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Product Details
| Cat # +Size | K1461-100 |
|---|---|
| Size | 100 rxns |
| Detection Method | qPCR based detection kit |
| Applications | • An ideal tool to detect Coronovirus SARS-CoV-2 by RT-PCR method |
| Features & Benefits | • Compatible with standard RT-qPCR machines (Bio-Rad CFX96, QuantStudio’s 7 Flex system)
• Highly specific for the RdRp and N target markers recommended by WHO and US CDC • Reliable and Ready-to-use • Includes a Positive Control • Results ready in less than 2 hr |
| Kit Components | • COVID-19 Primers/Probes
• 2X RT-qPCR Master Mix • Positive Control Template • Negative Extraction Control • RT-qPCR Enzyme Mix 40 μl • Nuclease-free Water 1 ml |
| Storage Conditions | -20ºC |
| Shipping Conditions | Dry Ice |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
BioVision’s Coronavirus rapid RT-qPCR Detection Kit is used for the detection of SARS-CoV-2 in respiratory specimens using Reverse Transcriptase Quantitative Polymerase Chain Reaction (RT-qPCR). It allows the efficient cDNA synthesis and qPCR in a single tube. This kit includes a 2X RT-qPCR Mastermix that contains all the reagents supplied in a 2X concentration to perform the qPCR. A separate RT-qPCR Enzyme Mix for cDNA synthesis is included the kit. Additionally, the kit contains 2 vials of primers and probes specific for the ORF1ab and N target. The PCR Detection Kit will facilitate the Coronavirus research and improve patient treatment.
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What are the qPCR Thermal Cyclers that are compatible with this kit?
The qPCR Thermal Cyclers that are compatible with this kit include Agilent MX3005P, Agilent MX4000, Agilent AriaMx, BioRad CFX96, BioRad iCycler iQ, Cepheid Smartcycler II, Illumina Eco, Qiagen Rotor Gene Q 5plex, Roche LightCycler 480 and Roche Cobas z 480.
What are some of the performance characteristics and QC data that you can share with us regarding K1461? Have you tested this kit for cross reactivity?
Clinical Performance: A study was conducted to evaluate the clinical performance of the kit. A total of 60 clinical specimens (30 negative and 30 contrived positive specimens) were tested. Results showed 100% Positive Percent Agreement (30/30 positive results) and 100% Negative Percent Agreement (30/30 negative results). However, K1461 is recommended for research use only.
Cross-Reactivity: The detection results of this kit displayed no cross-reactivity with influenza A virus, influenza B virus, adenovirus, Staphylococcus epidermis, HCoV-SARS, HCoV-OC43, HCoV-HKU1, or Streptococcus pneumonia.
Cross-Reactivity: The detection results of this kit displayed no cross-reactivity with influenza A virus, influenza B virus, adenovirus, Staphylococcus epidermis, HCoV-SARS, HCoV-OC43, HCoV-HKU1, or Streptococcus pneumonia.
What does the abbreviation RP in the kit stand for?
RP stands for the Ribonuclease P (RNase P) gene. The RP probes/primers provided in the kit are used to make sure that the samples showing negative results for COVID-19 contain nucleic acid for testing. RP serves to assess the nucleic acid extraction procedure.
What are the primers and probes provided in the kit?
The kit component (K1461-100-1) contains primer/probes for RNA-dependent RNA polymerase (RdRP), Nucleocapsid (N) and Ribonuclease P (RP) genes.
What is the difference between the Negative Extraction Control and Positive Extraction Control (RP) in the table “Expected Performance of Controls” in the datasheet? What reagents are added in these 2 wells?
The Negative Extraction Control (NEC) monitors the potential contamination from positive samples into negative samples during the nucleic acid extraction process. It is used to ensure that the sample is not contaminated during the assay. NEC is the kit component, K1461-100-4 and it contains Human Embryonic Lung cells. NEC should be extracted at the same time when the researcher extracts the RNA from the experimental patient samples.
The Positive Extraction Control (PEC) is the experimental patient sample. It is not a component of the kit. If the sample is positive for Ribonuclease P (RP) under the HEX channel, it shows that the RNA was successfully isolated from the patient sample. If the sample does not show a Ct value for RP, then the extraction process is not valid and the patient sample needs to be re-collected and re-extracted.
The Positive Extraction Control (PEC) is the experimental patient sample. It is not a component of the kit. If the sample is positive for Ribonuclease P (RP) under the HEX channel, it shows that the RNA was successfully isolated from the patient sample. If the sample does not show a Ct value for RP, then the extraction process is not valid and the patient sample needs to be re-collected and re-extracted.
The expected results and Ct values are the same for the Positive Extraction Control and Negative Extraction Control (negative, negative, positive results) in the table “Expected Performance of Controls”. Why are the results same for these two different wells?
The Positive Extraction Control and Negative Extraction Control are different samples but these two samples/wells will show the same results (negative N, negative RdRP, positive RP) to serve different purposes.
1. Negative Extraction Control provided in the kit includes human embryonic lung cells so it will always be positive for RP. It is called "Negative Extraction Control"(NEC) because it should not be positive for the viral genes. If NEC shows positive results for RdRP or N, it means that there was some contamination during the nucleic acid extraction process.
2. Positive Extraction Control (PEC) is an experimental sample. If the experimental sample is negative for COVID-19, then PEC should show negative results for N and RdRP and positive results for RP.
In summary, NEC is used to make sure that there is no contamination during the nucleic acid extraction process whereas PEC is used to make sure that that the nucleic acid extraction was done correctly for the samples.
1. Negative Extraction Control provided in the kit includes human embryonic lung cells so it will always be positive for RP. It is called "Negative Extraction Control"(NEC) because it should not be positive for the viral genes. If NEC shows positive results for RdRP or N, it means that there was some contamination during the nucleic acid extraction process.
2. Positive Extraction Control (PEC) is an experimental sample. If the experimental sample is negative for COVID-19, then PEC should show negative results for N and RdRP and positive results for RP.
In summary, NEC is used to make sure that there is no contamination during the nucleic acid extraction process whereas PEC is used to make sure that that the nucleic acid extraction was done correctly for the samples.
What is the Negative Extraction Control provided in the Kit?
The Negative Extraction Control provided in the Kit includes HEL (Human Embryonic Lung) cells. The Negative Extraction Control should be extracted and processed in the same way as samples are processed and should not be used directly in RT-qPCR reactions.
Is the Primer and Probe for the internal control RP provided in the Kit?
Yes, the Primer and Probe for the internal control RP are included in the COVID-19 Primers/Probes, K1461-100-1.
What is the bp size of the amplicon for the RNA-dependent RNA polymerase (RdRP), nucleocapsid (N) and RP targets?
The Coronavirus Rapid RT-qPCR Detection kit is specific for the RNA-dependent RNA polymerase (RdRP) and nucleocapsid (N). The amplicon sizes for these proteins and the internal control, RP are given below:
N2: 67 bp
RdRP: 108 bp
RP: 65 bp
N2: 67 bp
RdRP: 108 bp
RP: 65 bp
What is the Positive Control provided in the Kit?
The Positive Control is a DNA plasmid containing the sequences of RNA-dependent RNA polymerase (RdRP) and nucleocapsid (N). The Positive Control is provided to make sure that the kit components are all working.
