14th ICID - Poster Abstracts - International Society for Infectious ...

When citing these abstracts please use the following reference:
Author(s) of abstract. Title of abstract [abstract]. Int J Infect Dis 2010;14S1: Abstract number.
Please note that the official publication of the International Journal of Infectious Diseases 2010, Volume 14, Supplement 1
is available electronically on http://www.sciencedirect.com
Final Abstract Number: 75.021
Session: Diagnostics
Date: Friday, March 12, 2010
Time: 12:30-13:30
Room: Poster & Exhibition Area/Ground Level
Type: Poster Presentation
Combination of PCR and electrical microarray allows rapid and sensitive multiplex detection of
mosquito-transmitted pathogens
M. Kaiser 1 , M. Ulrich 1 , A. Löwa 2 , G. Piechotta 3 , R. Wörl 4 , H. Ellerbrok 2
1 GenExpress, Berlin, Germany, 2 Robert Koch Institute, Berlin, Germany, 3 Fraunhofer Institut für
Siliziumtechnologie, Itzehoe, Germany, 4 AJ eBiochip GmbH, Itzehoe, Germany
Background: More than 300 million people are infected every year with pathogens transmitted
by mosquitoes, with 3.3 billion at risk. The majority of cases are infections with different
Flaviviridae, Chikungunya Virus or Plasmodia. Standard tests to detect these pathogens are
serological assays like IgM and IgG ELISAs for viral infections and the blood smear test in
malaria diagnosis for the detection of Plasmodia. An early, fast and specific diagnosis often is
vital for efficient treatment of these infections, but serological tests usually remain negative during
the viremic and early symptomatic phase and in addition antibodies show broad cross-reactivity
between Flaviviruses. While smear tests are a cheap and rapid method for diagnosis of malaria
the limited sensitivity of this test does not allow detection of low level Plasmodia infections.
Methods: Real-time RT-PCR is a diagnostic tool with high specificity and sensitivity but has
limitations in the parallel detection of multiple pathogens. Micro arrays tend to be less sensitive
but have the potential to detect and discriminate a brought range of pathogens.
Results: We developed and optimized a highly sensitive multiplex PCR approach designing a
combination of PCR primers for parallel amplification of Dengue viruses, Yellow Fever Virus,
West Nile Virus, Japanese Encephalitis Virus, Chikungunya Virus and Plasmodia. Amplicons are
differentiated on an electrical microarray. Specificity and sensitivity of the multiplex assay was
determined and compared to specific real-time RT-PCR assays.
Conclusion: Our results demonstrate that combination of a broad PCR approach with
differentiation on the electrical microarray is a simple, specific, and sensitive tool for early
differential diagnosis for the majority of mosquito-transmitted tropical fevers.