Sensitive Combined Assay Detects Multiple Viruses

A combination assay has been developed that identifies several European Arboviruses that cause aseptic meningoencephalitis or febrile disease.

A combined multiplex ligation-dependent probe amplification and flow-through microarray assay was devised for the detection of European Bunyaviruses.

Scientists at the University Medical Center (Göttingen, Germany) decided to try an amplification and hybridization approach, which amplifies a synthetic signal molecule, which is dependent on sequence-specific hybridization rather than an amplification technique that amplifies the target molecule itself. Their methodology combines multiplex ligation-dependent probe amplification (MLPA) with a flow-through chip hybridization system.

The authors developed the assay based on the combination of real time-MLPA (RT-MPLA) and flow-through three-dimensional (3D) universal microarray for the detection of seven European Bunyaviruses, for an expanded accurate diagnosis of aseptic meningoencephalitis and fevers of unknown origin in Europe. The viruses studied were Toscana virus, (TOSV), sand fly fever Sicilian virus (SFSV), sand fly fever Naples virus (SFNV), mosquito-borne Tahyna virus (TAHV), Inkoo virus (INKV), Batai virus (BATV) and tick-borne Uukuniemi virus (UUKV).

Reagents for MLPA and RT-MLPA were obtained from MRC-Holland (Amsterdam, The Netherlands) and the flow-through chip hybridization system was acquired from PamGene ('s-Hertogenbosch, The Netherlands). The sensitivity of MLPA or RT-MLPA flow-through microarray assay ranged from 10 copies for TOSV, INKV, BATV, and UUKV to 1,000 copies for SFSV, SFNV and TAHV. In most cases, the sensitivity of both methods was comparable.

The authors concluded that combined multiplex MLPA and 3D universal microarray assay is sufficiently sensitive and specific for the detection of European Bunyaviruses and show good correlation to monoplex real-time polymerase chain (RT-PCR) assays. The authors expect that this method can be developed into a diagnostic tool for the rapid analysis of infections due to European Arboviruses. The study was published in ther January 2012 issue of Molecular Biotechnology.

Related Links:
University Medical Center
MRC-Holland
PamGene