The following QIAcuity Software Suite Volume Precision Factor (VPF) patches have been released to enable compatibility with VPF file version 6.0 or higher for QIAcuity Software Suite versions 2.0.20. The outstanding stability, even after extended storage at room temperature without the use of any cooling agent, makes the QIAcuity Probe PCR Kit ideal for high-throughput reaction setup and plate-stack handling. The QIAcutiy PCR mixes can be stored at 30☌ for up to 100 hours without impairing the performance of subsequent reactions. Moreover, the duplex or multiplex PCR data obtained is comparable with that obtained from a singleplex PCR. This saves time, money, and reduces the amount of sample material needed. The special master mix in the QIAcuity Probe PCR Kit enables accurate quantification of up to 5 targets having widely differing abundance in a well of the QIAcuity Nanoplate. Probe-based detection from single to 5-plex with the QIAcuity Probe PCR Kit The unique combination of QIAGEN's proprietary and well-proven buffer technology optimized for the Nanoplate microfluidic along with the new QuantiNova DNA Polymerase delivers highly consistent results in terms of sensitivity, reproducibility and efficiency. botulinum group III, have rarely occurred, this new toxin type might pose little threat to human health.The QIAcuity Master Mixes for hydrolysis probe-based dPCR use the latest versions of QIAGEN’s high-quality DNA Polymerase. However, given that human infections with a similar toxin type, C. The risk for human infection with this new toxin type should also be investigated in future research. botulinum carrying the bont gene reported in this study. Further investigation is needed to determine the proportion of C. It should be noted that, because not all type C strains were subjected to sequencing, the presence of the bont gene (LC759602) as type C, as determined by typing PCR, might already exist in other samples. The bont gene (LC759602) was determined to be BoNT/C using PCR, which can easily distinguish between types C, D, CD, and DC of C. However, BoNT/DC has been reported to interact with gangliosides and protein receptors (synaptotagmin I and II) ( 5). Unlike other BoNTs, BoNT/C interacts only with gangliosides, and no protein receptor for this toxin has been identified ( 4). The bont gene (LC759602) possesses the bont/DC gene in the H C domain, suggesting that human susceptibility to this gene might differ from that of the reference BoNT/C toxin. The H C domain is involved in neurotoxin binding to specific receptors in peripheral nerve terminals. The bont gene (LC759602) has not been previously reported, and we propose its designation as a new subtype of C. Detailed analysis revealed that the bont gene (LC759602) had the bont/C gene or the bont/CD gene in the protease domain (LC) and the translocation domain (H N), as well as the bont/DC gene in the receptor-binding domain (H C) ( Table Figure). In comparison with other known bont genes, the bont gene of the strain sequenced in this study had the highest amino acid sequence similarity with the bont/C gene (90%) but was partially different from the reference bont/C gene ( Table). The obtained contig was assembled from reads of 59× coverage and 29 kbp in size. We analyzed sequencing data using CLC Genomics Workbench 22.0.2 (QIAGEN). We prepared genome-sequencing libraries using the QIAseq FX DNA Library Kit (QIAGEN) and sequenced the samples on the Illumina iSeq 100 ( ). botulinum with 20 mg/mL lysozyme in 20 mM Tris-HCl, 2 mM EDTA, and 1% Triton X-100 (pH 8.0), we extracted DNA using a QIAamp DNA Mini Kit (QIAGEN, ). The next-generation sequencing method was as follows: after treating C. Next-generation sequencing data revealed full-length coding regions of the bont gene of the isolated strains (GenBank accession no. We neutralized the toxin present in the specimens with type C botulinum antitoxin serum, and the isolated strain was found to carry the bont/C gene using PCR targeting the bont genes ( 3). A commercially prepared chicken dish was suspected to be the cause, but because no food was remaining, we were unable to conduct tests on it. botulinum were detected in 3 of the 4 specimens. A meal eaten in a domestic residence was the assumed cause, and 4 patients were affected. In 2021, foodborne botulism occurred in Kumamoto, Japan. Schematic diagrams of each functional domain between BoNT (C, CD, DC, D, and LC759602) in study of novel type C botulism strain in household outbreak, Japan.
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