LifeDireX - QP019-0100

COVID-19 RT-qPCR Detection Kit (RUO)

Size: 100 Rxns



Note: COVID-19, SARS-CoV-2, RT-qPCR, RdRP, N
Categories: Real-Time PCR Reagent

In view of the joint global efforts of advancing collaborative research in diagnostics, therapeutics, and vaccination in the fight against the COVID-19 (SARS-CoV-2) pandemic, Bio-Helix has specifically developed the LifeDireX COVID-19 RT-qPCR Detection Kit for human respiratory tract specimens. The kit is characterized by: (1) High specificity for the RdRP and N target markers (2) Data obtained in less than 2 hours; and (3) Compatible with standard RT-qPCR machines (ABI 7500, Bio-Rad CFX96, QuantStudio’s 7 Flex). 

Part No.

Component

Volume

Reactions/Kit

 QP019-0100-1 

 2X RT-qPCR MasterMix

1.25 ml 

100 

 QP019-0100-2

 RT-qPCR Enzyme Mix

40 μl 

100 

 QP019-0100-3

 COVID-19 Primers/Probes

200 μl 

100 

 QP019-0100-4

 Positive Control

100 μl 

20 

 QP019-0100-5 

 Negative Extraction Control

1.0 ml 

20 

 QP019-0100-6

 Nuclease-Free Water

1.0 ml 

100 

Application

  • Gene Expression (mRNA) Analysis
  • Copy Number Analysis
  • SNP Genotype Analysis

Important notes

  1. Shake gently before use to avoid foaming and low-speed centrifugation.
  2. Reduce the exposure time.
  3. During operation, always wear a lab coat, disposable gloves, and protective equipment.

Q: Which genes does the Kit target?
A: It covers 3 genes: N, RdRP, and RP genes

Q: What are the shipment measurements?
A: We want to make sure our kits get to the destination at a cooling temperature, therefore we use Dry Ice and Ice packs in combination.

Q: Is the kit CE or FDA accredited?
A:  This kit is for Research Use only. 

Q: Can we run the kit in any equipment?
A: Our kit is compatible with RT-qPCR machine standards and our selected one is Bio-Rad CFX96. Please let us know which machine you are using. You are welcome to try the machine of your choice.

  1. PCR Reaction: Thaw and assemble the following components in a 0.2 ml PCR tube on ice just prior to use: COVID-19 Primers, COVID-19 Probes, 2X RT-qPCR MasterMix, and RT- qPCR Enzyme Mix. Caution: Do not add more than one RNA sample into a single qPCR tube. Mix gently. If necessary, centrifuge briefly.

 

Component 20 μl Patient Sample 20 μl Positive Control 20 μl Positive Control Negative Control
RNA Sample 5 μl 0 μl 0 μl 0 μl
COVID-19 Primers/Probes 2 μl 2 μl 2 μl 2 μl
2X RT-qPCR MasterMix 10 μl 10 μl 10 μl 10 μl
RT-qPCR Enzyme Mix 0.4 μl 0.4 μl 0.4 μl 0.4 μl
RT-qPCR Enzyme Mix 0 μl 5 μl 0 μl 0 μl
Negative Extraction Control 0 μl 0 μl 5 μl 0 μl
Nuclease – Free H2O 2.6 μl 2.6 μl 2.6 μl 7.6 μl
  1. Use the Nuclease-free H2O for the Negative Control while using Positive Control for the Positive Control setup. Cap tubes and place in the thermal cycler.
  2. Process in the thermal cycler for 42 cycles as follows:
Steps Temperature/Time Cycle
cDNA Synthesis 15 minutes at 42°C 1
Pre-Denaturation 10 minutes at 95°C 1
Denaturation 15 seconds at 95°C 40
Annealing 60 seconds at 60°C
Melting curve Refer to specific guidelines for instrument used  

Note: Optimal conditions for amplification will vary depending on the primers and thermal cycler used. It may be necessary to optimize the system for individual primers, template, and thermal cycler.

  1. Detection: As three channels (FAM, ROX, HEX) are used in this one tube qPCR assay, we recommend to perform the channel calibration as requested by its manufacturer. Please refer to the instrument’s user manual to perform this calibration. Choose the FAM, ROX, and HEX channels for each sample to be tested with the LifeDireX COVID-19 RT-qPCR Detection Kit. Select “None” for ROX passive reference on any qPCR machine requiring ROX as the reference dye.

 

Expected Performance of Controls

Control Type Used to Monitor Expected Results and Ct Values
N (FAM) RP (HEX) RdRP (ROX)
Positive Flawed assay setup and reagent failure, including degraded primer and probe

Positive Ct < 40.0

Negative Ct ND

Positive Ct < 40.0

Negative

(“NTC”)

Assay or extraction reagent

contamination

Negative

Ct ND

Negative

Ct ND

Negative

Ct ND

Negative

Extraction Control

Cross-contamination

Negative

Ct ND

Positive

Ct < 40.0

Negative

Ct ND

ND = Not Dtected.

Results are considered invalid if any control does not perform as specified above.

 

Interpretation of Results

SARS-CoV-2 Interpretation
N RdRP RP
+ + +/- Positive
If only one of the two targets is positive +/- Inconclusive Result
  - + Negative
- - - Negative

Note: The results from this kit should be interpreted in combination with all relevant laboratory findings.


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