Human pathogenic fungus immunofluorescence diagnostic reagent as well as preparation method and application thereof

A pathogenic fungus and immunofluorescence technology, which is used in measurement devices, instruments, scientific instruments, etc., can solve the problems of long test time, high instrument requirements, and complicated operation, and achieve short detection time, strong detection specificity, and stable properties. Effect

Inactive Publication Date: 2015-01-21
NANKAI UNIV
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Problems solved by technology

[0005] The purpose of the present invention is to provide an immunofluorescence diagnostic reagent for rapid detection of human pathogenic fungal infection and its preparation method in order to solve the problems of complex operation, high requirements for instruments, relatively long test time and high cost in the prior art.

Method used

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  • Human pathogenic fungus immunofluorescence diagnostic reagent as well as preparation method and application thereof
  • Human pathogenic fungus immunofluorescence diagnostic reagent as well as preparation method and application thereof
  • Human pathogenic fungus immunofluorescence diagnostic reagent as well as preparation method and application thereof

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Embodiment 1

[0026] (1) Isolation of strains: pathogenic fungal specimens collected from the sputum of patients infected with oral candidiasis in the laboratory. The standard strains were obtained by streak culture on CHROMagar medium plate for 48 h. For the morphological characteristics of the strain, see figure 1 . In the YPD medium, the bacterium was in the form of yeast; in the YPD medium supplemented with 10% serum, the cells transformed into the form of hyphae. Therefore, it was preliminarily confirmed that the strain was Candida albicans.

[0027] (2) Molecular biological identification of strains:

[0028] The genome of the strain was extracted by the glass bead shaking method; the 18S rDNA sequence was amplified from the strain genome by using fungal 18S rDNA universal amplification primers 18SF (5'-TACCTGGTTGATCCTGCCAGT-3') and 18SR (5'-CCTTGTTACGACTTTTTACTT-3') . The amplified sequence was cloned into the pMD19-T simple vector (Takara, China), and the transformant was dete...

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Abstract

The invention discloses a human pathogenic fungus immunofluorescence diagnostic reagent and a preparation method thereof. A polyclonal antibody or a monoclonal antibody is prepared from clinically-separated pathogenic fungi; then the antibody and a fluorescence nano quantum dot material are subjected to dehydration synthesis through amide to obtain a fluorescence nano quantum dot-antibody compound; the fluorescence nano quantum dot-antibody compound can be directly used for detecting a sample covered with an unknown antigen (a patient specimen to be detected) on a carrier material; and the carrier material can be coated with the antigen in advance and then the unknown antigen is detected. The human pathogenic fungus immunofluorescence diagnostic reagent has the advantages of strong specificity on the detection of clinical pathogenic fungus infection, good repeatability, convenience for operation, accuracy and rapidness in detection and the like; clinical medication can be effectively guided and the antibiotics abuse is reduced; and the diagnostic detection and treatment costs of patients are reduced so that good economic benefits and social benefits are generated.

Description

technical field [0001] The invention relates to a rapid detection immunofluorescence diagnostic reagent capable of detecting human fungal infection. It specifically relates to the preparation and application of immunofluorescence diagnostic reagents for human fungal infection. Background technique [0002] Pathogenic fungi are a very important class of clinical pathogenic microorganisms, mainly including Candida, Aspergillus, Penicillium, Cryptococcus, Trichophyton, and Sporotrichum. In healthy people, such fungi are usually in a symbiotic state with the body. However, when the body's immunity is reduced or local microecological disorders occur, this type of fungus can easily transform from a symbiotic state to a pathogenic state. In particular, opportunistic pathogenic fungi such as Candida, Aspergillus, and Cryptococcus can not only cause superficial infection, but also cause disseminated infection, which is life-threatening. In recent years, due to the continuous expan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/544
CPCG01N33/577
Inventor 李明春陈家童喻其林
Owner NANKAI UNIV
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