Zika virus fluorescent PCR detecting kit
A detection kit, Zika virus technology, applied in the direction of microbial determination/inspection, microorganisms, biochemical equipment and methods, etc., can solve the problem of no newborn baby, shorten the time required for detection, reduce workload, Guaranteed effect of specificity and accuracy
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Embodiment 1
[0022] Embodiment 1 A kind of Zika virus fluorescent PCR detection kit and its use
[0023] 1. Prepare a kit comprising the following components:
[0024] 1 tube of primer-probe mixture (25 μl / tube), 1 tube of RT-PCR reaction solution (125 μl / tube), 1 tube of RT-PCR enzyme system (75 μl / tube), 1 tube of positive quality control (200 μl / tube), Negative quality control (200μl / tube) 1 tube, DEPCH 2 O (2000μl / tube) 1 tube.
[0025] 2. Preparation of target gene recombinant plasmid
[0026] The entrusted company synthesized and purified the in vitro transcribed RNA of Zika virus NS1 gene, measured the concentration by UV spectrophotometer, then calculated the RNA copy number, and used DEPCH 2 O were diluted to 1.0 × 10 10 copies / mL~1.0×10 1 copies / mL of the standard.
[0027] 3. Real-time fluorescent quantitative PCR amplification and detection
[0028] 3.1 Reagent preparation:
[0029] Take the corresponding amount of primer-probe mixture 1 μl, RT-PCR reaction solution 5 μ...
Embodiment 2
[0038] Example 2 Application of a Zika virus fluorescent PCR detection kit to detect specific samples
[0039] Select dengue virus type 1, dengue virus type 2, dengue virus type 3, dengue virus type 4, chikungunya virus, fever with thrombocytopenia syndrome virus, Hantaan virus, Seoul virus, Puumala virus , Japanese encephalitis virus, chikungunya virus, Epstein-Barr virus, human cytomegalovirus, and measles virus, each positive sample was used as a specific sample. Nucleic acid was extracted from all samples, and the steps of PCR amplification and result analysis were implemented by reference Example 1 is carried out, and the detection of negative and positive quality control products is carried out at the same time.
[0040] Test result: the amplification curve of the negative quality control product is not S-shaped (see attached image 3 ), the amplification curve of the positive quality control product is an obvious S-shaped curve (see attached Figure 4 ), both negative...
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