MICRO TECHNOLOGIES IN EXPRESS DIAGNOSTICS OF TOXIGENIC CORYNEBACTERIUM DIPHTHERIAE STRAINS


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Abstract

Aim. Development of micro technologies based approach for express diagnostics of toxigenic C.diphtheriae strains. Materials and methods. Corynebacterium diphtheriae 10648 (tox+) and C.diphtheriae NCTC 10356 (tox-) from Central Health Laboratory (London) reference strains were used as positive and negative controls respectively. Diagnostic kit was created by using fractions of antibodies with high avidity that were obtained by consecutive fractioning of positive antitoxic blood sera and then loaded onto polyacrylamide latex particles with the diameter of 0.81 μm. 20 Elek test positive C.diphtheriae strains and 20 tox gene PCR negative C. diphtheriae strains (i.e. non toxigenic) (Pasteur Research Institute of Epidemiology and Microbiology) were used as control. Indirect hemagglutination with anti-diphtheria antibody diagnostic kit was used as a quantitative control. PCR, Elek test and ICS test were used as quality control. Results. The diagnostic kit obtained had specificity of 97%, sensitivity of 98%. Specimen preparation time is 15-20 minutes, reaction time — 2-3 minutes, and up to 93 specimens can be analyzed on a single microchip. Conclusion. The developed approach has high sensitivity and specificity, is easy to use, and fast in regard to prepara- tion and reaction time. Portability of the apparatus allows the use of reagents in micro volumes.

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МИКРОТЕХНОЛОГИИ В ЭКСПРЕСС-ДИАГНОСТИКЕ ТОКСИГЕННЫХ ШТАММОВ СORYNEBACTERIUM DIPHTHERIAE
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About the authors

L. A Kraeva

Pasteur Research Institute of Epidemiology and Microbiology, St. Petersburg, Russia

T. M Zimina

State Electrotechnical University, St. Petersburg, Russia

G. Ya Tseneva

Pasteur Research Institute of Epidemiology and Microbiology, St. Petersburg, Russia

A. V Solovyev

State Electrotechnical University, St. Petersburg, Russia

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Copyright (c) 2011 Kraeva L.A., Zimina T.M., Tseneva G.Y., Solovyev A.V.

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