S (2012) Rapid and specific detection of Lassa virus by reverse transcription-PCR coupled with oligonucleotide array hybridization

To facilitate sequence-specific detection of DNA amplified in a diagnostic reverse transcription (RT)-PCR for Lassa virus, we developed an array featuring 47 oligonucleotide probes for post-PCR hybridization of the amplicons. The array procedure may be performed with low-tech equipment and does not take longer than agarose gel detection. The method of choice for rapid detection of Lassa virus inblood is reverse transcription (RT)-PCR (2, 4, 7). However, the high genetic variability of the virus (1) prevents the design of probe-based real-time RT-PCR assays that reliably detect all virus strains. Current PCR assays for Lassa virus are based on agarose gel or real-time Sybr green detection. As both ap-proaches are not sequence specific, amplified DNA is se-quenced to confirm the PCR diagnosis. We have developed an oligonucleotide array for sequence-specific post-PCR hybrid-ization of the amplicons as a rapid alternative to sequencing. The array chip is the size of a photo slide and can be scanned with a slide scanner or evaluated with a magnifying glass (Fig. 1A). It was used in conjunction with an existing RT-PCR assa