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Candida tropicalis PCR (polymerase chain reaction) assay kit and method

A detection kit and technology for Candida, which is applied in the field of microbial detection, can solve the problems of many influencing factors, poor repeatability, and low resolution, and achieve the effects of high sensitivity, short time-consuming, and simple operation

Active Publication Date: 2014-02-12
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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  • Candida tropicalis PCR (polymerase chain reaction) assay kit and method
  • Candida tropicalis PCR (polymerase chain reaction) assay kit and method
  • Candida tropicalis PCR (polymerase chain reaction) assay kit and method

Examples

Experimental program
Comparison scheme
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. (MgCl2 is very easy to absorb moisture, put it in a desiccator after using a 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 of ...

Embodiment 2

[0081] Embodiment 2 The use of kit of the present invention

[0082] 1. Reagent preparation:

[0083] 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.

[0084] 2. Centrifuge Taq DNA Polymerase and Uracil N-Glycosylase (UNG) briefly, and put them in a -20°C refrigerator for later use.

[0085] 3. After determining the number of reaction tubes n (number of samples + negative and positive controls), take out the CT-PCR reaction solution, and mix n×44.3μl CT-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.

[0086] 4. Transfer the extracted solid...

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Abstract

The invention relates to the field of microorganism assay, and in particular relates to a candida tropicalis PCR (polymerase chain reaction) assay kit. The kit comprises a PCR solution, wherein the PCR solution comprises primers and a fluorescent probe A; the primers are divided into a forward primer A and a reverse primer A; the nucleotide sequence of the forward primer A is shown in the SEQ ID NO.1; the nucleotide sequence of the reverse primer A is shown in the SEQ ID NO.2; and the nucleotide sequence of the fluorescent probe A is shown in the SEQ ID NO.3. The kit has the advantages of high sensitivity and specificity, stability, timeliness, convenience in operation and the like.

Description

technical field [0001] The invention relates to the field of microorganism detection, in particular to a PCR detection kit and a detection method for Candida tropicalis strains. 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 - Cryptococcales - Cryptococcalaceae. 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 human...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04G01N21/64
Inventor 何东华黄发平
Owner 泰普生物科学(中国)有限公司
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