RESUMEN
The revelation in May 2015 of the shipment of γ irradiation-inactivated wild-type Bacillus anthracis spore preparations containing a small number of live spores raised concern about the safety and security of these materials. The finding also raised doubts about the validity of the protocols and procedures used to prepare them. Such inactivated reference materials were used as positive controls in assays to detect suspected B. anthracis in samples because live agent cannot be shipped for use in field settings, in improvement of currently deployed detection methods or development of new methods, or for quality assurance and training activities. Hence, risk-mitigated B. anthracis strains are needed to fulfill these requirements. We constructed a genetically inactivated or attenuated strain containing relevant molecular assay targets and tested to compare assay performance using this strain to the historical data obtained using irradiation-inactivated virulent spores.
Asunto(s)
Carbunco/microbiología , Bacillus anthracis/fisiología , Bacillus anthracis/efectos de la radiación , Radiación , Esporas Bacterianas/efectos de la radiación , Animales , Bacillus anthracis/virología , Toxinas Bacterianas/genética , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Ratones , Mutagénesis Insercional , Plásmidos/genética , Recombinación Genética , Reproducibilidad de los Resultados , Virulencia , Secuenciación Completa del GenomaRESUMEN
Nanopore sequencing, a novel genomics technology, has potential applications for routine biosurveillance, clinical diagnosis, and outbreak investigation of virus infections. Using rapid sequencing of unamplified RNA/cDNA hybrids, we identified Venezuelan equine encephalitis virus and Ebola virus in 3 hours from sample receipt to data acquisition, demonstrating a fieldable technique for RNA virus characterization.