The symptoms of SARS-CoV-2 infection are fever, coughing, breathing difficulties and fatigue. Most patients suffer from a mild febrile illness with irregular lung infiltrates. Some patients, particularly elderly or chronically ill persons, develop severe acute respiratory distress syndrome (ARDS), which is fatal in three percent of cases. In February 2020, the disease caused by SARS-CoV-2 was named COVID-19 (coronavirus 19) by the WHO.

Laboratory tests for the diagnosis of SARS-CoV-2 infection include polymerase chain reaction (RT-PCR) with smears from the upper and lower respiratory tract (bronchoalveolar lavage fluid, tracheal secretion, sputum, nasopharyngeal secretion, oropharyngeal secretion, etc.) for direct detection of the virus. The test is used for primary laboratory diagnostic examination of patients with suspected SARS-CoV-2 infection. Additionally, there are serological tests for the detection of antibodies against SARS-CoV-2 in the blood. The antibody test is the ideal supplement to direct detection. It supports the diagnosis of SARS-CoV-2 infection and helps to confirm the RT-PCR results. Moreover, the determination of antibodies is relevant for the clarification of suspected cases of SARS-CoV-2 without symptoms or with negative results in direct detection. In addition to their importance for diagnostics, serological tests can also be used to gather epidemiological data and for outbreak control. 

World Health Organisation (WHO) https://www.who.int/

European Centre for Disease Prevention and Control (ECDC) https://www.ecdc.europa.eu/en/home 

EUROIMMUN SARS-CoV-2 tests are CE-marked and can be used for COVID-19 diagnostics. The Anti-SARS-CoV-2 ELISA (IgG) and the EURORealTime SARS-CoV-2 were also approved by the US American Food and Drug Administration (FDA) through an emergency use authorisation (EUA) for use by authorised laboratories (validity of the FDA EUA according to the respective US-specific instructions for use), and by the Brazilian Health Regulatory Agency ANVISA.

Our tests are exclusively performed in the laboratory. The required samples (blood, swabs, etc.) are taken by a physician, e.g. your doctor, and then sent to the respective laboratory in charge.

If you are interested, please get in touch with the Clinical Immunological Laboratory Prof. Dr. med. Winfried Stöcker (Seekamp 31, D-23560 Lübeck).

With the Anti-SARS-CoV-2 ELISAs (IgA and IgG), EUROIMMUN was one of the first companies to offer CE-marked test systems for the detection of antibodies against SARS-CoV-2 already from March 2020. By now, our range of antibody tests for COVID-19 diagnostics has been extended by the CE-marked Anti-SARS-CoV-2 NCP ELISAs (IgG, IgM), the Anti-SARS-CoV-2 QuantiVac ELISA (IgG), the Anti-SARS-CoV-2 Omicron ELISA (IgG), the Anti-SARS-CoV-2 RBD ChLIA (IgG), the EUROLINE Anti-SARS-CoV-2 Profile (IgG) and the surrogate virus neutralisation test SARS-CoV-2 NeutraLISA. For acute diagnostics, in addition to the EURORealTime SARS-CoV-2 and the EURORealTime SARS-CoV-2 Fast for direct detection of the virus via reverse-transcription real-time PCR (RT-PCR), we also offer the combination test EURORealTime SARS-CoV-2/Influenza A/B for differential diagnostics of COVID-19 and influenza as well as the SARS-CoV-2 Antigen ELISA for detection of the viral nucleocapsid protein. The ELISAs, chemiluminescence immunoassay (ChLIA), immunoblot, EURORealTime tests and antigen ELISAs are designed for use in diagnostic laboratories and are exclusively supplied to these.

These tests are not rapid tests for at-home testing.

If you wish to receive a quote for our tests, please contact your local distributor.

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Please note that our self-developed test systems are not rapid tests for at-home testing. They are designed for the investigation of large samples volumes and their processing requires laboratory equipment.

If you suspect to be infected with SARS-CoV-2, please ask your physician for a laboratory diagnostic test.

