COVID19 High Throughput Real Time RT-PCR Nucleic Acid Detection Kit

Details for Product No. ABIN6952765,
Target
Reactivity
Human, SARS Coronavirus-2 (SARS-CoV-2)
Application
Real Time PCR (rtPCR)
Options
Purpose This kit is used for the qualitative in vitro detection of novel coronavirus (SARS-CoV-2) nucleic acid in patient sera and respiratory specimens (including pharyngeal swabs and alveolar lavage fluid samples).
Analytical Method Qualitative
Cross-Reactivity (Details) No cross reactivity was identified when the primers and probes in this kit were evaluated against MERS, SARS H1N1 (new influenza A H1N1 influenza virus (2009), seasonal H1N1 influenza virus) , H7N9 virus, influenza B virus, enterovirus A, B, C, D groups, Legionella, Pertussis, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Klebsiella pneumoniae, Mycobacterium tuberculosis, Candida albicans, Candida lucida, Cryptococcus neoformans ,and the human genome
Characteristics The Coronavirus (SARS-CoV-2) High Throughput Real Time RT-PCR Nucleic Acid Detection Kit is based on the PCR method which uses a fluorescent probe and a specific primer to detect two specific regions within the novel coronavirus (SARS-CoV-2) RNA genome. One target corresponds to the nucleocapsid protein N gene region, and the second target corresponds to the ORF1ab region. This molecular panel aids in the detection of viral RNA from SARS-CoV-2, the causative agent of COVID-19.

Primer-probe sets for the N and ORF1ab detect SARS-CoV-2 specifically, with no expected false positives from other coronaviruses or human microflora. The kit also includes a primer-probe set specific to the to the human housekeeping gene, ribonuclease P (RNP), are included to serve as an internal reference to monitor sample collection, RNA extraction, and amplificatio n.

This kit combines the 3 primer-probe sets into a single reaction, reducing the number of wells and reagents needed by as much as 75 % compared to other RT-PCR assays. The N and ORF1ab primer-probe sets are both detected using a 494/518nm (FITC or FAM) channel, and the RP gene is detected using 533/544nm channel.
Components
  • PCR Reaction Solution (Buffer, dNTP's, Primers, and Probes) 1300 μL
  • PCR Enzyme Mix (Reverse transcriptase (RT) enzyme, DNA polymerase) 96 μL
  • Positive Control (Stabilized RNA for the N gene and plasmid DNA for the RP gene) 4 x 100 μL
  • Negative Control (Nuclease-free water) 100 μL
Material not included
  • Fluorescence qPCR instrument capable of reading FAM and JOE or equivalent channels simultaneously (494 nm maximum absorption, 518 nm maximum emission, 533 nm maximum absorption, 544 maximum emission, respectively).
  • Vortex Mixer
  • Microcentrifuge
  • Pipettes
  • Sterile nuclease-free pipette tips and microfuge tubes
  • RNA extraction kit (e.g., Qiagen RNeasy® Mini Kit, EZI DSP Virus Kit, or similar)
Target Name (Antigen)
Alternative Name SARS-Coronavirus-2
Application Notes Limit of Detection: the minimum detection limit of this kit is 4 copies / μL in the reaction (16 copies / uL in the purified extract).
Repeatability: Precision testing showed that the coefficient of variation of the precision Ct values within this kit lot are ≤ 5 % .
Accuracy: The kit was tested with a third-party sourced positive reference product and the compliance rate was 100 %. The kit was tested with a third-party sourced negative reference product and the compliance rate was 100 %.
Comment

Amplification was not affected by: 20 μg/mL purified mucin, 100 μg/mL oxymetazoline hydrochloride, 125 μg/mL phenylephrine, 60 μg/mL sodium chloride, 50 μg/mL beclomethasone. 50 μg/mL dexamethasone, 100 μg/mL flunisolide, 100 μg/mL triamcinolone acetonide, 320 μg/mL budesonide, 100 μg/mL mometasone, 200 μg/mL fluticasone, 200 μg/mL of histamine dihydrochloride, 300 U/mL of [alpha] - interferon, 100 μg/mL zanamivir, 100 μg/mL of ribavirin, 100 μg/mL oseltamivir, 100 μg/mL peramivir, 100 μg/mL lopinavir, 100 μg/mL ritonavir, 100 μg/mL Abidor, 100 μg/mL azithromycin, 100 μg/mL tobramycin.

Restrictions For Research Use only
Storage -20 °C
Storage Comment The kit can be stored at -20°C for a period of 12 months prior to opening. After opening, the reagents are valid for 6 months if stored at -20°C. Up to 3 freeze-thaw cycles are permitted while retaining activity.