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Nucleic Acids Research, 2003, Vol. 31, No. 22 e146
© 2003 Oxford University Press

Article

Viral and microbial genotyping by a combination of multiplex competitive hybridization and specific extension followed by hybridization to generic tag arrays

Baback Gharizadeh, Max Käller, Pål Nyrén, Anders Andersson, Mathias Uhlén, Joakim Lundeberg and Afshin Ahmadian*

Department of Biotechnology, Stockholm Center for Physics, Astronomy and Biotechnology, Royal Institute of Technology (KTH), Roslagstullbacken 21, SE-106 91 Stockholm, Sweden

*To whom correspondence should be addressed. Tel: +46 8 5537 8333; Fax: +46 8 5537 8481; Email: afshin{at}biotech.kth.se

Detection and identification of microbial pathogens are important for disease diagnosis, treatment and prophylaxis measurements. By introducing an innovative technique, we show a robust, reliable and accurate microarray-based method for identification of microbial pathogens. The technique utilizes a unique combination of multiplex competitive hybridization, which enhances hybridization accuracy of oligonucleotides to the specific target, and apyrase-mediated allele-specific extension, which improves specific extension. As a model system, different clinically relevant human papillomaviruses were selected for this study. The method generated accurate results and proves to be promising for specific and correct microbial and viral typing.


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