Beckermyers8452

Detection of HHV-6 by FA-ME led to discontinuation of acyclovir within 12.0 hours in all 12 patients empirically treated with acyclovir. Six of the twelve patients were started on ganciclovir therapy within 6.8 hours; four of which were treated specifically for HHV-6 infection, whereas therapy was discontinued in the remaining 2 patients. CSF parameters were not generally predictive of HHV-6 positivity. The presence of ciHHV-6 was confirmed in 3 of 18 patients able to be tested.Conclusions Five of the 25 patients included in the study were diagnosed with HHV-6 meningitis/meningoencephalitis. FA-ME results led to discontinuation of empirical anti-viral in 12 patients and appropriate initiation of ganciclovir in 4 patients. In our institution, detection of HHV-6 using FA-ME led to faster establishment of disease etiology and optimization of antimicrobial therapy. Copyright © 2020 American Society for Microbiology.Specific astrovirus genotype species are associated with neurological disease and encephalitis in humans, mink, pigs, sheep and cattle (1).…. Copyright © 2020 American Society for Microbiology.Coronavirus Disease 19 (COVID-19), has become the Public Health Emergency of International Concern.…. Copyright © 2020 American Society for Microbiology.Background Klebsiella species are problematic pathogens in neonatal units and may cause outbreaks, for which sources of transmission can be challenging to elucidate. We describe the use of whole genome sequencing (WGS) to investigate environmental sources of transmission during an outbreak of extended-spectrum-β-lactamase (ESBL)-producing Klebsiella michiganensis colonizing neonates.Methods Ceftriaxone-resistant Klebsiella spp. isolated from neonates (or their mothers) and the hospital environment were included. Short-read (Illumina) and long-read (MinION, Oxford Nanopore Technologies) sequencing was used to confirm species taxonomy, define antimicrobial resistance genes and determine phylogenetic relationships using single nucleotide polymorphism (SNP) profiling.Results A total of 21 organisms (10 patient-derived and 11 environmental isolates) were sequenced. Standard laboratory methods identified the outbreak strain as an ESBL-producing Klebsiella oxytoca, but taxonomic assignment from WGS data suggested closer identity to Klebsiella michiganensis. KY 12420 Strains isolated from multiple detergent dispensing bottles were either identical or closely related by SNP comparison. Detergent bottles contaminated by K. michiganensis had been used for washing milk-expressing equipment. No new cases were identified once the detergent bottles were removed.Conclusions Environmental reservoirs may be an important source in outbreaks of multi-drug resistant organisms. WGS, in conjunction with traditional epidemiological investigation, can be instrumental in revealing routes of transmission and guiding infection control responses. Copyright © 2020 American Society for Microbiology.Microscopy and rapid diagnostic tests (RDTs) are the main diagnostic tools for malaria but fail to detect low-density parasitemia that are important for maintaining malaria transmission. To complement existing diagnostic methods, an isothermal reverse transcription recombinase polymerase amplification and lateral flow assay (RT-RPA) was developed. We compared the performance with ultrasensitive reverse transcription qPCR (RT-qPCR) using nucleic acid extracts from blood (n=114) obtained after standardized controlled human malaria infection (CHMI) with Plasmodium falciparum sporozoites. As a preliminary investigation, we also sampled asymptomatic individuals (n=28) in a malaria endemic area (Lambaréné, Gabon) to validate RT-RPA and assess its performance with unprocessed blood samples (dbRT-RPA). In 114 samples analyzed from CHMI-trials, the positive percent agreement to uRT-qPCR was 90% (95% CI 80-96). The negative percent agreement was 100% (95% CI 92-100). The lower limit of detection was 64 parasites/ml. In Gabon, RT-RPA was 100% accurate with asymptomatic volunteers (n=28) while, simplified dbRT-RPA showed 89% accuracy. In a subgroup analysis, RT-RPA detected 9/10 of RT-qPCR positive samples, while loop-mediated isothermal amplification (LAMP) detected 2/10. RT-RPA is a reliable diagnostic test for asymptomatic low-density infections. It is particularly useful in settings where uRT-qPCR is difficult to implement. Copyright © 2020 American Society for Microbiology.Lyme disease (LD) is an increasing public health problem. Current laboratory testing is insensitive in early infection, the stage at which appropriate treatment is most effective in preventing disease sequela. The Lyme Disease Biobank (LDB) collects samples from individuals with symptoms consistent with early LD presenting with or without erythema migrans (EM) or an annular, expanding skin lesion, and uninfected individuals from endemic areas. Samples were collected from 550 participants (298 cases and 252 controls) according to IRB-approved protocols and shipped to a centralized biorepository. Testing was performed to confirm the presence of tick-borne pathogens by real-time PCR, and a subset of samples was tested for Borrelia burgdorferi by culture. Serology using the CDC's standard two-tiered testing algorithm (STTTA) for LD was performed on all samples. LD diagnosis was supported by laboratory testing in 82 cases, including positive STTTA, PCR, culture, or 2 positive ELISA's with EM >5 cm, while the remaining 216 cases had negative laboratory testing results. For the controls, 43 were positive on at least one of the tiers, and 6 were positive by STTTA. This