The six respiratory pathogens nucleic acid detection kit (PCR-fluorescent probe method)
This kit is used for the in vitro qualitative detection of influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, rhinovirus and Mycoplasma pneumoniae nucleic acid in human throat swab samples, with high sensitivity and specificity. It can provide accurate clinical diagnosis and infection prevention and control basis.
The six respiratory pathogens nucleic acid detection kit (PCR-fluorescent probe method) uses Taqman fluorescent probe method to design primer probes for fluorescent detection for highly conserved and specific regions of influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, rhinovirus and Mycoplasma pneumoniae, using different fluorescent moieties for labeling, the nucleic acid during detection is reverse transcribed into cDNA, during amplification, the specific primers and probes bind to the target sequence, 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 detection of signals of different fluorescent moieties is achieved for the detection of influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, and mycoplasma pneumoniae in the sample. The kit is designed for the qualitative detection of influenza A virus, influenza B virus, adenovirus, rhinovirus and Mycoplasma pneumoniae nucleic acids in samples by detecting different fluorescent groups.
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 assay 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|