Detecting COVID-19 and influenza simultaneously with a single sample

Polymerase chain reaction (PCR) is considered an essential technique deployed in medical and biological research labs for numerous applications.

Detecting COVID-19 and influenza simultaneously with a single sample

Image Credit: Lumex Instruments

Applications include qualitative and quantitative nucleic acid analysis, the diagnosis of hereditary diseases, the detection and diagnosis of infectious diseases, DNA cloning for sequencing, DNA-based phylogeny, functional analysis of genes, and the identification of genetic fingerprints (used in forensic sciences and paternity testing).

AriaDNA real-time PCR analyzer provides highly specific and sensitive qualitative and quantitative nucleic acids analysis in various samples. Microchip real-time PCR technique with the extremely fast heating and cooling device performs rapid amplification and analysis in micro-volumes.

Features of microchip PCR analyzer «AriaDNA»

  • Short PCR-runtime due to high thermocycling rate
  • Low detection limit
  • Low sample and reagents consumption
  • Simultaneous qualitative and quantitative DNA/RNA analysis
  • Flexibility and customization
  • Small footprint and low energy consumption
  • Dedicated software complies with FDA 21 CFR part 11

A microchip is a thin plate made of heat-conducting material with tiny wells — microreactors. There are two formats of microchips:

Preloaded microchips with lyophilized reagents (ready-to-use) are designed for specific applications in the areas of medicine, veterinary, and agriculture.

Empty microchips can be used by the end-user for a number of customized applications with any compatible third-party PCR kits.

Application areas

1. Veterinary

  • Avian Influenza virus and pathogens
  • Cattle diseases pathogens
  • African swine fever
  • Fish disease pathogens

2. Agriculture, Food safety

  • Grapevine diseases
  • Potato bacterial and viral pathogens
  • Pathogens in raw and processed food
  • Genetically modified lines (maize, oilseed rape)

3. Clinical diagnostics, Healthcare

  • COVID-19 detection system, incl. detection from saliva samples
  • Influenza A, Influenza B
  • SNPs in human genome (Thrombophilia, Warfarin)
  • Sexually transmitted infections (STI)

and many more.

Influenza caused by influenza viruses and COVID-19 caused by the SARS-COV-2 virus are both respiratory tract infections that exhibit similar symptoms.

Detecting COVID-19 and influenza simultaneously with a single sample

Image Credit: Lumex Instruments

Moreover, unique strains of influenza A and B viruses such as H1N1, H2N2, H3N2, H1N1, and H1N1/09 subtypes, respectively, can be unpredictable and may lead to pandemics such as those in 1918, 1957, 1968, 1977, and 2009.

In the event of coinfections of these viruses, particularly during the current COVID-19 pandemic, it is crucial to differentiate and identify the precise causative virus for suitable therapeutic intervention.

While current molecular-based tests provide a reliable and accurate solution to differentiate between influenza and SARS-CoV-2 virus from the same sample, a rapid, affordable, and less labor-intensive technique is still required for the optimization of clinical strategies and societal benefits during pandemic bottlenecks.

Lumex Instruments Canada has created an accurate, cost-effective, and fast Microchip RT-PCR Influenza and COVID-19 detection method from upper respiratory tract samples (swabs).

The test kit utilizes a pre-loaded microchip kit with the primers and probes lyophilized in the microchip for testing of SARS-CoV-2 and the Influenza A & B viruses. It is a research use only (RUO) application.

Detecting COVID-19 and influenza simultaneously with a single sample

Image Credit: Lumex Instruments

The N1 primers-probe target the nucleocapsid gene (N) gene of SARS-CoV-2 virus, while the Inf-A primers-probe are targeted to focus on matrix (M1) gene of Influenza A virus and Inf-B primers-probe target nonstructural 2 (NS2) gene of Influenza B virus.

The HsRPP30 primers-probe is utilized as an internal control that targets the RNase P gene that appears in the human genome. The US Center for Disease Control and Prevention (CDC) has created RT-PCR assays and issued directives for the appropriate detection of these viruses.

Features and benefits:

  • Existing real-time PCR assays and PCR instruments use large volumes of reagents (20 μl reaction). The test can be expensive and can be burdened by the bottleneck of PCR reagent supply in the event of disease outbreaks.
  • Major benefits of the Microchip RT-PCR Influenza and COVID-19 detection system include limited reagent consumption (1.2 μl reaction), rapid analysis, transportation under ambient conditions, and handy processing of PCR analysis.
  • Ease-of-use with lyophilized PCR reagents in the microchips will boost the reliability of laboratory analysis considerably. It will also improve fast-responder settings by significantly limiting any operator-associated errors.
  • The Microchip RT-PCR Influenza and COVID-19 detection system – compact and low-energy requiring – is ready to be utilized as a point-of-analysis network.
  • Early detection of pathogens will reduce the economic loss caused by outbreaks. This technology would boost public health maintenance and enhance the validity of quarantine measures. Microchip kits can be developed for specific pathogen applications in future outbreaks.

