Kit for performing gene detection on middle east respiratory syndrome coronaviruses
A technology for respiratory syndrome and coronavirus, applied in the detection/testing of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve the problem of no cross-reactivity of respiratory viruses, and achieve the time required for detection The effect of shortening, reducing costs, and improving detection efficiency
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Embodiment 1
[0023] The preparation and use of embodiment 1 Middle East respiratory syndrome coronavirus gene detection kit
[0024] 1. Prepare a kit including the following components
[0025] 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 (200μ / l tube) 1 tube, DEPCH 2 O (2000μl / tube) 1 tube.
[0026] 2. Preparation of target gene recombinant plasmid
[0027] The entrusted company synthesized and purified the in vitro transcribed RNA of UPE and N genes of MERS-CoV, measured the concentration by UV spectrophotometer, and then calculated the RNA copy number, and used DEPCH 2 O were diluted to 1.0 × 10 5 copies / mL~1.0×10 2 copies / mL of the standard.
[0028] 3. Real-time fluorescent quantitative PCR amplification and detection
[0029] 3.1 Reagent preparation:
[0030] Take the corresponding amount of primer-probe mixt...
Embodiment 2
[0039] The detection of embodiment 2 specific sample
[0040] Select HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU13, H3N2 (influenza virus), H1N1 (influenza virus), Adenovirus (adenovirus) type5, RSV (respiratory syncytial virus), EV71 (enterovirus) , PIV-1 (parainfluenza virus-type 1), PIV-2 (parainfluenza virus-type 2), PIV-3 (parainfluenza virus-type 3), PIV-4 (parainfluenza virus-type 4), nasal Virus, HSV-1 (herpes simplex virus), Epstein-Barr virus, human cytomegalovirus, measles virus positive sample each 1 example is used as specific sample, nucleic acid extraction is carried out to all samples, PCR amplification and result analysis steps refer to embodiment 1 Carry out the detection of negative and positive quality control products at the same time.
[0041] Test result: the amplification curve of the negative quality control product is not S-shaped (see attached Figure 4 ), the amplification curve of the positive quality control product is an obvious S-shaped curve (s...
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