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Candida albicans enolase double-antibody sandwich ELISA detection method

The technology of Candida albicans enolase and Candida enolase is applied in the detection field of Candida albicans, and can solve the problems of inability to distinguish between filamentous fungi and yeast-like fungal infections, inability to distinguish specific types of infected Candida, and the like, To achieve the effect of high detection sensitivity

Pending Publication Date: 2021-10-12
中国人民解放军联勤保障部队第九八〇医院
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention is to overcome the existing defects and provide the Candida albicans enolase double-antibody sandwich ELISA detection method to solve the current clinically used antigen detection item for auxiliary diagnosis of IC as proposed in the background technology. Test, this detection method detects the common 1,3-β-D-glucan on the fungal cell wall, and its positive results cannot distinguish between filamentous fungi and yeast-like fungal infections, let alone distinguish specific infections within the genus Candida. category problem

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  • Candida albicans enolase double-antibody sandwich ELISA detection method

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Embodiment 1

[0044] Candida albicans enolase double-antibody sandwich ELISA detection method, comprising the following steps:

[0045] Step 1: Prepare a sufficient amount of Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, Candida quaternidae, Cryptococcus neoformans and Saccharomyces cerevisiae;

[0046] Step 2: Prepare main experimental instruments and fungal culture supernatant;

[0047] Step 3: Calculate the concentration of the mixed solution with a hemocytometer;

[0048] Step 4: Take 5×106 spores and add them to 10ml of culture solution containing 20% ​​calf serum, carry out shaking culture at 37°C and 200r / min, and take out 0.5ml of the culture solution every 12-24 hours, 8000 spores / g. Centrifuge at 4°C for 5 minutes. After separating the supernatant, store the supernatant at minus 20°C;

[0049] Step 5: Use the modified sodium periodate method to mark the price of HRP sheep anti-Eno;

[0050] Step 6: Use the goat anti-Eno labeled in the above step...

Embodiment 2

[0062] Candida albicans enolase double-antibody sandwich ELISA detection method, comprising the following steps:

[0063] Step 1: Prepare a sufficient amount of Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, Candida quaternidae, Cryptococcus neoformans and Saccharomyces cerevisiae;

[0064] Step 2: Prepare main experimental instruments and fungal culture supernatant;

[0065] Step 3: Calculate the concentration of the mixed solution with a hemocytometer;

[0066] Step 4: Take 5×106 spores and add them to 10ml of culture solution containing 20% ​​calf serum, carry out shaking culture at 37°C and 200r / min, and take out 0.5ml of the culture solution every 12-24 hours, 8000 spores / g. Centrifuge at 4°C for 5 minutes. After separating the supernatant, store the supernatant at minus 20°C;

[0067] Step 5: Use the modified sodium periodate method to mark the price of HRP sheep anti-Eno;

[0068] Step 6: Use the goat anti-Eno labeled in the above step...

Embodiment 3

[0080] Candida albicans enolase double-antibody sandwich ELISA detection method, comprising the following steps:

[0081] Step 1: Prepare a sufficient amount of Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, Candida quaternidae, Cryptococcus neoformans and Saccharomyces cerevisiae;

[0082] Step 2: Prepare main experimental instruments and fungal culture supernatant;

[0083] Step 3: Calculate the concentration of the mixed solution with a hemocytometer;

[0084] Step 4: Take 5×106 spores and add them to 10ml of culture solution containing 20% ​​calf serum, carry out shaking culture at 37°C and 200r / min, and take out 0.5ml of the culture solution every 12-24 hours, 8000 spores / g. Centrifuge at 4°C for 5 minutes. After separating the supernatant, store the supernatant at minus 20°C;

[0085] Step 5: Use the modified sodium periodate method to mark the price of HRP sheep anti-Eno;

[0086] Step 6: Use the goat anti-Eno labeled in the above step...

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Abstract

The invention discloses a Candida albicans enolase double-antibody sandwich ELISA detection method. The method comprises the following steps: 1, preparing sufficient Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, Candida guilliermondii, Cryptococcus neoformans and Saccharomyces cerevisiae; 2, preparing a main experimental instrument and a fungus culture supernatant; 3, calculating the concentration of the mixed solution by adopting a blood cell counting plate; and 4, adding 5 * 10 < 6 > spores into 10 ml of a culture solution containing 20% calf serum, carrying out shake culture at 37 DEG C and 200 r / min, taking out 0.5 ml of the culture solution every 12-24 hours, and centrifuging at 4 DEG C for 5 min. According to the invention, a double-antibody sandwich ELISA method is established by using the monoclonal antibody and polyclonal antibody aiming at the Candida albicans recombinant Eno, so that the natural Eno antigen generated in the proliferation process of the Candida albicans can be identified, through evaluation of performance indexes, the specificity and precision of the method can meet the requirements of clinical application, and the detection sensitivity is high.

Description

technical field [0001] The invention belongs to the technical field of detection of Candida albicans, in particular to a double-antibody sandwich ELISA detection method of Candida albicans enolase. Background technique [0002] Candida albicans is a parasitic bacterium that usually lives in the skin, mucous membranes, digestive tract and other organs of the human body. When the body's resistance decreases, Candida albicans will multiply. When the amount reaches a certain level, the human body will Disease, so Candida albicans is an opportunistic pathogen. Candida albicans can also be spread through public baths, tubs, bath towels, swimwear, clothing, medical devices, and dressings. [0003] Enzyme-Linked Immunosorbent Assay (ELISA) is an immunoassay method developed by Engvall and Perlmann of Stockholm University in Sweden in the 1970s. It is currently one of the most mature immunoassay methods for commercial application, and it is also widely used in medical experiments, ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/573G01N33/569G01N33/58
CPCG01N33/577G01N33/573G01N33/56961G01N33/581G01N2333/988
Inventor 贾克然王缚鲲雷达鑫贺政新
Owner 中国人民解放军联勤保障部队第九八〇医院
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