Suitable methods for the identification of an acute infection are the detection of viral RNA via RT-PCR or that of virus proteins in sample material of the upper respiratory tract. RT-PCR tests enable pathogen detection a few days after virus contact even in subclinical or asymptomatic infections. The virus can be detected up to 14 days after the onset of symptoms. Moreover, by the combination of different parameters, multiparameter RT-PCR tests allow the detection and differentiation of SARS-CoV-2 and other respiratory pathogens, such as influenza viruses, in one reaction. However, as soon as the immune reaction starts and the viral load declines, the sensitivity of this test method decreases as well. The pathogen can then no longer be detected in all patients.

Serology, i.e. the detection of antibodies, expands the diagnostic window over the first one to two weeks. Serology allows identification of persisting, but no longer acute, or past infections with SARS-CoV-2.

Moreover, the S1 domain of the spike protein used in the Anti-SARS-CoV-2 ELISA, Anti-SARS-CoV-2 QuantiVac ELISA, Anti-SARS-CoV-2 RBD ChLIA (IgG), EUROLINE Anti-SARS-CoV-2 Profile and SARS-CoV-2 NeutraLISA contains the receptor binding site (RBD) of SARS-CoV-2, by which the virus binds to human cells. Especially IgG antibodies against the S1 domain/RBD might therefore have a virus-neutralising and thus protective function. These assays can therefore also be applied to support the assessment of the immune reaction after infection or vaccination with one of the COVID-19 vaccines currently approved in the EU, which are all based on the wild type version of the spike protein (Wuhan-Hu-1) (further information here).

Using antibody tests based on the nucleocapsid protein (NCP), antibody reactions after vaccination can be distinguished from those formed after a contact with SARS-CoV-2 (does not apply for the use of vaccines which also contain a N component). A positive result in the Anti-SARS-CoV-2 NCP ELISA (IgG) indicates a past contact with the pathogen. However, even with a negative result a past SARS-CoV-2 infection cannot be excluded.

The assay is suitable for the detection of a past contact with the pathogen. It enables identification of an immune reaction, especially with an unclear antibody status or a missing antibody reaction. Currently, it is assumed that in 10% of symptomatic and 40% of asymptomatic COVID-19 patients, specific IgG antibodies disappear over the course of time and a past infection can only be detected based on the T-cell activity.

According to current knowledge, a T-cell response is also linked to a high immunoprotection even if specific antibodies are absent.

Moreover, the Quan-T-Cell can be used to evaluate the immune reaction following a COVID-19 vaccination. This may be relevant especially for those groups at risk in whom a reduced antibody reaction is to be expected, e.g. immunosuppressed patients.

The WHO estimates an incubation period of 1 to 14 days for SARS-CoV-2. In most cases, it is approximately 5 days. This is the time that passes between the contact with the virus and the onset of first symptoms.

It is assumed that the virus can be detected directly after symptom onset in swabs of the upper respiratory tract by means of RT-PCR or antigen tests.

In viral infections, antibodies are generally only produced at the earliest one week, frequently two weeks after the onset of symptoms and are detectable only then. Specific IgG antibodies can be detected with high sensitivity using for instance the Anti-SARS-CoV-2 ELISA (IgG) or Anti-SARS-CoV-2 QuantiVac, respectively, or the Anti-SARS-CoV-2 NCP ELISA (IgG) from approximately day 10 after symptom onset. IgG antibodies against the nucleocapsid protein (NCP) often occur some days prior to the anti-S1 IgG antibodies. A positive test result confirms virus contact. The Anti-SARS-CoV-2 ELISA (IgA) and the Anti-SARS-CoV-2 NCP ELISA (IgM) are both suitable for early monitoring of an immune response after positive direct detection when specific IgG antibodies have not yet been produced. Neither IgA nor IgM determination are suitable for identification of an acute SARS-CoV-2 infection. The surrogate virus neutralisation test SARS-CoV-2 NeutraLISA is intended for the detection of neutralising antibodies in samples taken at least 15 days after the onset of symptoms.

