Show me your DNA

Galina Kostina, "Expert", 06/16/2008

In the 1997 film "Gattaca", the events of which take place in the world of advanced DNA technologies, a nurse shocks a happy mother with a newly born baby by reading the diagnosis: the risk of nervous diseases is 60%, the tendency to manic depression is 42%, loss of clarity of perception is 89%, complications of the cardiovascular system - 99%, estimated life span of 30.2 years. The child was illegitimate: his genome was not subjected to total reprogramming at the time of conception.

Now, ten years later, the film no longer seems so fantastic. In any case, DNA diagnostics has entered everyday life. And it all started on one of the April nights of 1983, when American scientist Carrie B. Mullis came up with a polymerase chain reaction (PCR) on the way to his dacha in Northern California, for which he received the Nobel Prize ten years later. The method, which made it possible to obtain billions and trillions of copies of given pieces of DNA or RNA, opened up the widest prospects for research purposes, as well as for medical diagnostics; in particular, it made it possible to quickly and accurately determine the presence of many infectious agents in a blood sample. Shortly after the discovery of PCR, devices for this reaction began to be made in the world. Oddly enough, they immediately began to make them in Russia. Moreover, we have achieved serious success – the DNA Technology company, established in 1993, has been involved in the operation of a landmark world discovery and today sells PCR equipment for $ 10 million a year.

Without pipetteYuri Trofimov worked at the Institute of High Energy Physics, and his son Dmitry Trofimov worked at the SSC "Institute of Immunology".

He successfully got into the laboratory of human immunogenetics, which, in particular, dealt with the genes responsible for tissue compatibility, which is important for transplantation. A wise leader, excellent equipment – the last gift of socialism, the opportunity to do research and travel around the world. The idea to deal with devices and reagents for PCR came from Dmitry. Not only in the West, but also in Russia, they began to use the PCR method without special devices, because it turned out to be so simple that at first its creator himself was surprised, and then other scientists who asked a painful question for every scientist: why didn't I think of it before? It was simply necessary to heat and cool a test tube containing a DNA or RNA sample along with certain reagents according to a certain program. The method attracted by its effectiveness. For example, you need to determine if there is a herpesvirus in the sample. There are several types of analyses with which you can do this. But if there are not so many viral particles, then they may not appear. PCR allows us to stamp such a huge number of specific pieces of virus DNA (characteristic only for it) that it will be impossible not to notice it. In addition, it is done in a couple of hours.

Soon the first devices began to appear in the West – the so–called amplifiers (amplification - an increase in the number of copies of DNA), or, in another way, thermocyclers. They only carried out the reaction itself, in which the desired specific fragment of DNA or RNA was built up. It was not difficult for the team of Trofimov Sr. to come up with such a simple device without even looking at the samples that began to be made abroad. In addition, copying for specialists of such a high class seemed simply unacceptable. The device turned out not to be painfully beautiful, but simple, reliable and cheap. The production was organized in the premises of one of the enterprises in Protvino.

Creating the first device was the first step. The test tubes with samples had to be removed from it and then by the traditional method of electrophoresis to determine whether the reaction had passed and whether the desired virus was in the sample.

"The main advantage of the PCR method – its high sensitivity – at that moment became its Achilles heel," says Dmitry Trofimov. – For the trout, you need to remove a drop of material from the test tube with a pipette. In an open space, the smallest fragments could fall into other samples. False diagnoses have been made. It was a real threat to the DNA diagnostic method." The training of laboratory personnel began, but, fortunately for the staff, a new method based on fluorescent labels appeared in the world. These tags were called molecular beacons: they made it possible to immediately see if the reaction had passed, if it had passed, the beacon would light up.

By the end of the 90s, devices that performed PCR and detection using fluorescent tags in real time began to appear in the world. The first such devices, according to Dmitry Trofimov, cost about 100 thousand dollars. Russian diagnostic laboratories, which used an economical amplifier and electrophoresis, which was not the most suitable for PCR, had a problem where to get so much money for a foreign device.

"We solved this problem quite quickly and successfully,– says Dmitry Trofimov. "Our team of engineers has created another unique device that carried out detection with fluorescent tags." True, in the Russian version, the test tube had to be removed from the amplifier, but there was no need to open it: it was inserted into a fluorescent detector, and the beacon signaled the answer. Two devices – a thermal cycler and a fluorescent detector – cost three to four thousand dollars instead of one for 100 thousand, and then the choice was obvious. "Gin" was so successful that a few years later its analogues began to appear on the market.

But foreign devices not only gave an answer whether there was a virus or not, but also determined the number of viral particles. They were called real-time amplifiers. "The mechanism is based on the knowledge of the formula by which the genetic material of the infectious agent increases during the reaction," says Denis Rebrikov, Deputy General Director for Science at DNA Technology. – At each temperature cycle, the number doubles, and so on exponentially. Based on the accumulated amount, according to this formula, it is possible to determine how many particles were in the test tube before the reaction began. "

When the company took up the creation of the real-time device after "Gin", it was decided not to save money, as it was at the very beginning of the journey. "We made the first devices at a time when the country was tight with finances, our customers, mostly newly appeared private medical centers, clearly monitored the principle of price-quality in order to equip themselves as much as possible, but for moderate money," says Dmitry Trofimov. – The new device could no longer be cheap, because it was more high-tech, it required a completely different filling, part of which had to be bought abroad. In addition, I wanted to make not just a functional piece of iron, but a device that is not inferior to Western analogues either in filling or in appearance. And by some indicators even surpassing them." In particular, the new device, unlike the Western ones, had a program that allows you to identify genetically modified products and count the number of GM ingredients.

