Shigellais a leading cause of moderate-to-severe diarrhea globally and the causative agent of shigellosis and bacillary dysentery. Associated with 80 to 165 million cases of diarrhea and >13% of diarrheal deaths, in many regions,Shigellaexposure is ubiquitous while infection is heterogenous.

Background: Store-and-forward (S&F) telemedicine is thought to be most applicable in humanitarian settings. Unlike other kinds of telemental health (TMH), S&F requires engagement and active participation from healthcare providers in submitting text or audio-video clinical material for consultations. To implement such consultative systems there is a need to gauge providers' attitudes towards this technology. Materials and Methods: An electronic survey was sent to Syrian healthcare providers (physicians and nonphysicians) who are affiliated with humanitarian nongovernmental organizations managing Syrians affected by war. After a description of what TMH and S&F are, participants were asked about their attitudes towards such services. Results: Fifty-two providers responded to the electronic survey. Only results from providers inside Syria (n=30) are presented. The majority of respondents had no experience with TMH. Half of the providers believed that mental healthcare can be provided through S&F and that there would be a benefit from such services. Respondents reported that cultural (68%), financial (84%), and technical (80%) barriers do exist for such services. When asked, providers believed that patients would agree to be audiotaped (58%) for the purpose of S&F compared with being videotaped (15%) (p=0.007). Conclusions: Electronic surveys of healthcare workers in humanitarian settings are feasible. Providers in the Syrian humanitarian setting have little experience in TMH; however, they are open to using an S&F service while acknowledging cultural, financial, and technical barriers to the implementation.

Routinely used metagenomic next-generation sequencing (mNGS) techniques often fail to detect low-level viremia (<10⁴ copies/mL) and appear biased towards viruses with linear genomes. These limitations hinder the capacity to comprehensively characterize viral infections, such as those attributed to the Anelloviridae family. These near ubiquitous non-pathogenic components of the human virome have circular single-stranded DNA genomes that vary in size from 2.0 to 3.9 kb and exhibit high genetic diversity. Hence, species identification using short reads can be challenging. Here, we introduce a rolling circle amplification (RCA)-based metagenomic sequencing protocol tailored for circular single-stranded DNA genomes, utilizing the long-read Oxford Nanopore platform. The approach was assessed by sequencing anelloviruses in plasma drawn from people who inject drugs (PWID) in two geographically distinct cohorts. We detail the methodological adjustments implemented to overcome difficulties inherent in sequencing circular genomes and describe a computational pipeline focused on anellovirus detection. We assessed our protocol across various sample dilutions and successfully differentiated anellovirus sequences in conditions simulating mixed infections. This method provides a robust framework for the comprehensive characterization of circular viruses within the human virome using the Oxford Nanopore.

Nearly 300 million individuals live with chronic hepatitis B virus (HBV) infection (CHB), for which no curative therapy is available. As viral diversity is associated with pathogenesis and immunological control of infection, improved methods to characterize this diversity could aid drug development efforts. Conventionally, viral sequencing data are mapped/aligned to a reference genome, and only the aligned sequences are retained for analysis. Thus, reference selection is critical, yet selecting the most representative reference a priori remains difficult. We investigate an alternative pangenome approach which can combine multiple reference sequences into a graph which can be used during alignment. Using simulated short-read sequencing data generated from publicly available HBV genomes and real sequencing data from an individual living with CHB, we demonstrate alignment to a phylogenetically representative ‘genome graph’ can improve alignment, avoid issues of reference ambiguity, and facilitate the construction of sample-specific consensus sequences more genetically similar to the individual’s infection. Graph-based methods can, therefore, improve efforts to characterize the genetics of viral pathogens, including HBV, and have broader implications in host-pathogen research.