The T-cell response to SARS-CoV-2 occurs much earlier than the antibody reaction. The activity of specific T-cells can be determined already shortly after onset of symptoms via the release of the signal molecule interferon gamma (IFN-γ).

As long as the virus can be detected in secretions of the respiratory tract by means of reverse-transcription real-time PCR, the patient is assumed to be infectious.

The ELISA technique cannot show whether the detected antibodies have a neutralising effect on the pathogen or not. Whether the IgG results obtained by ELISA provide information on the immunity of a patient is a subject of current scientific research. However, by selection of a suitable antigen that induces the formation of neutralising antibodies, it might be possible to deduce the respective functionality of the antibodies. It can generally be assumed that immunity is mainly associated with class IgG antibodies, which are detected by our Anti-SARS-CoV-2 ELISA (IgG), Anti-SARS-CoV-2 QuantiVac ELISA (IgG), Anti-SARS-CoV-2 RBD ChLIA (IgG) and EUROLINE Anti-SARS-CoV-2 Profile (IgG). These tests are based on the S1 domain of the spike protein, including the immunologically relevant receptor binding domain (RBD) as the antigen. This is also indicated by many published results as, for instance, from the so-called Heinsberg Study by the University of Bonn. The study underlined the high quality of the Anti-SARS-CoV-2 ELISA (IgG) and showed that the ELISA correlated very well with neutralisation assays.

Moreover, EUROIMMUN has developed a surrogate virus neutralisation test which provides semiquantitative determination of neutralising antibodies that inhibit the binding of SARS-CoV-2 S1/RBD to the ACE2 receptors of the human host cells. The results obtained with the NeutraLISA show a very high agreement in the comparison with a plaque reduction neutralisation test (PRNT50).

It is also becoming increasingly clear that not only do antibodies against SARS-CoV-2 play a role in the development of immunity to coronavirus, but that T cells, as part of the cellular immune response, also have an important function in the formation of an immunological memory for SARS-CoV-2. EUROIMMUN’s interferon-gamma Release Assay (IGRA) Quan-T-Cell supports the detection of a T cell immune response following SARS-CoV-2 infection (or vaccination against the virus) by allowing the activity of SARS-CoV-2-specific T cells to be determined from the release of IFN-y.

EURORealTime SARS-CoV-2, EURORealTime SARS-CoV-2 Fast and EURORealTime SARS-CoV-2 Influenza A/B: Yes. Due to the detection of two specific gene sequences of the virus, SARS-CoV-2 infections can be reliably identified and distinguished from other coronavirus or respiratory infections.

Antigen ELISA: During validation of the SARS-CoV-2 Antigen ELISA no cross reactions with antigens of other worldwide occurring coronaviruses were observed. However, due to the close relationship between SARS-CoV(-1) and SARS-CoV-2, cross reactions with the antibodies against SARS-CoV(-1) cannot be excluded.

Anti-SARS-CoV-2 ELISA/Anti-SARS-CoV-2 QuantiVac ELISA/Anti-SARS-CoV-2 NCP ELISA: During validation of the ELISAs, no cross reactions with antibodies against other worldwide occurring coronaviruses were observed. However, due to the close relationship between SARS-CoV(-1) and SARS-CoV-2, cross reactions with the antibodies against SARS-CoV(-1) cannot be excluded.

EUROLINE Anti-SARS-CoV-2 Profile (IgG): During validation of the immunoblot, no cross reactions with antibodies against other worldwide occurring coronaviruses were observed. However, due to the close relationship between SARS-CoV(-1) and SARS-CoV-2, cross reactions with the antibodies against SARS-CoV(-1) cannot be excluded.

Anti-SARS-CoV-2 Omicron ELISA (IgG)*: In the validation of the Anti-SARS-CoV-2 Omicron ELISA (IgG), cross reactions were observed in only 1.2% of the analyses and are thus unlikely. However, cross reactions with antibodies against SARS-CoV(-1) are likely due to the close relationship between the two viruses.