Having installed new devices in their laboratory next to the brand-new American ones, the heads of DNA Technology noted with satisfaction how quickly the staff switched to Russian models. "We are both producers and consumers, because we have our own laboratory. It is very comfortable. We immediately get an assessment of our products, we change something in the programs for the convenience of medical staff," Dmitry Trofimov continues. – It is important that all programs are in Russian. It is important that we conduct staff training, specialists from many laboratories come to us for courses. And it is important that we have a staff of engineers who carry out maintenance. No foreign company can boast of this."

The company makes not only devices, but also various test systems for PCR. At the same time, initially the devices, unlike Western ones, were produced as open systems – they could use not only reagents made by the company itself, but also by other firms. DNA Technology produces additional equipment with which it is possible to fully equip the laboratory for PCR. Another unique product is a mobile PCR laboratory for field work, which one person can prepare for work in ten minutes. It was composed for the love of compactness. Offered to the military and the Ministry of Emergency Situations. Individual divisions are buying, but there are no large orders yet.

Difficulties of innovative businessIn the early 90s, the creators of the company did not build business plans.

It was necessary to find a source of income that would support not only pants, but also scientific ambitions. "In the USSR, scientists satisfied their curiosity at the expense of the state, in the modern economy they often have to pay for it themselves," says Denis Rebrikov. – The company spends a significant part of its budget on scientific (including fundamental) research. The employees of our division, about fifty people, have much less problems in terms of supply than most scientists at the Research Institute. We have good equipment and reagents. The researcher can always order and buy the missing materials. But the quality and efficiency of development largely depends on the infrastructure and the speed of obtaining the necessary reagents." Employees of the company are actively engaged in research at the cutting edge of science, are published in well-known scientific publications, defend dissertations.

The developing and growing market was so good that with its high-quality, but much cheaper than foreign devices, the company did not need to make much effort to sell them. The share of DNA Technology in the Russian market of the first simple thermal cyclers was up to 80%. The share of unique fluorescent detectors "Jin" is 70%. New detecting thermal cyclers occupy about 15% of the market so far. "Here we entered the market late,– admits Dmitry Trofimov. – Now we will catch up with the help of a very competitive version of this device."

In recent years, the company's executives have had to pay much more attention to business management and the market. "Money has appeared in the country, and the consumer no longer counts pennies," says Dmitry Trofimov. – The structure of the market is also changing. If ten years ago the most active consumers were private medical centers, now budget-funded medical institutions have begun to be decently funded. It's a shame, but we are faced with officials' distrust of an innovative Russian product. They say that foreign a priori is better, especially since there is no shortage of funds now. Domestic, according to officials, should be at least ten times cheaper to attract attention. But high-tech equipment can't be much cheaper. There is practically nothing to save on – neither on renting premises, nor on wages, besides, some parts have to be bought abroad."

Now a lot of time is spent on establishing connections, persuasion, participation in tenders. Undeveloped innovation infrastructure also hinders development. In Russia, the company has to create a subsistence economy and do everything itself – from the developer's idea to soldering chips and painting cases. In the West, there are companies that specialize in small-scale production for innovative companies. In Russia, such outsourcing practically does not work. "We are getting into a vicious circle," says Trofimov. – If there are not so many orders from the developers, then such an infrastructure is not being created. And if it's not there, then we slow down a lot." Now in the West, genetics and DNA diagnostics are becoming extremely attractive for investors, huge resources are being concentrated in companies associated with these areas. At the same time, a Russian company will not be able to go toe-to-toe with foreign companies alone. It is necessary to cooperate with other innovative companies or the state. Then the company will be able to grow as a manufacturer of competitive PCR devices. If not, it will remain a small innovative company that will look for its niches in the market of original medical equipment. Dmitry has no doubt about the latter.

Tendency to diabetes and sprintingAnother direction is the development of laboratory services, which initially acted in DNA Technology as an auxiliary object for testing equipment.

However, its services have become more and more in demand in recent years and began to occupy about 20% of the revenue structure (in 2007 – about $ 10 million). But this share may grow. Denis Rebrikov shows a cute box in which a brochure and a test tube are packed. Such a box can already be bought in some pharmacies in England. You open the test tube, soldered into the lid with a thin scraper, take a sample behind your cheek, close the test tube, seal it in an envelope and send it to the appropriate laboratory for DNA analysis. "Scientists have discovered genes that are quite clearly associated with certain diseases," says Denis Rebrikov. – The occurrence of oncological, neurodegenerative, many cardiovascular diseases, diabetes and others largely depends on what changes in the genes contain human cells. In principle, you can do hundreds of tests. How to interpret the results is another matter. Now there are not so many parameters on which it is possible to draw unambiguous conclusions." But even a rather vague result like "your risk of cardiovascular diseases is 10% higher than in the population" can correct the behavior of a person who cares about his health.

Now they are trying to identify by genes not only the presence of diseases or a tendency to them, but also, for example, a tendency to certain types of loads, which is important for the preparation of athletes. "If earlier the coach selected the kids by eye, and it's very good if he had one "goldfish" in his team, then with genetic analysis he can select twenty potential "goldfish" at once and work with them," says Denis Rebrikov. A lot of research in world laboratories is devoted to the connection of genetics with intellectual abilities. But so far, according to Rebrikov, no one can predict by genes whether a child will be a famous chess player, pianist or Nobel laureate in physics.

The field of genetic research is the mainstream of world biology. With new discoveries, the knowledge base will be constantly updated, analyzed, algorithms will appear that can reduce thousands and hundreds of thousands of parameters to specific diagnoses. The list of areas of research using PCR will grow in the same geometric progression as the DNA fragment in the reaction itself. "This market is growing even now, but after a while there may be explosive growth," Mr. Rebrikov believes, "What now seems fantastic will become routine analyses."

Portal "Eternal youth" www.vechnayamolodost.ru16.06.2008