2019-Novel Coronavirus (2019-nCoV) RT-PCR Detection Kit
Detection Principle and Performance Evaluation
TaqMan real-time PCR technology, FAM channel detect ORF1ab gene, JOE channel detect N gene, ROX channel detect E gene, CY5 channel detect internal control
- . Limit of detection: 300 copies/mL
- . Precision: CV of CT value within / between batches is less than 5%
- . Specificity: No cross reaction of other related pathogens with the same or similar infection site
RT-PCR Mixture preparation :
- • Use ice box to prepare reagent, and prepare reaction solution according to the number of reaction samples (number of reaction samples = number of samples to be tested + 2 control samples + 1)n.
- • Add RT-PCR enzyme n × 6 μL and 2019-nCoV reaction solution n × 14 μ l into the centrifuge tube, shake and mix, and centrifuge at low speed for several seconds.
- • The reaction tube is divided into 20 μL per tube and can be placed at 2-8 ℃ for 3 hours.
- • Nucleic acid extract adding: add 10 μL of nucleic acid extract into the reaction tube respectively, centrifuge at low speed for several seconds, and place it on the real-time fluorescence quantitative PCR instrument.
Nucleic Acid Extraction :
- • It is recommended to use the nucleic acid extraction and purification reagent (universal) produced by our company, QIAamp Viral RNA Mini Kit of QIAGEN, and NX-48 Viral RNA Kit of Genolution for extraction.
- • The required sample volume is 200 μL, and 5 μL internal control is added to each sample to be extracted (including the controls).
- • After extraction, the nucleic acid extract should be added to the reaction tube within 10 minutes, or transferred to the centrifuge tube and stored at - 15 ℃ - 25 ℃.
Product Advantages :
- • 3 target detection: qualitative detection of ORF1ab gene, N gene and E gene, reduce recheck and improve detection efficiency
- • Full-process internal monitoring: internal control were used to effectively monitor the whole detection process of extraction, reverse transcription and amplification, to avoid false negative results
- • Anti-contamination system: UNG enzyme and dUTP were used to effectively prevent the contamination of PCR amplicon, to avoid false positive results
- • Rapid Turnaround Time: with rapid nucleic acid extractor and extraction reagent, the nucleic acid can be extracted and purified quickly and safely, and the detection of 96 samples can be completed in 2 hours