The GoodGene COVID-19 QPlex RT-PCR Kit was designed to detect open reading frame (ORF1ab), RdRP, and nucleoprotein (N) based on the genomic database of COVID-19. Reverse transcription and RT-PCR are carried out sequentially in the same tube.
GG COVID-19 Quadplex real time RT-PCR Assay includes primers and dual-labeled hydrolysis (Taqman®) probes to be used in the in vitro qualitative detection of COVID-19 virus RNA isolated from clinical specimens including nasopharyngeal, oropharyngeal, and nasal swab.
For the RdRP specific probe, the signal from the fluorescent dye (FAM) on the 5’ end is quenched by BHQ-1 on its 3’ end. For the N specific probe, the signal from the fluorescent dye (Texas Red) on the 5’ end is quenched by BHQ-2 on its 3’ end. For the Orf1ab specific probe, the signal from the fluorescent dye (Cy5) on the 5’ end is quenched by BHQ-1 on its 3’ end. For the internal control beta actin specific probe, the signal from the fluorescent dye (Cy5) on the 5’end is quenched by BHQ-1 on its 3’ end.
The real-time PCR instrument detects this fluorescence from the unquenched dye. With each cycle of PCR, more probes are cleaved resulting in an increase in fluorescence. Fluorescence intensity is monitored at each PCR cycle in corresponding channels at a threshold of 0.05.
Kit constituents
| Constituents | Purchase option #1 | Purchase option #2 | Purchase option #3 |
|---|---|---|---|
| (1) Primer/probes | o | o | o |
| (2) Postive (ORF and N gene of COVID-19) and internal controls | o | o | o |
| (3) Reverse transcription (RT)-PCR premix (one step reverse transciptase/Taq polymerase enzyme, RT-PCR buffer, dNTP, RNase free water) | X | o | o |
| (4) RNA extraction kit | X | x | o |
| Pros | Fastest* | Cheapest | May be convenient |
| Constituents | Less convenient if short on supply** | Fast and cheap | Slowest |
*small in volume, enables fast dhl/fedex shipping from South Korea.
**Have to buy from other distributors and may have to validate other components of testing.
With option #2, all components required for reaction are added during setup, and there is no need to add additional components once the reaction has been started, which is a very fast and will give satisfactory results in most cases. The kit showed excellent performance (100% sensitivity and specificity in detection of SARS-COV-2 and LOD of 5 to 100 copies) in clinical and performance evaluation carried out according to FDA EUA guidelines with clinical specimens and materials from BEI Resources (Manassas, VA, USA) and more.
This product contains polymerase, reverse transcriptase, dNTPs, 2x reaction buffer, RNase inhibitor, specific primers and probes for detection except template RNA (from patient specimen) and RNA extraction kit. You just need to add distilled water and template RNA for RT-PCR and the RT-PCR is done in a single tube.
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