1,474 research outputs found
Log-normal model for microbial survival in heat sterilization
Log-normal model for microbial survival in heat sterilizatio
Multiplex STR amplification sensitivity in a silicon microchip
The demand for solutions to perform forensic DNA profiling outside of centralized laboratories is increasing. We here demonstrate highly sensitive STR amplification using a silicon micro-PCR (mu PCR) chip. Exploiting industry-standard semiconductor manufacturing processes, a device was fabricated that features a small form factor thanks to an integrated heating element covering three parallel micro-reactors with a reaction volume of 0.5 mu l each. Diluted reference DNA samples (1 ng-31 pg) were amplified on the mu PCR chip using the forensically validated AmpFISTR Identifier Plus kit, followed by conventional capillary electrophoresis. Complete STR profiles were generated with input DNA quantities down to 62 pg. Occasional allelic dropouts were observed from 31 pg downward. On-chip STR profiles were compared with those of identical samples amplified using a conventional thermal cycler for direct comparison of amplification sensitivity in a forensic setting. The observed sensitivity was in line with kit specifications for both mu PCR and conventional PCR. Finally, a rapid amplification protocol was developed. Complete STR profiles could be generated in less than 17 minutes from as little as 125 pg template DNA. Together, our results are an important step towards the development of commercial, mass-produced, relatively cheap, handheld devices for on-site testing in forensic DNA analysis
Silicon µPCR chip for forensic STR profiling with hybeacon probe melting curves
The demand to perform forensic DNA profiling outside of centralized laboratories and on the crime scene is increasing. Several criminal investigations would benefit tremendously from having DNA based information available in the first hours rather than days or weeks. However, due to the complexity and time-consuming nature of standard DNA fingerprinting methods, rapid and automated analyses are hard to achieve. We here demonstrate the implementation of an alternative DNA fingerprinting method in a single microchip. By combining PCR amplification and HyBeacon melting assays in a silicon Lab-ona-chip (LoC), a significant step towards rapid on-site DNA fingerprinting is taken. The small form factor of a LoC reduces reagent consumption and increases portability. Additional miniaturization is achieved through an integrated heating element covering 24 parallel micro-reactors with a reaction volume of 0.14 mu l each. The high level of parallelization allows the simultaneous analysis of 4 short tandem repeat (STR) loci and the amelogenin gender marker commonly included in forensic DNA analysis. A reference and crime scene sample can be analyzed simultaneously for direct comparison. Importantly, by using industry-standard semiconductor manufacturing processes, mass manufacturability can be guaranteed. Following assay design and optimization, complete 5-loci profiles could be robustly generated onchip that are on par with those obtained using conventional benchtop real-time PCR thermal cyclers. Together, our results are an important step towards the development of commercial, mass-produced, portable devices for on-site testing in forensic DNA analysis
The BioGRID Interaction Database: 2011 update
The Biological General Repository for Interaction Datasets (BioGRID) is a public database that archives and disseminates genetic and protein
interaction data from model organisms and humans
(http://www.thebiogrid.org). BioGRID currently holds 347 966
interactions (170 162 genetic, 177 804 protein) curated from both
high-throughput data sets and individual focused studies, as derived
from over 23 000 publications in the primary literature. Complete
coverage of the entire literature is maintained for budding yeast
(Saccharomyces cerevisiae), fission yeast (Schizosaccharomyces pombe)
and thale cress (Arabidopsis thaliana), and efforts to expand curation
across multiple metazoan species are underway. The BioGRID houses 48
831 human protein interactions that have been curated from 10 247
publications. Current curation drives are focused on particular areas
of biology to enable insights into conserved networks and pathways that
are relevant to human health. The BioGRID 3.0 web interface contains
new search and display features that enable rapid queries across
multiple data types and sources. An automated Interaction Management
System (IMS) is used to prioritize, coordinate and track curation
across international sites and projects. BioGRID provides interaction
data to several model organism databases, resources such as Entrez-Gene
and other interaction meta-databases. The entire BioGRID 3.0 data
collection may be downloaded in multiple file formats, including PSI MI
XML. Source code for BioGRID 3.0 is freely available without any
restrictions
Yersinia pseudotuberculosis serotype O : 1 infection in a captive Seba's short tailed-fruit bat (Carollia perspicillata) colony in Switzerland
BackgroundBetween February and April 2016, a slight increase in mortality was observed in a colony consisting of 400 captive Seba's short-tailed bats (Carollia perspicillata). These animals cohabited with other nocturnal animal species in a dome of a private zoo in Switzerland.ResultsGross and histological analysis of two (14.3%) out of the 13 animals submitted for necropsy within this period revealed a necrosuppurative pneumonia, hepatitis, splenitis, enterocolitis, and endometritis, with abundant intralesional colonies of Gram-negative rods. Yersinia (Y.) pseudotuberculosis serotype O:1 and biotype 1 belonging to the sequence type ST90 was isolated from the affected organs in both animals. Following this diagnosis, 1/4 of the colony (99 animals) was culled and submitted for gross and histopathological analysis, and a bacterial culture selective for Yersinia spp. of lung, liver, and spleen was performed. From these 99 animals, one gravid female was tested and found to be positive for Y. pseudotuberculosis in the absence of clinical symptoms and histopathological lesions. PCR analysis of altogether three bacterial isolates for virulence factors revealed the presence of the ail gene, and one isolate was also positive for the virF and yadA plasmid genes.ConclusionsThese findings suggest that Carollia perspicillata are susceptible to lethal yersiniosis but do not represent a regular reservoir for Y. pseudotuberculosis. Culling of 1/4 of the population was sufficient to limit the spread of this infection among the colony. Moreover, no infections were detected in cohabitant nocturnal animals and caretakers, indicating that the zoonotic risk in this case was low.Peer reviewe
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