Anti-SARS-CoV-2 RBD ChLIA (IgG): During validation of the ChLIA, no cross reactions with antibodies against other worldwide occurring coronaviruses were observed. However, due to the close relationship between SARS-CoV(-1) and SARS-CoV-2, cross reactions with the antibodies against SARS-CoV(-1) cannot be excluded.

SARS-CoV-2 NeutraLISA: During validation of the assay, no cross reactions with antibodies against other worldwide occurring coronaviruses were observed. However, due to the close relationship between SARS-CoV(-1) and SARS-CoV-2, cross reactions with the antibodies against SARS-CoV(-1) cannot be excluded.

Interferon-gamma release Assay Quan-T-Cell: During validation of the Quan-T-Cell-ELISA in combination with the Quan-T-Cell SARS-CoV-2, no cross reactions with antibodies against other worldwide occurring coronaviruses were observed. However, due to the close relationship between SARS-CoV(-1) and SARS-CoV-2, cross reactions with the antibodies against SARS-CoV(-1) cannot be excluded.

^{*The test also detects antibodies against other SARS-CoV-2 variants.}

EURORealTime SARS-CoV-2, EURORealTime SARS-CoV-2 Fast and EURORealTime SARS-CoV-2 Influenza A/B: The tests are compatible with the standard equipment (real-time PCR thermocycler) present in most of the molecular-diagnostic laboratories. The EURORealTime SARS-CoV-2 was validated on the following real-time-PCR cyclers: 7500 Fast Real-Time PCR Instrument (Applied Biosystems), LightCycler® 480 II (Roche), CFX 96 Touch (Bio-Rad), RotorGene Q (Qiagen) and qTower3 (Analytik Jena). If other cyclers are used, they must be validated by the laboratories themselves. The EURORealTime SARS-CoV-2 Influenza A/B was validated on the following real-time-PCR cyclers: 7500 Fast Real-Time PCR Instrument (Applied Biosystems), CFX 96 Touch (Bio-Rad) and qTower3 (Analytik Jena). If other cyclers are used, they must be validated by the laboratories themselves.

Antigen ELISA: Photometer, incubator or water bath for 37°C. Each test kit contains a 96-well ELISA microplate (individual break-off wells, enabling adjustment to the number of samples to be analysed) and all reagents and control materials required for the test performance. One packaging unit allows the analysis of 93 patient samples. Like other ELISAs, the tests can be performed manually. Automated processing on our EUROIMMUN Analyzer I and I-2P is of course also possible.

Anti-SARS-CoV-2 ELISA/Anti-SARS-CoV-2 QuantiVac ELISA / Anti-SARS-CoV-2 Omikron ELISA / Anti-SARS-CoV-2 NCP ELISA / SARS-CoV-2 NeutraLISA: Photometer, incubator for 37°C. Each test kit contains a 96-well ELISA microplate (individual break-off wells, enabling adjustment to the number of samples to be analysed) and all reagents and control materials required for the test performance. Depending on the number of calibrators, up to 93 patient samples can be analysed with one packaging unit. Like other ELISAs, the tests can be performed manually. Automated processing on our EUROIMMUN Analyzer I and I-2P is of course also possible (Analyzer I: up to 7 microplates, up to 180 samples per test run) and the EUROLabWorkstation ELISA (up to 15 microplates, up to 748 samples per test run).

Anti-SARS-CoV-2 RBD ChLIA (IgG): Can only be processed automatically using the random access instruments IDS-i10 and IDS-iSYS Multi-Discipline Automated System (with software from 15.06a). The possibility of loading the samples continuously allows each patient sample to be processed as a single determination with minimal effort and short reaction times. Each test kit contains one cartridge and ready-for-use reagents for 100 analyses. In addition, the control set Anti-SARS-CoV-2 RBD ChLIA (IgG) as well as further reagents for the random access instruments and a RFID reader are required.

