Development and testing of a DNA microarray for identification of particularly dangerous infectious pathogens


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Abstract

The continuous necessity for monitoring particularly dangerous infectious diseases determines the urgency of developing new effective methods to identify the pathogens of these diseases. Objective. To design and test a reagent kit involving a DNA microarray to reveal the pathogens of anthrax, plague, tularemia, cholera, legionellosis, and brucellosis. Subjects and methods. The oligonucleotide probes carrying the amino groups at the 5'-end were applied to the surface of aldehyde-coated slides with a noncontact printing plotter. After DNA extraction from inactivated cultures, the investigators used multi-primer PCR, then transcription, by including a RNA-detecting label. Transcription products were hybridized using the DNA microarray, then a laser scanner was applied to analyze fluorescence profiles. Results. The reagent kit involving a DNA microarray has been designed to reveal the pathogens of particularly dangerous infectious diseases, which allows 48 samples to be analyzed. The analysis time is 5.5 hours. Conclusion. The reagent kit has been found to be sensitive and specific, which makes possible using it at the regional laboratories and reference centers.

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About the authors

E. A Pudova

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare; Research Institute of Occupational Medicine, Russian Academy of Sciences

Email: el_pudov@mail.ru

T. A Chekanova

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare; Research Institute of Occupational Medicine, Russian Academy of Sciences

Email: tchekanova74@mail.ru

M. L Markelov

Research Institute of Occupational Medicine, Russian Academy of Sciences

Email: mikhailmarkelov@gmail.com

V. G Dedkov

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare; Research Institute of Occupational Medicine, Russian Academy of Sciences

Email: vgdedkov@yandex.ru

N. P Kirdyashkina

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare; Research Institute of Occupational Medicine, Russian Academy of Sciences

Email: nkirdyashkina@mail.ru

I. P Karaseva

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare

Email: irkesha@list.ru

A. I Sazhin

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare

Email: alexey.sazhin@gmail.com

T. S Zatsepin

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare

Email: tsz@yandex.ru

D. V Utkin

Russian «Microbe» Plague Control Research Institute, Russian Inspectorate for the Protection of Consumer Rights and Human Welfare

Email: rusrapi@mikrobe.ru

N. A Osina

Russian «Microbe» Plague Control Research Institute, Russian Inspectorate for the Protection of Consumer Rights and Human Welfare

Email: rusrapi@mikrobe.ru

S. A Shcherbakova

Russian «Microbe» Plague Control Research Institute, Russian Inspectorate for the Protection of Consumer Rights and Human Welfare

Email: rusrapi@mikrobe.ru

G. A Shipulin

Central Research Institute of Epidemiology, Russian Inspectorate for Protection of Consumer Right and Human Welfare

Email: german@pcr.ru

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