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Polymerase chain reaction (PCR) fluorescence detection kit and detection method for candida albicans

A Candida albicans and fluorescence detection technology, applied in the field of microbial detection, can solve the problems of poor repeatability, many influencing factors, low resolution, etc., and achieve the effects of short time consumption, simple operation, high sensitivity and specificity

Active Publication Date: 2012-04-18
泰普生物科学(中国)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] This method currently plays a dominant role in the monitoring, prevention and control of daily nosocomial infection, but it has many shortcomings such as low resolution, many influencing factors, and poor repeatability.

Method used

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  • Polymerase chain reaction (PCR) fluorescence detection kit and detection method for candida albicans
  • Polymerase chain reaction (PCR) fluorescence detection kit and detection method for candida albicans
  • Polymerase chain reaction (PCR) fluorescence detection kit and detection method for candida albicans

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Experimental program
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Effect test

Embodiment 1

[0033] The composition of embodiment 1 kit of the present invention

[0034] Main raw material sources and preparation methods:

[0035] Tris: analytically pure, a product from a qualified supplier, content 99.7%, infrared qualified, pH (5% water) 10.3-10.9, moisture 0.3%, melting point 167-171°C, qualified absorption system, qualified for the highest content of impurities .

[0036] MgCl 2 : Analytical pure, the product of a qualified supplier, the content is not less than 99%, the aqueous solution reaction is qualified, and the highest content of impurities is qualified. (MgCl 2 Very easy to absorb moisture, put it in a desiccator after using the new bottle).

[0037]EDTA: Analytical pure, the product of a qualified supplier, it is a white crystalline powder, soluble in water, the solution is acidic, hardly soluble in alcohol, the content is not less than 99.5%, the aqueous solution reaction is qualified, and the complexing force test is qualified. The highest content o...

Embodiment 2

[0079] Embodiment 2 The use of kit of the present invention

[0080] 1. Reagent preparation:

[0081] 1. Take out 10× concentrated cleaning solution A and 10× concentrated cleaning solution B, dilute them with sterile pure water at a ratio of 1:9 (volume ratio), and put them in a 4°C refrigerator for later use.

[0082] 2. Centrifuge Taq DNA Polymerase and Uracil N-Glycosylase (UNG) briefly, and put them in a -20°C refrigerator for later use.

[0083] 3. After determining the number of reaction tubes n (number of samples + negative and positive controls), take out the CA-PCR reaction solution, and mix n×44.3μl CA-PCR reaction solution, n×0.5μl Taq DNA Polymerase and n×0.2μl Add Uracil N-Glycosylase (UNG) into a centrifuge tube and oscillate to mix well. After centrifuging briefly, dispense 45 μl into each PCR reaction tube, cover the tube cap and transfer to the sample loading area, and store in a 4°C refrigerator in the dark. spare.

[0084] 4. Transfer the extracted solid...

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Abstract

The invention relates to the field of microbe detection, in particular to a polymerase chain reaction (PCR) fluorescence detection kit for candida albicans. According to the technical scheme, the fluorescence detection kit for the candida albicans comprises a PCR reaction solution; the PCR reaction solution comprises primers A and a fluorescence probe A; the primers are divided into an upstream primer A and a downstream primer A; and the nucleotide sequence of the upstream primer A is shown as SEQ ID NO.1, the nucleotide sequence of the downstream primer A is shown as SEQ ID NO.2, and the nucleotide sequence of the fluorescence probe A is shown as SEQ ID NO.3. The kit has the advantages of high sensitivity and specificity, stability, timeliness, convenience for operation and the like.

Description

technical field [0001] The invention relates to the field of microorganism detection, in particular to a PCR fluorescence detection kit for Candida albicans strains and a detection method thereof. Background technique [0002] In recent years, due to the widespread use of broad-spectrum antibiotics, the development of cancer chemotherapy, organ transplantation, and the increase in immunocompromised patients, especially AIDS patients, the chances of deep infection caused by fungi, especially Candida, are increasing. [0003] Candida belongs to the kingdom Fungi—Deuteromycota—Bacillus—Cryptococcus—Cryptococcus. It is a biphasic unicellular yeast. Candida is a budding yeast-like fungus, a typical opportunistic pathogen. The common Candida species known to be pathogenic are: Candida albican, Candida tropicalis, Candida.parapsilosis, Candida krusei, Candida stellatoidea, Ji Yemeng (Candida guilliermondii) and Candida glabrata (Candida.glabrata) and other eight species in the h...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/725
Inventor 何华东黄发平
Owner 泰普生物科学(中国)有限公司
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