Early recognition of filoviral infections: Evaluation and development of diagnostic assays.

摘要

Filoviruses are responsible for sporadic, highly fatal outbreaks of hemorrhagic fever in human and non-human primates. Because of the mode of transmission, high fatality rates, and potential for spread throughout the community, early diagnosis of filovirus infections is crucial. Rapid, sensitive, and specific laboratory diagnostic tests are necessary to confirm outbreaks of filoviral hemorrhagic fever and to distinguish them from other diseases that can present with similar clinical syndromes.; Agent specific fluorogenic 5′ nuclease assays that are capable of detecting filovirus infections were developed and evaluated using the ABI PRISM 7700 Sequence Detection System™ and TaqMan ® chemistry. More specifically, a fluorogenic 5′ nuclease assay was developed and evaluated that is sensitive, specific, and capable of identifying all known strains of Marburg virus. In addition, a fluorogenic probe-based, one-tube RT-PCR assay that is capable of simultaneously identifying and differentiating between Ebola virus (Zaire strain) and Ebola virus (Sudan strain) was developed and evaluated. Both assays are compatible with emerging rapid nucleic acid analysis platforms.; Because agent-specific assays may not detect viral infection until after the onset of clinical signs, new diagnostic markers indicative of filoviral infection are needed. In order to identify new diagnostic markers, host response to filovirus infections was studied following infection of an in vitro culture of non-human primate alveolar macrophages with Ebola virus (Zaire strain). Fluorogenic 5′ nuclease assays specific for non-human primate cytokine and chemokine genes were used to characterize patterns of cytokine/chemokine mRNA expression following infection with Ebola virus (Zaire strain).