EUROLINE Anti-SARS-CoV-2 Profile (IgG): Rocking shaker (for manual processing); the test kit contains all required reagents and either 16 or 64 membrane strips or pre-equipped incubation trays with 50 strips (depending on the format). The immunoblots can either be conveniently processed using the compact fully automated device EUROBlotOne or alternatively incubated using the EUROBlotMaster and then scanned with a flatbed scanner. In both cases, the immunoblots are evaluated automatically using the EUROLineScan software.

Quan-T-Cell ELISA in combination with Quan-T-Cell SARS-CoV-2: The determination of the SARS-CoV-2-specific T-cell activity requires the stimulation tube set Quan-T-Cell SARS-CoV-2, consisting of 30 x 3 tubes (CoV-2 IGRA STIM, CoV-2 IGRA BLANK and CoV-2 IGRA TUBE), and the Quan-T-Cell ELISA (96-well ELISA microplate) for the quantification of the released IFN-γ. Since three preparations are required for each patient sample and the quantitative ELISA includes 6 calibrators and 2 controls, 29 determinations can be performed per packing unit. Of course the ELISA can be processed completely automatically on our EUROIMMUN Analyzer I and I-2P in combination with the evaluation software EUROLab Quan-T-Cell and the EUROLabWorkstation ELISA.

The validation of the SARS-CoV-2 interferon-gamma release assay Quan-T-Cell was performed using heparinised whole-blood samples. The samples must be stimulated within 16 hours following sample collection. After stimulating the samples and obtaining the plasma, this can be used immediately for analysis or kept in cool storage (long term storage at -20 ° C or short-term storage at 2 – 8 °C).

RNA extraction can be performed by any method (automated or manual) which is suited and validated for the sample materials.
The EURORealTime SARS-CoV-2 test was validated with the QIAamp Viral RNA Mini Kit (Qiagen), the NucleoMag® VET Kit (Macherey-Nagel),  the CMG-2015 Prepito Viral DNA/RNA200 Kit (Chemagen) and the CMG-1033 chemagic Viral DNA/RNA 300 Kit H96 (Chemagen).
The EURORealTime SARS-CoV-2 Fast was validated with the Pre-NAT II NA EU Kit and the CMG-1033 chemagic Viral DNA/RNA 300 Kit H96 (Chemagen).
The EURORealTime SARS-CoV-2 Influenza A/B was validated with the QIAamp Viral RNA Mini Kit (Qiagen), the CMG-1033 chemagic Viral DNA/RNA 300 Kit H96 (Chemagen) and the CMG-2017 Prepito Viral DNA/RNA300 Kit (Chemagen).

Validation of the EURORealTime SARS-CoV-2, EURORealTime SARS-CoV-2 Fast and EURORealTime SARS-CoV-2 Influenza A/B was based on RNA preparations from throat swabs. The EURORealTime SARS-CoV-2 is also validated for the use of saliva as sample material. Other sample materials as RNA sources can be used as well, but have to validated by the customer.

Here, the shipping time of the sample to the laboratory and the laboratory capacities must be taken into account. The sole test performance is around 2 hours with the antibody ELISA, the NeutraLISA or the immunoblot, around 45 minutes with the ChLIA, around 4 hours with the antigen ELISA and around 1.5 hours or 45 minutes (EURORealTime SARS-CoV-2 Fast) with the EURORealTime tests.

Stimulation of the T-cells during the performance of the SARS-CoV-2 interferon-gamma release assay Quan-T-Cell takes up 20 to 24 hours; the subsequent quantification of released IFN-γ by ELISA approximately 4 hours.

The stimulation is based on the SARS-CoV-2 spike protein (see also antigen description of the antibody ELISAs) Both CD8+- and CD4+-T-cells are activated by this.

