The polymerase chain reaction (PCR) is one of the fundamental technologies in genetics. It enables the
identification and up-copying of DNA sequences so that they can be examined and checked for genetic
defects. Novogenia’s laboratory, founded in 2009, is approved by the Austrian Federal Ministry of
Health to perform medical genetic analyses.

Analyses per month

To ensure the required quality, a test for SARS-CoV-2 involves three individual PCR analyses. With our
existing laboratory equipment, we can, therefore, smoothly perform 5,000 tests for SARS-CoV-2 per
day.

The technical equipment of our laboratory

Extraction of the viral RNA

In the first step, the viral RNA must be extracted from the throat swab. This process can be performed
both manually and automatically. Since manual extraction can only process 30 samples in one working
hour, Novogenia’s extraction process is fully automated.

For this purpose, we have six “Qiagen Biorobot Universal” extraction machines, four of which are
currently reserved for the extraction of viral RNA.

Each biorobot can extract 192 samples in 90 minutes and thus has a capacity for around 3,000
extractions per day so that we can currently process around Twelve thousand incoming samples every
day. If necessary, we can increase our capabilities at short notice at any time.

Used analysis plate formats

Many laboratories use 96-well plates for their analyses. With three individual analyses per sample,
approximately 30 analyses for SARS-CoV-2 can be performed on one plate. Novogenia, on the other
hand, uses 384-well plates so that about 125 analyses can be performed on one plate.

The plates loaded with samples are loaded into the analyzer by automatic loading robots. One analysis
station can hold up to 100 plates with around 12,400 individual samples, which can then be processed
without human intervention.

Analysis of the virus genes

The analysis of the viral RNA is performed with a so-called real-time PCR cycler. Novogenia uses up to
seven such devices in total. For our SARS-CoV- 2 PCR analyses, we use two instruments of the model
Viia 7 from Thermo Fisher.

The analysis takes 90 minutes per plate. With three samples per test person, a PCR cycler performs
around 3,000 fully automated tests for SARS-CoV-2 per day, with a total of 9,000 individual PCR
analyses.

If necessary, we can convert our other PCR cyclers at short notice and increase the capacity for testing
for SARS-CoV-2 from currently 6,000 to well over 10,000.

The validation of the analytical system

Before an analysis system is put into clinical use, it must first be validated in several steps.

Reagents for clinical diagnostics with CE-IVD certification

Although not mandatory, reagents with CE-IVD certification are preferable. These have been validated in advance by the manufacturer, registered as medical products, and subjected to various quality tests.

Novogenia uses reagents (PhoenixDX SARS-CoV-2 IVD) from the ISO certified company Procomcure from Thalgau, Austria. The company specializes in the development of reagents with CE-IVD certification and, as a domestic producer, is hardly affected by the current problems with international supply chains.

Internal quality controls for reliable test results

The widely used analytical protocol of WHO and Charité is considered as an emergency guide for
laboratories to increase analytical capacities. Although it works very well in principle, in our opinion
the system is missing an important point. This is because the protocol does not provide for quality
control of the samples to be analysed so that theoretically, even an empty throat swab could be
examined, which would inevitably result in a negative test result.

In this respect, the Procomcure reagents we use have the advantage that they analyse human genes
as well as viral genes. If the analysis for human RNA works, the stability of the sample is ensured, and
the result of the test for SARS-CoV-2 is therefore reliable.

The analytical accuracy of the reagents used

To assess the accuracy of a systematic system, it is necessary to know the sensitivity and specificity of
the system. Sensitivity is the ability to detect positive samples, while specificity indicates how reliably
negative samples are detected. A rate of 100 percent means that all positive or negative samples have
been correctly identified.

As part of the CE-IVD certification process, 53 clinical samples from subjects with COVID-19 symptoms
were analysed with Procomcure’s test kit and compared to the results of an already validated
reference test kit.

The following values were determined:

Do the reagents mistakenly detect other viruses?

To check whether the reagents react erroneously to other viruses, they were tested against 60
different pathogens. The analysis system only detected samples with SARS-CoV-2.

Virus load required for a positive test result

During the peak of infection, millions of viruses are detectable in the throat and, therefore, easy to
analyse. However, the viral load is usually very low in the early stages of the infection. It is, therefore,
important that an analysis system can detect the smallest possible number of viruses in the sample
under investigation. To measure the analytical accuracy, viral RNA was continuously diluted and then
analysed 24 times with the system.

With only 50 viruses in the sample, they could be identified with an accuracy of 95%. For a single virus
in the sample, the accuracy was still 25%. The system is, therefore, able to detect very few viruses in a
sample and to detect the infection at a very early stage.

Validation in the Novogenia laboratory

To measure sensitivity and specificity in Novogenia’s laboratory, an internal validation was performed. For this purpose, 113 samples were used:

Positive confirmed samples: 12

negative samples: 97

Positive confirmed samples were verified by Roche’s CE-IVD certified COBAS system (SARS-CoV-2).

With a sensitivity of 100% and a specificity of 100%, Novogenia’s laboratory thus provides a very high analytical accuracy.

Further validation in progress

Further validation is currently in progress. This will proceed as follows:

  • 200 samples consisting of clinically positive and clinically negative samples
  • Result confirmed by CE-IVD ROCHE COBAS System
  • Result confirmed by PhoenixDX SARS-CoV-2 IVD test kit (performed by Procomcure)
  • Result confirmed by PhoenixDX SARS-CoV-2 IVD test kit (performed by Novogenia)
  • Result confirmed by the Thermo Fisher SARS-CoV-2 IVD test kit (shown by Novogenia)

This validation will provide robust information on the sensitivity and specificity of all three analytical
systems:

  • Roche Cobas CI-IVD
  • PhoenixDX SARS-CoV-2 IVD
  • Thermofisher SARS-CoV-2 IVD

This means that we then have validated alternative systems at our disposal to verify unclear results or
to switch to other reagents in the event of a supply bottleneck.

The sample sets from Novogenia

In addition to mass analyses, for which institutions can collect the samples, Novogenia can also carry out the analysis of samples from individuals. For this purpose, we have developed sample sets that kill viruses and can also preserve a low virus load at room temperature for at least seven days.

A mailing of the sample kit to the patient and a return to the laboratory by mail is therefore conceivable. The sample sets are currently undergoing CE-IVD certification.

Stability of viral RNA at room temperature

The graph shows the positive detection (yellow line) of a throat swab sample after it has been taken
by the patient herself and stored at room temperature for seven days before extraction.

Authorization for SARS-CoV-2 analyses

Novogenia is registered as an analytical laboratory for SARS-CoV-2 by the Federal Ministry of Health and integrated into the notification system for notifiable disease.

With a machine capacity of 10,000 analyses per day and a reagent supplier who can supply us with the
necessary quantities, Novogenia can carry out a large part of the 15,000 analyses required daily in
Austria.