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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

Active Publication Date: 2015-01-07
杭州美联医学股份有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Defects of the above detection methods: the positive rate of direct microscopic examination or smear staining method is low, below 80%, which cannot meet the clinical requirements; at the same time, the above three methods cannot be used for bacterial identification, and cannot distinguish different Candida species the goal of
[0010] At present, there is no product that can simultaneously identify Candida strains and detect drug-resistant mutation genes of Candida albicans
At the same time, the existing Candida drug susceptibility and identification technology is still based on traditional fungal isolation culture and drug susceptibility test. During the cultivation process, the requirements for dispensing operation technology are high, and the environment requirements for fungal culture are high. A little careless operation may easily cause fungal infection. It is judged as a false negative due to death; and the culture time is long (about a week), which cannot meet the needs of rapid detection

Method used

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  • Nucleic acid film strip and kit for identification of candida species and detection of gene mutation
  • Nucleic acid film strip and kit for identification of candida species and detection of gene mutation
  • Nucleic acid film strip and kit for identification of candida species and detection of gene mutation

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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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Abstract

The invention relates to a diagnostic reagent of candida and particularly relates to a nucleic acid film strip and a kit capable of simultaneously performing identification of candida species and detection of gene mutation. The nucleic acid film strip for identification of candida species and detection of gene mutation comprises a substrate and nucleic acid probes immobilized on the substrate, wherein the nucleic acid probes comprise a nucleic acid probe used for identification of candida species and having base sequences shown in SEQ ID NO: 1-7, a nucleic acid probe used for detection of gene mutation of candida albicans and having base sequences shown in SEQ ID NO: 8-23 and an internal reference nucleic acid probe having a base sequence shown in SEQ ID NO: 24. The nucleic acid film strip and the kit are used for identification of the species and detection of drug-resistant mutant genes according to the gene sequences of candida, judging from molecular structure, results are accurate and reliable and the detection sensitivity is high; the detection time is short and the detection can be completed only just in eight hours.

Description

technical field [0001] The invention relates to a diagnostic reagent for candida, in particular to a nucleic acid membrane strip and a kit that can be used for candida strain identification and gene mutation detection. Background technique [0002] Vaginal candidiasis, also known as vulvovaginal candidiasis (VVC), is a common disease in obstetrics and gynecology. It is a reproductive system infection that is prevalent in women of childbearing age in all age groups. 75% of women have Suffered from fungal vaginitis at least once. [0003] Candida albicans (C.albicans) is the most common pathogen of candida vaginitis, accounting for about 70-90%. The same host can carry multiple kinds of Candida, and the same Candida of different genotypes can colonize the same or different parts of the host. In recent years, with the widespread use of antibiotics, the spectrum of pathogenic bacteria in candidal vaginitis has also changed. Cases of candida vaginitis caused by non-albicans su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/72
CPCC12Q1/6834C12Q2531/113C12Q2537/143C12Q2565/625
Inventor 裘惠良郑银娜邹耀东田洁
Owner 杭州美联医学股份有限公司
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