Improved body fluid color-developing fungi 1,3-beta-D-glucan detection kit and application method thereof
A kit and dextran technology, applied in the direction of measuring devices, analyzing materials through chemical reactions, and analyzing materials through observing the influence of chemical indicators, can solve the problems of complex operation and low detection accuracy , to achieve the effect of simple production process, strong stability and small batch-to-batch difference
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Embodiment 1
[0037] Example 1: Production and manufacture of an improved human body fluid chromogenic fungal 1,3-β-D-glucan detection kit.
[0038] The improved human body fluid chromogenic fungal 1,3-β-D-glucan detection kit includes: main reaction agent, main agent complex solution, sample treatment solution, sterile water, standard and quality control products .
[0039] The preparation method of described reaction main agent comprises the following steps:
[0040] (1) Mixed batch of Limulus cell lysate: place the Limulus cell lysate at 4°C, thaw for 10 hours, mix at least 2.5kg of Limulus cell lysate, and fully stir for 20 minutes at 4°C;
[0041] (2) Emulsification of Limulus cell lysate: Homogenize the Limulus cell lysate after fully stirring in (1) with a homogenizer at 4 °C, the speed of the homogenizer is 10,000 rpm, and the homogenization time is controlled at 20 minutes Within, the limulus cell lysate is fully emulsified to obtain the limulus cell lysate emulsion;
[0042] (3...
Embodiment 2
[0050] Example 2: Application of an improved human body fluid chromogenic fungal 1,3-β-D-glucan detection kit to detect 1,3-β-D-glucan content in human blood samples.
[0051] The kit is obtained from Example 1. The implementation steps of the application of the kit include: making a standard curve, preparing a positive serum sample, processing the sample, adding the main reaction agent, and testing on the machine. The specific steps are as follows:
[0052] 1. Preparation of standard curve:
[0053] (1) Preparation of standard solution
[0054] Take 1 kit standard product, add sterile water to dissolve it, and prepare a 200pg / mL 1,3-β-D-glucan standard product solution, vortex and mix for not less than 30 seconds, and dilute it with sterile water gradient To 100pg / mL, 50pg / mL, 25pg / mL, 12.5pg / mL, 6.25pg / mL, 3.125pg / mL, use sterile water as blank control.
[0055] The kit has 5-fold dilution for sample processing, so when preparing the standard curve, the sample concentrati...
Embodiment 3
[0081] Example 3: Verification of the detection accuracy of an improved human body fluid chromogenic method fungal 1,3-β-D-glucan detection kit.
[0082] The implementation steps of the detection accuracy verification of the kit include: preparing test serum samples, testing on a machine, and statistically analyzing the results.
[0083] 1. Preparation of Test Serum Samples
[0084] (1) High value serum samples
[0085] The 1,3-β-D-glucan standard was dissolved in the serum of a healthy person, and a serum sample with a concentration of 500pg / mL was prepared for use as sample A.
[0086] (2) Low value serum samples
[0087] The 1,3-β-D-glucan standard was dissolved in the serum of a healthy person to prepare a serum sample with a concentration of 50pg / mL, which was sample B for later use.
[0088] (3) Mixed samples
[0089] Mix sample A and sample B at a volume ratio of 1:9, and set aside, that is, sample C.
[0090] 2. On-board testing
[0091]Use described kit to detec...
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