The Anti-SARS-CoV-2 ELISAs (IgA and IgG) and the Anti-SARS-CoV-2 QuantiVac ELISA (IgG) are based on the S1 domain of the spike protein (S), which is produced in a sophisticated procedure with a human cell line. This procedure allows the presentation of complex three-dimensional structures and post-translational glycosylations. Therefore, antibodies that exclusively react with authentic epitopes of SARS-CoV-2 can also be detected. Due to the use of this antigen, the Anti-SARS-CoV-2 ELISAs (IgA and IgG) and the Anti-SARS-CoV-2 QuantiVac ELISA (IgG) can detect antibodies with high specificity and sensitivity. Moreover, the S1 domain contains the receptor binding site (RBD) of SARS-CoV-2 via which the virus binds to human cells. Especially IgG antibodies against the RBD/S1 domain might have a virus-neutralising and thus protective function:It could be shown that antibodies against S1/RBD effectively prevent attachment of the virus to human cells. The question of immunity is still subject of intensive research.

Accordingly, the currently approved vaccines are based on the use of different presentations of the SARS-CoV-2-specific spike protein. Thus, the S1 antigen plays a crucial role in the detection of antibodies produced following vaccination (more information can be found here).

The Anti-SARS-CoV-2 NCP ELISAs (IgG and IgM) are based on a modified variant of the particularly immunogenic viral nucleocapsid protein (NCP). Antibodies against the nucleocapsid protein are typical markers of infections with SARS-CoV-2. The nucleocapsid protein in its full length, however, presents many homologies within the coronavirus family, which is why the occurrence of unspecific (false positive) reactions with antibodies against other worldwide circulating human pathogenic coronaviruses cannot be excluded. Therefore, instead of the complete protein, the EUROIMMUN Anti-SARS-CoV-2 NCP ELISAs (IgG and IgM) are based on a designer antigen in which unspecific conserved regions were eliminated.

The Anti-SARS-CoV-2 Omicron ELISA (IgG) is based on the recombinant S1 domain of the spike protein of the SARS-CoV-2 Omicron variant.*

For the EUROLINE Anti-SARS-CoV-2 Profile (IgG) the recombinant S1 and S2 domain of the spike protein as well as the recombinant nucleocapsid protein of SARS-CoV-2 were applied to the immunoblot strips as separate antigen bands. The antigen composition enables a differentiated determination of anti-SARS-CoV-2 IgG. The recombinant nucleocapsid proteins of four other human pathogenic coronaviruses (HCoV-229E, HCoV-NL63, HCoV-OC43 and HCoV-HKU1) are also included on the test strip as individual bands and only serve for information purposes.

^{*The test also detects antibodies against other SARS-CoV-2 variants.}

Antibodies develop approximately one to two weeks after the onset of symptoms. The course of the immune response may vary significantly between patients. Usually, antibodies against the viral nucleocapsid protein (NCP) are produced earlier (approximately seven days after symptom onset) than antibodies against the spike protein (S/S1 domain) (approximately 10 days after symptom onset). Antibodies of immunoglobulin classes A (IgA) and M (IgM) often occur earlier than those of class G (IgG). IgA and IgM indicate the starting immune response, while IgG antibodies most probably play a role in the development of immunity. Especially IgG antibodies against the S1 domain of the spike protein may have a virus-neutralising and thus protective function. The question of immunity is still subject of intensive research.

Moreover, in the literature, examples of individual patients have been repeatedly described in whom the secretion of antibodies only started with a time delay of several weeks or failed to occur entirely. These patients are negative for anti-SARS-CoV-2 IgG and reduce the clinical sensitivity of serological tests.

The Anti-SARS-CoV-2 QuantiVac ELISA (IgG) allows quantification of the anti-S1 IgG antibody concentration. The test system has been already calibrated using the NIBSC Working Standard (NIBSC code 20/162), which is the only currently available reference material for anti-SARS-CoV-2 antibodies. As soon as an international standard serum is officially available, it will be possible to issue the results of the Anti-SARS-CoV-2 QuantiVac ELISA (IgG) in IU/ml. The quantification of the anti-S1 antibody concentration plays an important role in the development of S1-based vaccines. Please find further information here.

