Three Respiratory Pathogens (2019-nCoV Influenza A Influenza B) Nucleic Acid Test Kit (PCR-Fluorescent Probe Method)
This kit is used for the in vitro qualitative detection of novel coronavirus (2019-nCoV), influenza A virus and influenza B virus in human throat swab samples. The highly sensitive and specific nucleic acid detection can detect multiple viruses at one time, and the whole detection time is about 1 hour, which can achieve early detection, early isolation, early reporting and early treatment of respiratory tract infections, and provide accurate clinical diagnosis and infection early. It can provide accurate clinical diagnosis and infection control basis.
Three respiratory pathogens nucleic acid detection kit (PCR-fluorescent probe method) adopts TaqMan fluorescent probe method to design primer probes for fluorescent detection for the highly conserved and specific regions of 2019-nCoV, influenza A virus and influenza B virus, using different fluorescent groups for labeling, and the nucleic acids in the detection process are reverse transcribed into cDNA, and during the amplification process The specific primers and probes bind to the target sequences, and the formation of PCR products is fully synchronized with the accumulation of fluorescent signals through the DNA polymerase activity of Taq enzyme and 5'-3' exonuclease activity; the nucleic acids of 2019-nCoV, influenza A virus and influenza B virus in the samples are achieved by detecting the signals of different fluorescent moieties Qualitative detection of 2019-nCoV, influenza A virus and influenza B virus nucleic acids in samples by detecting different fluorescent group signals.
The kit contains an internal reference set as a quality control of the reagents, nucleic acid quality and the operation itself to avoid false negative test results.
1. Reagent preparation
1.1. Take out the reaction buffer and primer probe Mix from the kit, leave them at room temperature, wait for complete melting, shake and mix, centrifuge and set aside; take out the detection enzyme solution from the kit, centrifuge and set aside.
1.2. Prepare PCR-Mix according to the number of samples to be prepared (N=number of samples + 1 tube of positive control + 1 tube of negative control), dispense 20 μL of PCR-Mix per well into the fluorescence PCR octet tubes, and immediately freeze the reaction buffer, primer probe Mix and assay enzyme solution below -18℃ after use.
2. Sample processing
Samples were first inactivated using a water bath at 56°C for 30 min after receipt.
Extraction of collected pharyngeal swab samples should ensure that the RNA meets the amount of RNA required for the experiment, and the extracted RNA samples are immediately tested or stored below -70℃ (no more than 7 days); meanwhile, the corresponding volume of positive and negative quality control is taken for extraction.
3. Sample addition
Add 10 µL each of the negative RNA extracted in step 2, the positive RNA extracted in step 2, and the RNA of the sample to be tested into each reaction tube, cover tightly and centrifuge briefly.
4. Real-time amplification (see instructions for details)
4.1 Instrument channel and reaction volume selection
4.2 Real-time amplification conditions setting
【Test methods and test results are explained in detail in the manual】
|Detection principle||PCR-Fluorescent Probe Method||Applicable models||ABI 7500、SLAN-96P|
|Sample Type||Pharyngeal swabs||Package specification||50 tests/box|
|Test time||about 1 hours||Expiration date||9 mouths|
|Storage conditions||-18℃ away from light|