Development of fast and sensitive protocols for the detection of viral pathogens using a small portable convection PCR platform.

One of the most crucial steps for preventing viral pandemics is the early detection of the causative virus on site. Various molecular and immunological approaches have been developed for virus detection. In this study, we investigated the utility of the recently introduced convection polymerase chain reaction (cPCR) platform for the rapid and sensitive detection of various animal viruses in the field, including the foot-and-mouth disease virus (FMDV) and avian influenza viruses (AIVs). Primer sets were designed to simultaneously detect two highly conserved regions of the FMDV, including the 5' untranslated region (5'-UTR) and 3D gene, and to specifically amplify the NP and hemagglutinin (HA) genes of H5 and H9 subtypes of AIVs. The portable cPCR system was able to amplify from as low as 1 to 10 copies of viral cDNAs in the singleplex mode and 10 to 100 copies of viral cDNAs in the duplex mode within 21 min. Thus, our data suggest that the cPCR protocols developed in this study are highly sensitive and enable quick detection of animal viruses in biological samples.

Data for the optimization of conditions for meat species identification using ultra-fast multiplex direct-convection PCR.

This article contains data related to the research article entitled "Ultra-fast DNA-based multiplex convection PCR method for meat species identification with possible on-site applications" (Song et al., 2017 [1]). Direct PCR that does not require prior DNA extraction is critical for ultra-fast molecular detection of meat species. We successfully acquired DNA by swab sampling in Taq DNA polymerase buffer. To reduce DNA sample preparation time, proteinase K incubation (0.2 µg/mL) and heat inactivation times were decreased to 10 min and 1 min, respectively. The analysis of swabbed DNA samples from mixed meat could differentiate meat species within the mixed sample. The swabbed DNA samples could be diluted 100 times without losing detection sensitivity.

Ultra-fast DNA-based multiplex convection PCR method for meat species identification with possible on-site applications.

The aim of this study was to develop an ultra-fast molecular detection method for meat identification using convection Palm polymerase chain reaction (PCR). The mitochondrial cytochrome b (Cyt b) gene was used as a target gene. Amplicon size was designed to be different for beef, lamb, and pork. When these primer sets were used, each species-specific set specifically detected the target meat species in singleplex and multiplex modes in a 24min PCR run. The detection limit was 1pg of DNA for each meat species. The convection PCR method could detect as low as 1% of meat adulteration. The stability of the assay was confirmed using thermal processed meats. We also showed that direct PCR can be successfully performed with mixed meats and food samples. These results suggest that the developed assay may be useful in the authentication of meats and meat products in laboratory and rapid on-site applications.