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An improved human body fluid chromogenic method fungal 1,3-β-d-glucan detection kit and application method thereof

A kit and dextran technology are applied in the direction of measuring devices, analyzing materials through chemical reactions, and analyzing materials through observing the influence of chemical indicators, which can solve the problems of low detection accuracy and complicated operation. , to achieve the effect of simple production process, strong stability and small batch-to-batch difference

Active Publication Date: 2018-10-23
KOCH BIOTECHNOLOGY(BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is complicated to operate and has low detection accuracy

Method used

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  • An improved human body fluid chromogenic method fungal 1,3-β-d-glucan detection kit and application method thereof
  • An improved human body fluid chromogenic method fungal 1,3-β-d-glucan detection kit and application method thereof
  • An improved human body fluid chromogenic method fungal 1,3-β-d-glucan detection kit and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

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: Use the homogenizer to homogenize the Limulus cell lysate after being fully stirred in (1) 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;

...

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: preparation of test serum samples, on-machine detection, and statistical analysis of the results.

[0083] 1. Preparation of Test Serum Samples

[0084] (1) High value serum samples

[0085] Dissolve the standard 1,3-β-D-glucan in the serum of a healthy person, prepare a serum sample with a concentration of 500pg / mL, and reserve it as sample A.

[0086] (2) Low value serum samples

[0087] Dissolve the standard 1,3-β-D-glucan in the serum of a healthy person, prepare a serum sample with a concentration of 50pg / mL, and reserve it as sample B.

[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 detect sample A, B,...

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Abstract

The invention relates to an improved body fluid color-developing fungi 1,3-beta-D-glucan detection kit, which comprises a reaction main-agent, a main agent combination solution, a sample treatment fluid, sterile water, a standard substance and a quality control material. With Tachypleus tridentatus Leach or Limulus polyphemus Linnaeus hemocyte lysate as a main raw material, the reaction main-agent contains G factor, B factor, C factor, clottable protein and a polypeptide chromogenic substrate. By a new preparation process, the reaction main-agent has characteristics of high stability and small intra-assay and inter-assay difference. The main agent combination solution adopts a new formula. After the main agent combination solution is mixed with the reaction main-agent, Tachypleus Amebocyte Lysate endotoxin reaction branch can be effectively shielded. According to the kit, rate-method enzyme kinetics detection is carried out by ELIASA. The detection speed is fast, and anti-interference performance is strong. In addition, the preparation technology is simple, and the product has stronger stability and higher specificity and sensitivity.

Description

technical field [0001] The invention relates to an improved human body fluid chromogenic method fungal 1,3-beta-D-glucan detection kit and application thereof. Background technique [0002] Deep fungal infections are also called invasive fungal infections (IFI). In the past two decades, with the application of a large number of broad-spectrum antibiotics, the development of bone marrow organ transplantation, the application of anticancer drugs, glucocorticoids and immunosuppressants, catheter interventional therapy, especially the prevalence of AIDS, like Candida The incidence of deep fungal infections such as hyperemia and systemic aspergillosis is gradually increasing, and the pathological features of patients with deep fungal infections are not obvious. The traditional diagnostic methods have a high false negative rate and a long diagnostic cycle. These factors make the diagnosis, prevention and treatment of systemic fungal infections very difficult, and the correspondin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/579G01N21/78
CPCG01N21/78G01N33/579
Inventor 不公告发明人
Owner KOCH BIOTECHNOLOGY(BEIJING) CO LTD
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