Nucleic acid film strip and kit for identification of candida species and detection of gene mutation
A technology for bacterial species identification and nucleic acid film strips, which is applied in the direction of microorganism-based methods, microorganism measurement/inspection, biochemical equipment and methods, etc. It can solve problems such as high environmental requirements, fungal death, and long culture time, and achieve detection High sensitivity, accurate and reliable results, and short detection time
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Embodiment 1
[0044] Embodiment 1 The implementation of the concrete technical scheme of the present invention
[0045] 1. Methods for the identification of Candida species
[0046]According to clinical reports, there are seven main types of pathogenic Candida, namely, Candida albicans, Candida glabrata, Candida krusei, Candida tropicalis, Candida parapsilosis, Candida jiyemeng and Candida portuguese. More than 95%, therefore, this project selects seven kinds of Candida as the target strains for strain identification. According to the gene sequence difference of the insertion sequence ITS1 between the 18S rRNA and 5.8S rRNA of the seven Candida species, it was used as the basis for the identification of the strains. After gene sequence comparison and analysis, the insertion sequence ITS1 has certain differences in different Candida species, and it is feasible to be used as the basis for identification of Candida species.
[0047] 2. Method for detection of drug resistance of Candida albic...
Embodiment 2
[0063] Embodiment 2 The use of kit of the present invention
[0064] 1. Auxiliary reagents:
[0065] Solution A: 100mL 20×SSC, 10mL 10% SDS, add pure water to 1000mL, store at room temperature.
[0066] Solution B: 25mL 20×SSC, 10mL 10% SDS, add pure water to 1000mL, store at room temperature.
[0067] Solution C: add 100mL of 1M sodium citrate to 1000mL with pure water, and store at room temperature.
[0068] Chromogenic solution: add 1mL TMB and 2μL 30% H to 19mL solution C 2 o 2 .
[0069] 2. Sample processing
[0070] Method for extracting DNA from secretion samples: collect clinical vaginal secretion samples, and use a fungal DNA extraction kit to extract DNA from the samples.
[0071] 3. PCR amplification
[0072] Take out the PCR reaction solution containing the kit of the present invention, mark the tube wall, centrifuge at 5000 rpm for 2 seconds, and add 4 μL of the extracted sample DNA to be tested. Add 4 μL of positive quality control and negative quality co...
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