A case of viral meningitis due to Varicella Zoster virus infection in a young adult male.

A 36-year-old male with headaches was empirically treated for herpes simplex virus meningitis; CSF PCR testing later confirmed varicella zoster virus meningitis. Valacyclovir was increased to 2 g QID for remaining duration of therapy with full recovery. This case highlights the importance of comprehensive testing and proper treatment adjustment for rarer etiologies of aseptic meningitis.

Proposed Treatment Algorithm for Invasive Fungal Infections in Trauma Patients.

Background: Invasive fungal infections, most commonly caused by Mucorales species, are an underrecognized sequalae of traumatic injury that can complicate management of patients. The injury mechanism can introduce environmental spores into areas of the body normally not exposed to pathogens and this inoculation can progress rapidly to severe disease. The objective of this study was to present a case series of four trauma patients with invasive fungal infections that was used to develop an algorithm for work-up and treatment of these complex patients in future admissions. Patients and Methods: Four trauma patients who developed mucormycosis from two different hospitals are presented. One patient succumbed to their injuries whereas three were able to clear their infection with medical and surgical intervention. The surviving patients all had an infection of their lower extremity whereas the deceased patient had more extensive disease involving the thorax. Conclusions: Mucormycosis is a rare but significant post-trauma complication with substantial morbidity and mortality. Surgeons should be aware of this complication and maintain a high clinical suspicion because afflicted patients may not match the traditional clinical picture of a mucormycosis-susceptible patient. Close coordination with a pathology service is required for confirmation of the diagnosis and timely intervention can prevent debilitating loss of tissue or death. Additionally, consideration should be given to newer treatment modalities for management such as local tissue irrigation with an antifungal agent.

Design, validation and evaluation of a SYBR green-based quantitative PCR array for comprehensive analysis of adenovirus type 5 transcriptional patterns.

The adenoviral genome encodes coordinately expressed early and late gene transcriptional units that specify a complex collection of extensively spliced overlapping mRNAs. These complexities confound the generation of compatible, validated and optimized qPCR assays that permit comprehensive evaluation of adenoviral transcription. We have developed and evaluated a compilation of qPCR assays that represent the majority of the human adenovirus 5 (hAdV5) genome and allow for absolute and relative quantification of transcriptional activity. A panel of specific adenovirus gene primer pairs was designed through computational modeling to be compatible under a single reaction condition, precisely amplify spliced transcript products within each gene class, and not result in cellular or viral RNA/DNA background amplification. Primer pairs and reaction conditions were optimized to generate a single amplification product that was specific for its target amplicon with minimal intra-assay variability. The specificity of target amplicons was confirmed by dissociation curve analysis, gel electrophoresis and sequencing. In all, thirty-two primer sets representing specific gene products, as well as, pan early and late gene regions were validated under identical amplification conditions, thereby enabling a comprehensive assessment of adenoviral transcription within a single plate array. In order to generate positive control templates and to facilitate absolute quantification of gene expression, all target amplicons were cloned to create gene target-specific standards. These plasmid amplicon controls demonstrated that the SYBR qPCR assays exhibited optimal amplification efficiencies with a high sensitivity of detection to less than 10 copies and a linear amplification across at least eight orders of magnitude. The effectiveness and utility of the comprehensive adenoviral transcriptional array was assessed by investigating the changes in Ad5Wt gene expression at 72 versus 24 h post infection. Predictably, overall gene expression was globally increased at 72 h post infection; however, levels of E2 and Late transcripts exhibited the greatest increased expression, reflecting their necessity at this time point for genomic replication and virion assembly. Taken together, these data demonstrate that the adenoviral qPCR transcriptional array is a modular, scalable, and cost-effective method to comprehensively and accurately assess hAdV5 gene transcription. This array is broadly applicable to facilitate: adenoviral vector development; assessment of cell complementation of knockout viruses; antiviral mechanism of action evaluation; next-generation sequencing data validation.