selected from one of PBS buffer and MES buffer;

[0064] 2) Troponin I antibody coated latex particles

[0065] 1. Get 1ml of latex solution after activating and washing, add Troponin I antibody, make Troponin I antibody final concentration be 1mg / ml; After mixing, put 437 DEG C of shakers and coat with 220rpm rotating speed for 218h;

[0066] ②Wash the troponin I antibody not bound to the latex particl...

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Abstract

The invention discloses an immunofluorescence kit for quantitatively detecting the content of troponin I and a preparation method. The preparation method comprises the following steps: preparing a troponin antibody-large-particle fluorescent particle conjugate; preparing a nitrocellulose membrane labeled antibody; assembling a reagent card; preparing a TNI calibration product; establishing a standard curve; calculating the content of the troponin in a sample to be detected. The immunofluorescence kit for quantitatively detecting the content of the troponin I comprises a nitrocellulose membrane, a reagent and the calibration product. The lowest detection limit of the kit can reach 0.1ng / ml and the requirement of clinical application is met; the fastest time of a process from sample acquisition to detection result presentation only needs 10 to 15 minutes so that more expensive time for patients is provided, and illness states of the patients can be dynamically detected. The immunofluorescence kit for quantitatively detecting the content of the troponin I has good result relevance, no remarkable difference and an accurate and reliable detection result, and can be used for clinically replacing imported products; the detection cost is greatly reduced.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to an immunofluorescence kit for quantitative detection of troponin I content and a preparation method. Background technique [0002] Troponin is composed of three subunits of troponin T (TnT), troponin I (TnI) and troponin C (TnC), which are the main regulatory proteins of muscles. Because the three are tightly combined and calcium-dependent Sex, by the release of calcium to control muscle contraction function. Troponin I (TnI) is distributed in cardiomyocytes (cTnI) and mainly regulates the contraction of the myocardium. The concentration of cTnI in healthy people is lower than 20.4pg / ml. Because of its high specificity and high sensitivity, cardiac troponin I (cTnI) has become one of the highly sensitive and specific markers of myocardial injury, and it is also an important marker for judging myocardial injury, risk classification, and prognosis. Biochemical Indi...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/544
CPCG01N33/54313G01N33/544G01N33/68G01N2333/47
Inventor 翁濬一孙京海马雪林
Owner 苏州联辰生物技术有限公司
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