Real-time fluorescent quantitative PCR (polymerase chain reaction) kit for detecting candida glabrata
A Candida and fluorescence quantitative technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of time-consuming, expensive, complicated operation, etc., and achieve cost reduction, easy operation, and simple operation Effect
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Embodiment 1
[0039] Embodiment 1 Candida glabrata detection kit and its use
[0040] 1. Prepare a kit including the following components: 1 tube of primer-probe mixture (25 μl / tube), 1 tube of PCR reaction solution (250 μl / tube), 1 tube of PCR enzyme system (75 μl / tube), positive quality control product (200μl / tube) 1 tube, negative quality control product (200μl / tube) 1 tube, pure H 2 O (2000μl / tube) 1 tube. Positive quantitative standard 4 tubes (10 7 copies / ml, 10 6 copies / ml, 10 5 copies / ml, 10 4 copies / ml).
[0041] 2. Collection, preservation and transportation of specimens
[0042] 2.1 Specimen types: blood, sputum, alveolar lavage fluid, etc.
[0043] 2.2 Specimen collection, storage and transportation: Take 200ul of blood, sputum, alveolar lavage fluid, etc., put them in sterile centrifuge tubes, label them well, put them in a curling container and take them to the laboratory or at low temperature (-20℃~-70℃ ℃) frozen. Dry ice is used for long-distance transport of specim...
Embodiment 2
[0054] Example 2 Application of Candida glabrata detection kit to detect clinical samples
[0055] Select 3 cases identified as Candida glabrata negative through the culture method, 3 cases were identified as Candida glabrata positive specimens through the culture method, the nucleic acid extraction of the specimen, PCR amplification and result analysis steps were carried out with reference to Example 1, and at the same time Perform negative and positive quality control testing.
[0056] Test result: the amplification curve of the negative quality control product is not S-shaped (see figure 1 ), the amplification curve of the positive quality control product is an obvious S-shaped curve (see figure 2 ), both negative and positive quality control products meet the quality control requirements of the kit; the instrument draws a standard curve based on the positive quantitative standard (see image 3 ) has a good linear relationship, indicating that the amplification efficienc...
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