An open-source, automated, and cost-effective platform for COVID-19 diagnosis and rapid portable genomic surveillance using nanopore sequencing.

The COVID-19 pandemic, caused by SARS-CoV-2, has emphasized the necessity for scalable diagnostic workflows using locally produced reagents and basic laboratory equipment with minimal dependence on global supply chains. We introduce an open-source automated platform for high-throughput RNA extraction and pathogen diagnosis, which uses reagents almost entirely produced in-house. This platform integrates our methods for self-manufacturing magnetic nanoparticles and qRT-PCR reagents-both of which have received regulatory approval for clinical use-with an in-house, open-source robotic extraction protocol. It also incorporates our "Nanopore Sequencing of Isothermal Rapid Viral Amplification for Near Real-time Analysis" (NIRVANA) technology, designed for tracking SARS-CoV-2 mutations and variants. The platform exhibits high reproducibility and consistency without cross-contamination, and its limit of detection, sensitivity, and specificity are comparable to commercial assays. Automated NIRVANA effectively identifies circulating SARS-CoV-2 variants. Our in-house, cost-effective reagents, automated diagnostic workflows, and portable genomic surveillance strategies provide a scalable and rapid solution for COVID-19 diagnosis and variant tracking, essential for current and future pandemic responses.

One Health Interactions of Chagas Disease Vectors, Canid Hosts, and Human Residents along the Texas-Mexico Border.

BACKGROUND: Chagas disease (Trypanosoma cruzi infection) is the leading cause of non-ischemic dilated cardiomyopathy in Latin America. Texas, particularly the southern region, has compounding factors that could contribute to T. cruzi transmission; however, epidemiologic studies are lacking. The aim of this study was to ascertain the prevalence of T. cruzi in three different mammalian species (coyotes, stray domestic dogs, and humans) and vectors (Triatoma species) to understand the burden of Chagas disease among sylvatic, peridomestic, and domestic cycles. METHODOLOGY/PRINCIPAL FINDINGS: To determine prevalence of infection, we tested sera from coyotes, stray domestic dogs housed in public shelters, and residents participating in related research studies and found 8%, 3.8%, and 0.36% positive for T. cruzi, respectively. PCR was used to determine the prevalence of T. cruzi DNA in vectors collected in peridomestic locations in the region, with 56.5% testing positive for the parasite, further confirming risk of transmission in the region. CONCLUSIONS/SIGNIFICANCE: Our findings contribute to the growing body of evidence for autochthonous Chagas disease transmission in south Texas. Considering this region has a population of 1.3 million, and up to 30% of T. cruzi infected individuals developing severe cardiac disease, it is imperative that we identify high risk groups for surveillance and treatment purposes.

The Biology of the Triatomine Bugs Native to South Central Texas and Assessment of the Risk They Pose for Autochthonous Chagas Disease Exposure.

Triatomine bugs are a group of hematophagous arthropods that can serve as biological vectors for Trypanosoma cruzi , the etiological agent of American trypanosomiasis (Chagas disease). Because of differences in the biology and feeding habits among triatomine species, some are more likely than others to be involved in zoonotic and/or human-to-human transmission cycles of T. cruzi . In an attempt to assess the risk for Chagas disease exposure in south-central Texas, human habitations across Texas Health Service Region 8 (HSR 8) and surrounding counties were surveyed for triatomines to characterize the geographic distribution, species-specific biology, and T. cruzi -infection prevalence better. Between May 2010 and August 2013, a total of 545 triatomines representing all 5 known indigenous species (Triatoma gerstaeckeri, Triatoma indictiva, Triatoma lecticularia, Triatoma sanguisuga, and Triatoma protracta woodi) were collected from 59 sites across the region. Triatoma gerstaeckeri was the species most commonly found in domestic and peridomestic ecotopes across Texas HSR 8, representing over 80% of the triatomines collected. Adult T. gerstaeckeri exhibited a seasonal dispersal pattern that began in late April, peaked in mid-May, and then continued into August. On homes with available crevices in the exterior walls, adult T. gerstaeckeri were observed emerging from or entering these protective microhabitats, suggesting possible opportunistic colonization of some exterior walls compartments. Laboratory testing of triatomine hindgut contents for T. cruzi by PCR demonstrated the adult T. gerstaeckeri-infection prevalence across Texas HSR 8 to be 64%. Monitoring peridomestic adult T. gerstaeckeri over the seasonal dispersal peak demonstrated statistically significant increases in both their T. cruzi -infection prevalence (P < 0.01) and tendency to invade human dwellings (P < 0.01) in the later aspect of the emergence peak. In addition to the adult insects, variably sized and staged nymphs were recovered from the inside of 6 separate homes across Texas HSR 8. The results of this study show that T. gerstaeckeri is a widespread and common triatomine species across Texas HSR 8 and documented it to have some notable synanthropic tendencies. The high prevalence of T. cruzi infection in native triatomines, and the high frequency with which T. gerstaeckeri is recovered from human habitations, suggests that there is a risk for human exposure to T. cruzi in Texas HSR 8. Because of this, Chagas disease should be considered on the list of differential diagnoses for cases of cardiac arrhythmia, dilated cardiomyopathy, or heart failure in south-central Texas.