Real time RNA transcription monitoring by Thiazole Orange (TO)-conjugated Peptide Nucleic Acid (PNA) probes: norovirus detection

Thiazole Orange (TO)-conjugated Peptide Nucleic Acid (PNA) probes have been reported as a valuable strategy for DNA analysis; however, no investigations targeting RNA molecules and no comparisons between different derivatization approaches have been reported so far. In this work, two TO-conjugated PNAs for genogroup II noroviruses (NoV GII) detection were designed and synthesized. Both the probes target the most conserved stretch of nucleotides identified in the open reading frame 1-2 (ORF1-ORF2) junction region and differ for the dye conjugation strategy: one PNA is end-labelled with the TO molecule tethered by a linker; the other probe bears the TO molecule directly linked to the PNA backbone, replacing a conventional nucleobase. The spectroscopic properties of the two PNA probes were studied and their applicability to NoVs detection, using an isothermal assay, was investigated. Both probes showed good specificity and high fluorescence enhancement upon hybridization, especially targeting RNA molecules. Moreover, the two probes were successfully employed for NoVs detection from stool specimens in an isothermal-based amplification assay targeting RNA 'amplicons'. The probes showed to be specific even in the presence of high concentrations of non-target RNA.     

Ultrastructure and Comparative Morphology of Mouth-part Sensilla in Ground Beetle Larvae (Insecta, Coleoptera, Carabidae)

Labial and maxillary palps in 22 species of ground beetle larvae revealed the presence of four different types of sensilla: sensilla digitiformia, sensilla campaniformia, sensilla basiconica and sensilla chaetica. Using transmission electron microscopy techniques, the role of most of them was defined: sensilla campaniformia and sensilla chaetica are mechanoreceptors; while sensilla basiconica are chemoreceptors. Sensilla digitiformia can be both mechanoreceptors and chemoreceptors, though hygroreception may also be postulated on the basis of their abundance on the mouthparts of some hygrophilous species that depend on damp or wet patchy biotopes/seasons. The number and location of the sensory structures has been found to vary among the investigated taxa. The differences are related to the larval behavioural types, and are an adaptive response to different lifestyles, feeding habits and use of space.