SARS-CoV-2(2019-nCOV) Coronavirus and Influenza A/B Virus Multiplex RT-qPCR Detection Kits,ShineGene_specification/price/image

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SARS-CoV-2(2019-nCOV) Coronavirus and Influenza A/B Virus Multiplex RT-qPCR Detection Kits

Catalog #	Pack size	Price($)
COV00102	50tests	1200.00
COV00103	100tests	2160.00

This kit is used for in vitro diagnostics for 2019-Novel Coronavirus (2019-nCoV) infection including suspected case, suspected aggregative diseases and other scenarios. By testing the suspected person’s respiratory specimen, serum or blood, the kit can tell if the specimens contain any virus RNA, so it can be used for clinical screening of suspected cases.
       Select the 2019-nCoV N gene and E gene as the target amplifying area, design specific primer and fluorescent probes, use FAM label, detect if the specimen contains any 2019 nCoV RNA. Select the 2019-nCoV ORF1ab gene, design specific primer and florescent probes, use ROX label, detect if the specimen contains any 2019-nCoV RNA. Select influenza A/B specific sequence, design specific primer and fluorescent probes, use Cy5 label, detect if the specimen contains any influenza A/B virus RNA. Select other species’ sequence (none-human and none-targeted sequence) as internal primer/probe, label with VIC, monitor the whole process.
        The kit also contains UNG enzyme anti-contamination system. Its theory is it selectively hydrolyzes Uracil glycoside bond in the single or double chain of DNA that contains dU. This leads to a DNA chain with nucleotide deletion which will further hydrolyzed in alkaline media under high temperature and thus avoid contamination by PCR amplification.

                                                                        4 Fluorescent Signals and Target

	Fluorescent signal	Target
1	FAM	2019-nCoV N gene and E gene
2	ROX	2019-nCoV ORF1ab gene
3	Cy5	Influenza A/B specific sequence
4	VIC	MS2 internal standard (RNA Phage pseudo virus)

Key Features:
1.High sensitivity 500 copy/ml
2.Very specific, can detect 2019-nCoV E gene, N gene and ORF1ab different zone sequence at the same time in one tube.
3.Can simultaneously identify and distinguish influenza viruses to avoid misdiagnosis.
4.One-step RT-PCR, strong amplification signal, high stability.
Storage condition: -20°C.
Equipment Requirements: Fluorescent PCR amplifier; Fluorescent PCR amplifier with 4 colors or multi-color fluorescent PCR amplifier such as ABI Prism®7500, Bio-rad CFX96 etc.
Set up reaction mixture:

Component	Volume（μL）
RT-mix	Nx 9
Enzyme mix	Nx 1.5
2019-nCoV primer probe	Nx 1
Influenza A/B primer probe	Nx 1
Total volume	Nx 12.5

Set up cycle program::

Step		Temp	Time	Cycles
1	UNG Incubation	25°C	2min	1
2	RT incubation	50℃	10min	1
3	Enzyme activation	95℃	2min	1
4	Denature	95℃	5sec	45
4	Annealing, extension, detction*	60℃	35sec	45
*Fluorescent test in extension: Channel for 2019-nCoV E gene and N gene is FAM; Channel for 2019-nCoV ORF1ab gene is ROX; Channel for influenza A/B is Cy5; Channel for internal standard is VIC;

Data Analysis:

Ct Value*	Result	Recommendations
Ct（FAM）≤36.5 and Ct（ROX）≤38	2019-nCoV positive(+)	Active treatment
Ct（FAM、ROX）≥39 or no value，and Ct（VIC）＜40	2019-nCoV negative(-)	More Rest
Ct（FAM、ROX）≥39 or no value，and Ct（VIC）＜40,but Ct(Cy5) ≤36.5	2019-nCoV negative(-) Influenza A/B positive(+)	Active treatment
Ct（FAM）≤36.5 and Ct（ROX）≤38,and Ct(Cy5) ≤36.5	2019-nCoV negative(+) Influenza A/B positive(+)	Active treatment
only Ct（FAM）≤36.5 or Ct（ROX）≤38	Inconclusive Result	Repeat extraction and double confirm. After getting Ct（FAM）≤36.5 or Ct（ROX）≤38, the result is positive
36.5＜Ct（FAM）＜39 or 38＜Ct（ROX）＜39	Inconclusive Result	Increase specimen concentration and double confirm. If Ct（FAM）＜39 and Ct（ROX）＜39, it means 2019-nCoV result is positive; other results need be analyzed by other methods.
Ct（FAM、ROX、CY5）≥39 or no value，and Ct（VIC）≥40 or no value	Invalid Result	Indicates that there might be issues with experiment itself, reagent, need check before repeat the experiment

SARS-CoV-2(2019-nCOV) Coronavirus and Influenza A/B Virus Multiplex RT-qPCR Detection Kits
Handbook of COVID-19 Prevention and Treatment
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