Cargando…

Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae

Sample preparation is a leading bottleneck in rapid detection of pathogenic bacteria. Here, we use Lyse-It(®) for bacterial cellular lysis, genomic DNA fragmentation, and protein release and degradation for both Listeria monocytogenes and Vibrio cholerae. The concept of Lyse-It(®) employs a conventi...

Descripción completa

Detalles Bibliográficos
Autores principales: Santaus, Tonya M., Li, Shan, Ladd, Paula, Harvey, Amanda, Cole, Shannon, Stine, O. Colin, Geddes, Chris D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6059484/
https://www.ncbi.nlm.nih.gov/pubmed/30044836
http://dx.doi.org/10.1371/journal.pone.0201070
Descripción
Sumario:Sample preparation is a leading bottleneck in rapid detection of pathogenic bacteria. Here, we use Lyse-It(®) for bacterial cellular lysis, genomic DNA fragmentation, and protein release and degradation for both Listeria monocytogenes and Vibrio cholerae. The concept of Lyse-It(®) employs a conventional microwave and Lyse-It(®) slides for intensely focused microwave irradiation onto the sample. High microwave power and a <60 second irradiation time allow for rapid cellular lysis and subsequent intracellular component release. The pathogenic bacteria are identified by quantitative polymerase chain reaction (qPCR), which subsequently demonstrates the viability of DNA for amplification post microwave-induced lysis. Intracellular component release, degradation, and detection of L. monocytogenes and V. cholerae has been performed and shown in this paper. These results demonstrate a rapid, low-cost, and efficient way for bacterial sample preparation on both food and water-borne Gram-positive and -negative organisms alike.