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
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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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