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Novel Coronavirus (2019-nCoV) Nucleic Acid Detection Kit (Fluorescent RT-PCR)

The kit is a TaqMan probe real-time fluorescent PCR to qualitatively detect ORF1ab, N genes nucleic acid of 2019-nCoV in throat swab, sputum in patients with suspected pneumonia symptoms and Clustering cases.

  • Item NO.:

    NRM-FT-400-48
  • Payment:

    T/T
  • Specification:

    48 Tests
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  • Product Detail

【Intended Use】

The kit is a TaqMan probe real-time fluorescent PCR to qualitatively detect ORF1ab, N genes nucleic acid of 2019-nCoV in throat swab, sputum in patients with suspected pneumonia symptoms and Clustering cases.

 

The test results are for clinical reference only and cannot be regarded as only benchmark for clinical diagnosis.it’s recommended to make manifested analysis with regarding to clinical symptoms and other laboratory results.

 

Operators should have experience of professional training in gene amplification or molecular biology method testing, and have relevant qualifications for laboratory operations. The laboratory should have reasonable biological safety precautions and protective procedures.

 

Test Principle

In this kit, primers and probes are designed to the conserved and specific regions of the ORF1ab, N of 2019-nCoV, respectively. Fluorescent PCR is used to detect viral RNA after nucleic acid purification of sample RNA.

 

This test kit uses real-time fluorescent PCR technology.During PCR amplification, the probe binds to the template, and the 5'-end reporter group of the probe is cleaved by the Taq enzyme (5'→3' exonuclease activity), thereby moving away from the quenching group to generate a fluorescent signal. The real-time amplification curve is automatically plotted based on the detected fluorescence signal, and the sample Ct value is calculated(The number of cycles experienced when the fluorescence signal in each reaction tube reaches the settled threshold). FAM ,VIC and Texas red fluorophores are utalized to mark ORF1ab gene , N gene probes in reaction solution. By using one test, qualitative detection of the above two genes of 2019-nCoV can be performed simultaneously.

To avoid the false negative result, the kit is provided with an internal control (IC), which is marked with Cy5 fluorescent group which could be used to monitoring the extraction procedure and reaction solution.

 

Main Components

No.

Components

Ingredients

Specification

1

Nuclease-free

water

DEPC Water

1000μL ×1Tube

2

Nucleic acid

amplification

reaction solution

Tris-HClKClMg2+

528μL ×1Tube

3

Reverse

transcriptase

Reverse transcriptase, RNA enzyme inhibitor

48μL ×1Tube

4

O/N reaction

solution

ORF1ab, N genesand internal control primers and probes

288μL ×1Tube

5

Internal control

Human gene fragment

40μL ×1Tube

6

Positive control

(O/N)

Recombinant vector containing ORF1ab, N gene fragments

400μL ×1Tube

7

Negative control

Physiological saline

400μL ×1Tube

8

DNA Polymerase

DNA Polymerase

50μL ×1Tube

 

Note1: The different lots of components cannot be exchanged.

Note2:The test kit doesn’t contain nucleic acid extraction reagent,please prepare the reagent before use.

 

1. Sample handling

Add 10μL of internal control into 200μL of positive control and negative control, respectively in as to perform the nucleic acid extraction.

2. Reagent preparation:

System preparation: Take pre-installed Nucleic acid amplification lyophilizer,ensued by opening the foil pouch and centrifuge it immediately. Sample amount of each test should be the multiples of 8 containing one positive control and one negative control.add 35μL nuclease-free water onto each reaction tube.

3.Sample Loading:

Add 5μL Sample nucleic acid to be tested,positive control,negative control that the whole reaction volume is 40 μL in the allocated reaction solution.for it doesn’t necessitate a fine mixture,and just close the lid of  PCR reaction tube,centrifuge the liquid in the tube wall upon the bottom of tube,thus process the PCR amplification.

After verification,it’s estimated that addition of 5μL-10μL Paraffin oil in the PCR system will not affect the PCR result but avoid contamination and liquid vaporization.

 

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