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 is versatile, can be run on a variety of different equipments with a running time of ~1 hour (minus the RNA extraction time) and up to 95 specimens per run(depending on the Real Time PCR machine used), 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 lower respiratory specimens such as sputum.
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. Fluorescence intensity is monitored at each PCR cycle in corresponding channels at a threshold of 0.05.
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 (>95% sensitivity and specificity in detection of SARS-COV-2, great linearity (Correlation coefficient (R2) between threshold cycle (Ct) values and quantification values (log10) in each tested concentration ranged between 0.9929~0.9999), great sensitivity (LOD of 10 copies) and specificity (as proven by in silico analysis and wet testing) 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. Refer to our Performance evaluation documents for information about our results, which were proven on multiple clinical specimens and FDA approved BEI materials.
This product contains Taq polymerase, reverse transcriptase, dNTPs, 2x reaction buffer, Hot Taq, 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.