The SARS-CoV-2 NeutraLISA is modelled upon nature and imitates the natural process of virus neutralisation. The test system exclusively detects antibodies against SARS-CoV-2 that actually inhibit the binding of the viral S1 domain/RBD to the human ACE2 and thus contribute to the neutralisation of the virus. The test results show a very high agreement with those from a plaque reduction neutralisation test (PRNT50). Unlike the classic virus neutralisation assay, however, this ELISA-based test does not require a BSL-3 laboratory, can be easily integrated in the laboratory routine and is automatable even for high throughputs. In this way, the SARS-CoV-2 NeutraLISA makes an important contribution to the evaluation of the immune reaction after infection or vaccination with an S1-based vaccine.

By using the S1 subunit of the spike protein in the Anti-SARS-CoV-2 QuantiVac ELISA (IgG) and Anti-SARS-CoV-2 RBD ChLIA (IgG), antibodies induced by the spike-protein-based vaccines in use (applicable to all COVID-19 vaccines currently approved in the EU) can be detected and quantified to support the assessment of the immune protection achieved through vaccination.

For neutralisation of the virus, a sufficient amount of neutralising antibodies against S1/RBD must have been produced. In a comparison study, an excellent correlation of the Anti-SARS-CoV-2 QuantiVac ELISA with the Neutralization Antibody Test (GenScript cPass) was achieved. It can therefore be assumed that the EUROIMMUN ELISA also detects neutralising antibodies. Moreover, the SARS-CoV-2 NeutraLISA is a system which exclusively detects antibodies against neutralising antibodies. The results from the NeutraLISA also show a very high agreement with the results of plaque reduction neutralisation test (PRNT50).Well-characterised immunisations against other pathogens have shown that protection is only achieved at a certain threshold. This threshold must be able to be reliably and comparably used, independently of the vaccine (provided that a similar antigen is used) and test system. The definition of such a value is based on the results obtained from large-scale studies or long-time observations and is generally established based on evidence. The values must therefore be defined by independent institutions or the manufacturers of vaccines with reference to study results.

Since the clinical studies for the assessment of the vaccination success are still being evaluated, thresholds for the assessment of the antibody activity after vaccination have not yet been determined. However, it can be assumed that the determination of the antibody level will be one of the most important instruments in the assessment of the vaccination success. It is therefore very likely that one or several thresholds will be established. Since all of the approved vaccines are based on the spike protein, these values will probably refer to the IgG response to the spike protein. Due to the very exact quantification of the antibody concentrations with the Anti-SARS-CoV-2 QuantiVac ELISA and Anti-SARS-CoV-2 RBD ChLIA (IgG) the assays can be applied with regard to every prospective threshold that might possibly be used for assessing the vaccination success.

The international standard serum, the “First WHO International Standard Anti-SARS-CoV-2 Immunoglobulin”, was officially released at the end of December 2020. An excellent correlation of the Anti-SARS-CoV-2 QuantiVac ELISA (IgG) with this reference material has already been shown, so that the results of the Anti-SARS-CoV-2 QuantiVac ELISA (IgG) can be given in standardised units (BAU/ml, numerically identical to IU/ml). The antibody concentrations determined using the Anti-SARS-CoV-2 RBD ChLIA (IgG) can also be converted into BAU/ml.

Since the beginning of the pandemic, different SARS-CoV-2 variants were observed. These include Alpha (B.1.1.7 lineage), Beta (B.1.351 lineage) and Gamma (P.1 lineage), which were initially classified as variants of concern (VOC), as well as the variants  Delta (B.1.617.2 lineage) and Omicron (B.1.1.529 lineage), including subtypes, that are currently classified as VOC.