collection highlights and reinforces the known limitations of serologic testing in early LD, with only 29% of individuals presenting with EM >5 cm yielding a positive result using the STTTA. Aliquots of whole blood, serum, and urine from clinically characterized patients with and without LD are available to investigators in academia and industry for evaluation or development of novel diagnostic assays for LD, to continue to improve upon currently available methods. Copyright © 2020 Horn et al.Introduction Serological testing for nasopharyngeal carcinoma (NPC) has recently been reinvigorated by the implementation of novel Epstein-Barr virus (EBV) specific IgA and IgG antibodies from a proteome array. Although proteome arrays are well-suited for comprehensive antigen selection, they are not applicable for large-scale studies.Methods We have adapted a 13-marker EBV antigen signature for NPC risk identified by proteome arrays to multiplex serology, to establish an assay for large-scale studies. Taiwanese NPC cases (n=175) and matched controls (n=175) were used for assay validation. Spearman's correlation was calculated and the diagnostic value of all multiplex markers was assessed independently using the area under the receiver operating characteristic curve (AUC). Two refined signatures were identified using stepwise logistic regression, and internally validated with 10-fold cross validation.Results Array and multiplex serology showed strong correlation for each individual EBV-marker, as well as for a 13-marker combined model on continuous data. Two refined signatures with either four (LF2 and BGLF2 IgG, LF2 and BMRF1 IgA) or two antibodies (LF2 and BGLF2 IgG) on dichotomous data were identified as the most parsimonious set of serological markers able to distinguish NPC cases from controls with AUCs of 0.992 (95% CI 0.983-1.000) and 0.984 (95% CI 0.971-0.997), respectively. link2 Neither differed significantly from the 13-marker model (AUC = 0.992; 95% CI 0.982-1.000). All models were internally validated.Conclusion Multiplex serology successfully validated the original EBV proteome microarray data. Two refined signatures of four and two antibodies were capable of detecting NPC with 99.2% and 98.4% accuracy. Copyright © 2020 American Society for Microbiology.The skin-colonizing commensal bacterium Staphylococcus epidermidis is a leading cause of hospital-acquired and device-related infections. Its pathogenicity in humans is largely due to its propensity to form biofilms, surface-adherent bacterial accumulations that are remarkably resistant to chemical and physical stresses. Accumulation-associated protein (Aap) from S. epidermidis has been shown to be necessary and sufficient for mature biofilm formation and catheter infection. Aap contains up to 17 tandem B-repeat domains, capable of zinc-dependent assembly into twisted, rope-like intercellular filaments in the biofilm. Using microscopic and biophysical techniques, we show here that longer Aap B-repeat constructs assemble further into zinc-dependent functional amyloid fibers. We observed such amyloid fibers by confocal microscopy during both early and late stages of S. epidermidis biofilm formation and we confirmed that extracellular fibrils from these biofilms contain Aap. Unlike what has been observed for amyloidogenic biofilm proteins from other bacteria, which typically use chaperones or initiator proteins to initiate amyloid assembly, our findings indicate that Aap from S. epidermidis requires Zn2+ as a catalyst that drives amyloid fiber formation, similar to many mammalian amyloid-forming proteins that require metals for assembly. This work provides detailed insights into S. epidermidis biofilm formation and architecture that improve our understanding of persistent staphylococcal infections. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.Animal cells use pattern-recognition receptors (PRRs) to detect specific pathogens. Pathogen detection mounts an appropriate immune response, including interferon and cytokine induction. The intracellular PRR-signaling pathways that detect DNA viruses have been characterized, particularly in myeloid cells. In these pathways, cGMP-AMP synthase (cGAS) and the pyrin and HIN domain family member (PYHIN) protein interferon g-inducible protein 16 (IFI16) detect DNA and signal via stimulator of interferon genes protein (STING). However, although airway epithelial cells are frontline sentinels in detecting pathogens, information on how they respond to DNA viruses is limited, and the roles of PYHIN proteins in these cells are unknown. Here, we examined expression and activities of cGAS, STING, and PYHINs in human lung epithelial cells. link3 A549 epithelial cells, commonly used for RNA-sensing studies, failed to respond to DNA because they lacked STING expression, and ectopic STING expression restored a cGAS-dependent DNA response in these cells. In contrast, NuLi-1 immortalized human bronchial epithelial cells did express STING, which was activated after DNA stimulation and mediated DNA-dependent gene induction. PYHIN1, which like IFI16 has been proposed to be a viral DNA sensor, was the only PYHIN protein expressed in both airway epithelial cell types. However, rather than having a role in DNA sensing, PYHIN1 induced proinflammatory cytokines in response to interleukin-1 (IL-1) or tumor necrosis factor α (TNF-α) stimulation. Of note, PYHIN1, via its HIN domain, directly induced IL-6 and TNF-α transcription, revealing that PYHIN proteins play a role in proinflammatory gene induction in airway epithelial cells. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.