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COVID-19 Nucleic Acid Detection Kit (RT-NC-qPCR Fluorescence probe method)

COVID-19 Nucleic Acid Detection Kit (RT-NC-qPCR Fluorescence probe method)

Note: Price/availability/specifications subject to change without notice. Unless otherwise indicated, our catalog and customized products are for research use only and not intended for human or animal diagnostic or therapeutic use.

Product Description

[Application]

It can be used for qualitative and quantitative detection of COVID-19 RNA in throat swabs, bronchoalveolar lavage fluid and plasma samples, and also can be used for screening and auxiliary diagnosis of Novel Virus Infection.

[Principle] ··

Two sets of primers and fluorescent probes were designed to cover simultaneously the POL gene region of COVID-19 strain named AY27874, and the internal reference gene was designed. RT-NC-qPCR technique was used for real-time quantitative monitoring of COVID-19 nucleic acid. 

[Kit Components] 

Description

Quantity

Storage condition

Lysis solution

50 ml

4°C

RT-PCR Reaction Mixture (RT-PCR)

100 reactions/package

4°C

Primer probe mixture (P-M)

100 reactions/package

4°C

Positive control

50 μl

4°C

[RNA extraction: one of the following]

1. To Throat Swab sample: 100 μl lysis solution was prepared in a 1.5 ml test tube. The head of the throat swab was immersed in the 1.5 ml test tube for 2-3 seconds, and the throat swab was discarded in 75% ethanol solution. Heated the sample at 60°C for 5 minutes, then at 90°C for 10 minutes to obtain RNA solution.

2. To plasma sample: 20 μl plasma was mixed with 100 μl lysis solution in a 1.5 ml test tube. The test tube was then heated at 60°C for 5 minutes, then heated at 90°C for 10 minutes. 

3. To bronchoalveolar lavage fluid: Mix 50 μl bronchoalveolar lavage fluid with 100 μl lysis solution in 1.5 ml test tube, followed by incubation at 60°C for 5 minutes, at 90°C for 10 minutes. 

[PCR Reaction]

Component 

Volume for one reaction

RT-PCR Reaction Mixture?RT-PCR?

5 μl

Primers probe(P-M)

10 μl

RNA 

5 μl

[RT-NC-qPCR Thermal Cycling Parameters]

Step

Stage

No. of cycles

Temperature

Time

RT

1

1

42°C

10 minutes

Enzyme inactivation

2

1

95°C

3 minutes

PCR Amplification

3

40

95°C

10 seconds

46°C

30 seconds

60°C

30 seconds

FAM and TET fluorescence signals were detected at 60°C. FAM corresponds to novel coronavirus-specific ORF1ab and N Genes, TET Corresponds to human ACE2 gene.

[Quality Controls] 

1.Negative control: 5 μl lysis solution as the template. 

2. Positive Control: 5 μl Positive Control Solution as the template. 

[Result interpretation]

1. Positive and negative controls: The test is valid under the condition that positive control has Ct reading while negative control has no Ct reading.

2. If negative control has Ct reading, indicating cross contamination occurred, the test is not reliable and need to repeat.

3. If the positive control is negative, indicating the PCR amplification is failed, therefore the test is invalid and needs to repeat. 

4. Negative results: The amplification curve of TET fluorescence has Ct value, and that of FAM fluorescence has no Ct value.

5.Positive results: Both FAM and TET fluorescence amplification curve have Ct value.

6. Invalid sample: both FAM and TET fluorescence amplification curve have no Ct value indicating the poor quality of sample.

 

This product has obtained CE certification.

 

 

[Contact]

Aicon Biotech,Inc
3800 SW Cedar Hills Boulevard, Suite 227, Beaverton, OR,97005

 

Features

  • 999 Units in Stock

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Shipping info

$50.00 to United States

Contact us to order

+1(206)6369998 +1(503)2681752


This product was added to our catalog on Tuesday 17 March, 2020.