EURORealTime test systems:

Since the EURORealTime SARS-CoV-2, EURORealTime SARS-CoV-2 Fast and EURORealTime SARS-CoV-2/Influenza A/B test systems are based on the ORF1ab and N genes of SARS-CoV-2, they are not affected by the mutations in the S gene. For the above mentioned variants, however, further mutations in other genes of the SARS-CoV-2 genome have been described, which may affect primer and probe binding.  Based on comparisons of the primer and probe sequences with published sequences of  different SARS-CoV-2 variants (e.g. Alpha, Beta, Gamma, Delta and Omicron) all of these variants are expected to be reliably detected with the EURORealTime SARS-CoV-2, EURORealTime SARS-CoV-2 Fast and EURORealTime SARS-CoV-2/Influenza A/B tests.

ELISAs, NeutraLISA, ChLIA, Immunoblot and Quan-T-Cell (IGRA):

Although the variants Alpha (lineage B.1.1.7), Beta (lineage B.1.351), Gamma (lineage P.1) and Delta (lineage B1.617.2) present mutations in the receptor-binding domain (RBD), which is an important component of the S1 domain of the spike protein used as an antigen in the Anti-SARS-CoV-2 ELISA (IgA/IgG), Anti-SARS-CoV-2 QuantiVac ELISA (IgG), SARS-CoV-2 NeutraLISA, Anti-SARS-CoV-2 RBD ChLIA (IgG), EUROLINE Anti-SARS-CoV-2 Profile and Quan-T-Cell SARS-CoV-2, these mutations are not expected to affect the performance of these test systems. First internal studies using samples from patients who have been infected with the Alpha, Beta or Gamma variant, confirm this assumption concerning the antibody test systems. We do not yet have the experimental data to make statements on the possible effects of the Omicron variant on our serological test systems for the detection of antibodies against the S1 domain. The acquisition of the serum samples required from patients positive for the Omicron variant but immunologically naive to other SARS-CoV-2 variants (i.e. who have never been in contact with the wild type form or other variants and were not vaccinated against SARS-CoV-2) proves very challenging. Although it cannot generally be excluded that an infection with the SARS-CoV-2 variant Omicron leads to formation of antibodies which are not or only poorly detectable by the S1 wild type isolate (Wuhan-Hu-1) used by us, our in silico analyses do show that the amino acid sequence of the S1 domain and the RBD within the S1 domain of the Omicron variant and the wild type are highly homologous.

Therefore, we do not expect a negative effect of the Omicron mutations on the performance of our test systems for antibodies against the S1 domain at this stage.

Concerning our Quan-T-Cell, we also do not yet have the experimental data to make statements on the possible effects of the Omicron variant on the detection of SARS-CoV-2-specific T-cell activity. However, a variety of publications already confirm that T-cell reactivity against virus variants including Omicron is very stable and that it offers a certain level of variant-independent immune protection.

Generally, individual amino acid substitutions in an antigen do not significantly alter the properties of the antibody or T-cell binding to the whole protein since only a single epitope is affected. Due to the fact that proteins have a large number of epitopes, the loss of a single binding site can be easily compensated. Furthermore, an individual amino acid substitution does not necessarily mean that antibodies or T cells against a variant can no longer bind to the respective epitope of the original variant. In many cases, no deterioration in the binding behavior is to be expected because, for instance, not every site in the epitope is targeted by the antibodies or T cells.

With respect to the SARS-CoV-2 Antigen ELISA and Anti-SARS-CoV-2 NCP ELISA (IgG, IgM) it is not expected that the new variants affect the performance of the test systems. These tests are based on the detection of the viral nucleocapsid protein and human antibodies (IgG, IgM) against this protein, respectively, and are thus not affected by the mutations in the S gene.

EUROIMMUN carefully follows all reports on new SARS-CoV-2 variants from sources such as ECDC, WHO or FDA. Possible influences of new variants on our tests systems are investigated immediately. Based on these and other analyses of, for instance, current literature, reports from health authorities and feedback from laboratories, a reassessment of the situation is carried out regularly as part of risk management in order to be able to react with short notice if necessary and take preventative or corrective measures where required.