Time resolution fluorescent biosensor based on phosphorescent light emitting technology and application thereof

A time-resolved fluorescence and biosensor technology, applied in the field of immunoassays, can solve the problems of non-dispersive optical system spectral interference and the inability to obtain measurement results of elements, etc., and achieve the effect of sensitive qualitative and quantitative detection and analysis

Active Publication Date: 2013-01-09
武汉睿奇生物工程有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The purpose of the present invention is to solve the problem that the existing atomic fluorescence instruments are non-dispersive spectrometers. Due to the problem of spectral interference in the non-dispersive optical system, some elements cannot obtain accurate measurement results, and then provide a phosphorescence-based luminescence technology. Time-resolved fluorescent biosensors and their applications

Method used

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  • Time resolution fluorescent biosensor based on phosphorescent light emitting technology and application thereof
  • Time resolution fluorescent biosensor based on phosphorescent light emitting technology and application thereof
  • Time resolution fluorescent biosensor based on phosphorescent light emitting technology and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Quantitative detection of double-antibody sandwich method for detection of hepatitis B virus surface antigen HBsAg

[0049] 1. Drawing of standard working curve:

[0050] First, the purified HBsAg standard was diluted 1:10 with normal human serum (diluted with pH 7.20.02M PB buffer) as a diluent to prepare a series of concentration standards, the concentrations were: 0ng / ml, 10ng / ml, 25ng / ml 6 samples of ml, 50ng / ml, 100ng / ml, 200ng / ml. Secondly, each sample was tested 10 times with 10 HBsAg test strips, and the T value of the sample and the control C value read by the testing instrument for 10 times were respectively averaged, and finally the T value corresponding to each concentration was obtained according to the ratio of the two. / C results are listed in the table below. (Table 1)

[0051] Concentration (ng / ml)

[0052] Draw the standard working curve with the T / C value as the X coordinate and the HBsAg concentration as the Y coordinate. The ...

Embodiment 2

[0062] Example 2: Qualitative detection of HIV antibodies by double-antigen sandwich method

[0063] 1. Determination of the cut-off:

[0064] While measuring a large number of normal human serum samples, measure a considerable number of positive serum samples. If the measured value is normally distributed, then according to the characteristics of the μ test, the negative and positive cuts are firstly determined with a one-sided 99.5% confidence limit. -off value; if it is a non-normal distribution, the percentile method can be used to determine the cut-off value with one-sided 95% or 99%. After the cut-off value of the negative and positive population is determined, the cut-off value is determined according to the size of the "grey area" and considering the false positive and false negative rates in a comprehensive balance. The test result is positive if the measured value ≥ Cut-off value, otherwise it is negative.

[0065] 2. Actual test results:

[0066] The test strip o...

Embodiment 3

[0073] Embodiment 3: Quantitative detection competition method pattern detects morphine

[0074] 1. Drawing of standard working curve:

[0075] First, dilute the pure morphine standard with pH7.20.02M PB buffer to prepare a series of concentration standards, the concentrations are: 0ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 400ng / ml, 800ng / ml 6 samples. Secondly, each sample was tested 10 times with 10 morphine test strips, and the T value of the sample detected by the 10 times of detection equipment and the control C value were respectively averaged, and finally the T value corresponding to each concentration was obtained according to the ratio of the two. / C results are shown in the table below. (Table 4)

[0076] Concentration (ng / ml)

[0077] Take the T / C value as the X coordinate, and use the morphine concentration as the Y coordinate to draw the standard working curve. The expression of the standard working curve through statistical fitting is: Y=-11.406X+839.83, ...

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Abstract

The invention provides a time resolution fluorescent biosensor based on a phosphorescent light emitting technology and an application thereof. The time resolution fluorescent biosensor based on the phosphorescent light emitting technology comprises an exciting light path module, a photoelectric signal receiving and converting module, a control system module, an amplifying and shaping circuit, a coding interface circuit and a data cache module. The exciting light path module comprises a main detection exciting light module and an auxiliary optical scanning module. The photoelectric signal receiving and converting module comprises a main detection photoelectric conversion module and an auxiliary scanning photoelectric conversion module. According to the invention, a phosphorescent light emitting material (platinum/palladium porphyrin) is used as a biological marker, and the result is represented in form of an infrared optical signal under green light irradiation and can be interpreted by an instrument so as to quantitively detect a target object to be detected. According to the invention, a chromatographic test strip is divided into a sandwich method mode, a competition law mode, an indirect method mode and a capturing method mode to qualitatively and quantitively detect and analyze different objects to be detected in a sample quickly and sensitively according to different immunoreactions of the objects to be detected.

Description

technical field [0001] The invention relates to a time-resolved fluorescent biosensor based on phosphorescence technology and an application thereof, belonging to the technical field of immune detection. Background technique [0002] In order to solve the interference problem of FIA from samples, especially the strong and changing fluorescent background of biological samples, as well as scattering and other factors, a long-life, long-wavelength fluorescent label chromatography test strip (such as lanthanide metal chelate) has been greatly promoted. Compounds, porphyrin compounds) and the development of time-resolved fluorescence technology, the sensitivity of non-isotope labeling analysis has reached or exceeded the radioimmunoassay method. Phosphorescence analysis is the sister technology of fluorescence analysis. Compared with fluorescence, it has many unique advantages: (1) It has a large Stokes shift, the wavelength of phosphorescence is longer than that of fluorescence,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 谢爱武
Owner 武汉睿奇生物工程有限公司
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