User-friendly software

The software is developed for real-time PCR data acquisition while facilitating simplified operation steps. It provides auto-interpretation of results, enables manual analysis of data, and prints a report in accordance with 21 CFR part 11 requirements.

Fluorescence detection

Two detection channels of the AriaDNA analyzer record the following panels of targets:

Source: Lumex Instruments

. ..
Channel 1 (FAM) 1. Influenza A, and 2. SARS-CoV-2 N-gene N1
Channel 2 (Cy5) 1. Influenza B, and 2. Human RNase P gene
HsRPP30 (Internal Control)

 

Layout of microchip

Detecting COVID-19 and influenza simultaneously with a single sample

Image Credit: Lumex Instruments

Test panel:

  • Inf-A (Influenza A)
  • Inf-B (Influenza B)
  • nCoVN1 (SARS-CoV-2 N1)
  • HsRPP30 (Internal Control)

Controls:

  • NEC (Negative extraction control) - run with NEC sample
  • PTC (Positive template control) - run with Influenza A & B RNAs, and SARS-CoV-2 RNA • NTC (No template control) - run with nuclease-free water

A number of samples per chip:

  • 12 patient samples

Analysis flow chart

  1. Pre-loaded microchips: Demand individual purchase of the following kits to run the test:
    1. RNA extraction kit: for the extraction of viral RNA from upper respiratory tract samples (swabs), use Norgen Biotek Total RNA Purification Kit. https://norgenbiotek.com/product/total-rna-purification-kit
    2. Master Mix solution: The master mix Quantabio UltraPlex 1-Step ToughMix is advised. Reduced consumption of the master mix per sample up to 3.3 times contrasted with the recommended volumes for traditional multiplex PCR.
  2. Test procedure: Mix the RNA extracted from the sample with the necessary reagents and then pour this mixture into the microchip by following the instruction manual (supplied with the microchips). Then place the microchip into the AriaDNA analyzer and conduct the analysis with a preset protocol on a computer.
  3. Estimated sample throughput: 7 microchips per day per instrument (8h work shift), i.e., 84 samples per work shift. A reasonable number of microchips to order per instrument per month is 250 (10 boxes). Increased numbers of microchips can also be blanket ordered.

Results

Acquire the RT-PCR results in real-time and print a report in 52 minutes.

The detection limit is equivalent to 1.5×103 copies for Influenza A, 1.5×103 copies for Influenza B, and 1.5×103 copies for SARS-COV-2 in 1 mL of the sample.

About Lumex Instruments

For almost 30 years, Lumex Instruments has been developing and manufacturing laboratory and industrial analytical equipment, instruments, and procedures.

Our products are used for environmental control, water and wastewater treatment, agriculture, food and beverage quality and safety control, energy industry, and pharmaceutical and biochemical applications. Lumex Instruments offers more than 120 applications for the measurement of organic and inorganic chemicals in liquid, solid, and gas.

We offer both traditional and new ingenious solutions, unique technologies and classical equipment, well-established reliable applications, and innovative methods. From the beginning of the company, we have developed, manufactured, and sold more than 16,500 instruments.

Our products include:

  • capillary electrophoresis systems
  • PCR analyzers
  • atomic absorption spectrometers (including mercury analyzers and monitors)
  • FTIR and NIR spectrometers
  • fluorimeters and more

We combine high-quality standards with a flexible customized approach because we care about the efficiency of our customer operations. All our innovations are targeted to make the analyses cost-efficient, easy to run, and the instruments easy to maintain and service. We proudly deliver our products and services all around the world and we enjoy working with our valued customers and partners in many industries and countries.

With customers in more than 90 countries worldwide, Lumex Instruments has several offices and multiple distributors around the world.

A significant number of our applications are developed in accordance with international standards, such as ASTM, ISO, EN, and EPA.

Facts and figures:

  • More than 30 models of analytical instruments developed starting from 1991
  • Over 20 patents
  • 20% market share of the portable mercury analyzers
  • Over 120 applications
  • Over 350 employees and 25% of them are involved in the development of new applications and instruments

The customers who selected Lumex Instruments include BP, Enel, Royal Dutch Shell, PETRONAS, PetroChina, Chevron, UNIDO, SGS, Carlsberg Group, Unilever, PepsiCo, Nestle, Princeton University, and others.


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Last updated: Sep 8, 2021 at 12:23 PM

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