Estonian researchers creating a new virus analysis

Kadri Tammepuu
, Meie Eesti toimetaja
Mart Ustav.
Mart Ustav. Photo: Sille Annuk

The coronavirus SARS-CoV-2 analyses currently used are too cumbersome and expensive, as well as inaccurate in certain circumstances, says Academician Mart Ustav, Professor of biomedical technology at the University of Tartu institute of technology and the founder and leader of Icosagen Grupp.

Why does the coronavirus analysis take several hours?

The analysis consists of four stages. First they take a sample from the nose with a cotton stick and take it to the lab. Then they remove the genetic material, in this case RNA, from which the DNA copy will be made. The fourth stage will use the polymerase chain reaction to boost the genetic data received from the sample so as to read it as positive or negative from the analysis result.

What are the drawbacks of the present analysis method?

The analysis can prove that the cold, cough and fever were caused by the coronavirus. But the development of the symptoms takes time and therefore the analysis does not provide the answer immediately after infection. If the human immune system discovers a virus in the body, it will activate and an allergic reaction results; the result of the coronavirus is initially fever, rhinitis or cough. People will suffer and die if their immune system reacts too violently and will harm the person’s own organs, e.g. lungs, while combating the virus.

Moreover, the present analyses can provide both false positive and false negative results. In case of people with the initial stage of infection when the viruses have not yet massively multiplied in the body, the analysis need not show the virus. On the other hand, in case of those whose immune system has killed virus and who no longer have the infecting virus, the result may be false positive, since the analysis can establish pieces of the virus’ genomic RNA and the symptoms may still be present.

They also offer rapid tests, which are also inaccurate. What is their problem?

The rapid analyses used at present measure the IgG and IgM antibodies in the person’s blood, which have developed in response to the virus, but they cannot detect the virus itself. The antibodies suitable for detecting this virus still have to be developed.

And this is what you are doing. What kind of methods would be the most suitable for determining the coronavirus?

There are two opportunities. First, studying the immune response which is developing in an infected person. For example, we want to identify four anti-coronavirus protein molecules IgM, IgA, IgG and IgE and their amount in blood, plasma, serum or tissue fluids, because their amount is related to the stage of infection and the state of the organism. The amount of antibodies gives an idea of the organism’s ability to drive out the virus. Eventually the person’s immune response to the virus is the only way of preventing the corona virus disease, since this virus grows only in our cells and of course in those of the bat. A drop of blood taken from the finger could prove that the person carries the virus if the blood sample displays a definite response to the proteins of the virus.

Another opportunity is to determine the virus directly – to determine the virus proteins or antigens from the same sample – from shavings or wash of the mucous membranes.

We are trying to develop analysis methods of both types, which would be accurate, fast and reasonably priced.

The EU Innovative Medicines Initiative IMI2 opened a round of applications for diagnostics and treatment of SARS-CoV-2. Will you take part in it?

Yes, we shall apply for a grant with our foreign partners for the development of diagnostic and treatment methods. But applying for grants is like fishing; you need not catch every time, we do not know what the competitors are offering and whose proposals will be selected. The deadline for applications is March 31.

The world is waiting for faster and more accurate analysis methods. Is the timetable for creating the diagnostic tests already made up?

The projects have been planned for three years together with intermediate goals. Our experience of development with diagnostic antibodies allowing the detection of e.g. heart attack or sepsis give us ground to believe that by summer we shall have lab-grown purified virus proteins or antigens which can be used in diagnostic tests and further development. We also hope by summer to obtain the blood of a patient recovered from the viral infection, from which we can isolate the mononuclear cells and discover the antibodies produced by the organism, which could be suitable for use in diagnostics and treatment. We shall also develop antiviral immunity molecules using chicken and rabbits. The first antibodies should be produced and characterized by autumn.

What would be the probable cost of your future rapid analysis of the coronavirus?

I estimate that the cost of our analysis would be approximately one euro, while that of the analysis currently used in Estonia is roughly 100 euros – the Roche analysis kit costs 80 – 85 euros plus the cost of protection gear and logistics.

How quickly would your analysis provide the answer?

In the ideal case the result would take 20 minutes, while it presently takes, dependent on logistics, up to 24 hours. We are cooperating with a partner whose device, thanks to modern technology, can identify the indications rapidly and accurately. The key components in this work are the virus proteins and sensitive, quick-response antibodies.

You also participate in the grant application.

We have three directions in the treatment opportunities project. First the antibodies, which have the effect of neutralizing the virus, secondly intracellular drugs and thirdly cell therapy.

We have cooperation partners in all three directions in Estonia, Europe, the USA and Canada. We are testing the developed applications on monkeys, because our French partner has a large colony of macaques and they also have the model of the spreading of the coronavirus. We can place our therapeutic antibodies in the model and make sure that they actually present the virus.

When did you begin to think about the analysis and treatment methods of the new coronavirus?

As soon as the genome sequence of the virus was published, just like the other labs all around the world. Everyone understands that the problems of the coronavirus analysis and treatment need to be solved quickly.

How would the daily work of your enterprise change if the grant applications should be successful?

We usually produce proteins for pharmacy companies for developing drugs against various diseases, but we also make therapeutic and diagnostic antibodies. Thus the current projects will continue. If the grant applications should be successful, we shall be able to reorganize our work and hire more staff. A lot of work with the coronavirus analysis methods has already been done and we shall carry on regardless of receiving the grant or not, since we can see a market for these products.

You are writing the project applications in addition to the daily work?

We have a development department and action plans; of course, the priorities can change dependent on the grant application outcomes.

If the virus should mutate by the next season, you would have to change the created diagnostic and treatment methods.

No one knows what will happen to the SARS-CoV-2 virus. But if the know-how is there, we shall solve the same problems much more efficiently and cheaply in the future. The previous SARS came, killed 800 people and vanished.

Now the new virus came from the bats. Maybe we shall get rid of it just like the Chinese did. It is certain that another virus hazardous to people will come, but we do not know when.

Does the Estonian state support your development activities?

We have previously received support from Enterprise Estonia (EAS). There are no separate measures in Estonia for the coronavirus. If the state should order something we would naturally be prepared to provide it.

How much money does one development project require?

The development of an average diagnostic method into a product suitable for the end user costs approximately one million euros. Support would speed up our work. Already halfway there would be intermediary products – proteins, antibodies, new methods – which could be sold to earn money and thus cover part of the development costs. Our market is the whole world, not just Estonia.