A Multiplex Nanopore Sequencing Approach for the Detection of Multiple Arboviral Species.

The emergence and continued geographic expansion of arboviruses and the growing number of infected people have highlighted the need to develop and improve multiplex methods for rapid and specific detection of pathogens. Sequencing technologies are promising tools that can help in the laboratory diagnosis of conditions that share common symptoms, such as pathologies caused by emerging arboviruses. In this study, we integrated nanopore sequencing and the advantages of reverse transcription polymerase chain reaction (RT-PCR) to develop a multiplex RT-PCR protocol for the detection of Chikungunya virus (CHIKV) and several orthoflaviviruses (such as dengue (Orthoflavivirus dengue), Zika (Orthoflavivirus zikaense), yellow fever (Orthoflavivirus flavi), and West Nile (Orthoflavivirus nilense) viruses) in a single reaction, which provides data for sequence-based differentiation of arbovirus lineages.

The impact of an enhanced health surveillance system for COVID-19 management in Serrana, Brazil.

Objective: To describe the successful implementation of an enhanced public health surveillance system based on early detection, tracing contacts, and patient follow-up and support. Study design: A prospective observational cohort study conducted in Serrana, São Paulo State, Brazil. Methods: The implementation was based on four axes: increasing the access to SARS-CoV-2 testing; correct swab collection; testing patients with mild symptoms; and patient follow-up. Positivity rate, patient demographic and clinical characteristics, dynamics of disease severity, SARS-CoV-2 genome evolution, and the impact on COVID-19 research were assessed from August 23, 2020 to February 6, 2021 (between epidemiological week 35/2020 and 5/2021, a total of 24 weeks). Results: The number of sites collecting rt-PCR for SARS-CoV-2 was increased from one to seven points and staff was trained in the correct use of personal protective equipment and in the swab collection technique. During the study period, 6728 samples were collected from 6155 participants vs. 2770 collections in a similar period before. SARS-CoV-2 RNA was detected in 1758 (26.1%) swabs vs. 1117 (36.7%) before the implementation of the surveillance system (p < 0.001). Positivity rates varied widely between epidemiological weeks 35/2020 and 5/2021 (IQR, 12.8%-31.3%). Out of COVID-19 patients, 91.1% were adults at a median age of 35 years (IQR, 25-50 years), 42.6% were men and 57.4% were women, with a SARS-CoV-2 positivity rate of 28.6% and 24.4% (p < 0.001), respectively. The most common symptoms were headache (72.6%), myalgia (65.0%), and cough (61.7%). Comorbidities were found in 20.8% of patients, the most common being hypertension and diabetes. According to the World Health Organization clinical progression scale, 93.5% of patients had mild disease, 1.6% were hospitalized with moderate disease, 3.2% were hospitalized with severe disease, and 1.4% died. The enhanced surveillance system led to the development of COVID-19 related research. Conclusions: The enhanced surveillance system in Serrana improved COVID-19 understanding and management. By integrating community and academic institutions, it was possible to monitor SARS-CoV-2 positive cases and variants, follow the epidemic trend, guide patients, and develop relevant research projects.

High-Dose Convalescent Plasma for Treatment of Severe COVID-19.

To assess whether high-dose coronavirus disease (COVID-19) convalescent plasma (CCP) transfusion may benefit patients with severe COVID-19, we conducted a multicenter randomized trial in Brazil. Patients with severe COVID-19 who were within 10 days of initial symptom onset were eligible. Patients in the CCP group received 3 daily doses of CCP (600 mL/d) in addition to standard treatment; control patients received standard treatment only. Primary outcomes were death rates at days 30 and 60 of study randomization. Secondary outcomes were ventilator-free days and hospital-free days. We enrolled 107 patients: 36 CCP and 71 control. At day 30, death rates were 22% for CCP and 25% for the control group; at day 60, rates were 31% for CCP and 35% for control. Needs for invasive mechanical ventilation and durations of hospital stay were similar between groups. We conclude that high-dose CCP transfused within 10 days of symptom onset provided no benefit for patients with severe COVID-19.

Genetic diversity and chemical variability of Lippia spp. (Verbenaceae).

BACKGROUND: The genus Lippia comprises 150 species, most of which have interesting medicinal properties. Lippia sidoides (syn. L. origanoides) exhibits strong antimicrobial activity and is included in the phytotherapy program implemented by the Brazilian Ministry of Health. Since species of Lippia are morphologically very similar, conventional taxonomic methods are sometimes insufficient for the unambiguous identification of plant material that is required for the production of certified phytomedicines. Therefore, genetic and chemical analysis with chemotype identification will contribute to a better characterization of Lippia species. METHODS: Amplified Length Polymorphism and Internal Transcribed Spacer molecular markers were applied to determine the plants' genetic variability, and the chemical variability of Lippia spp. was determined by essential oil composition. RESULTS: Amplified Length Polymorphism markers were efficient in demonstrating the intra and inter-specific genetic variability of the genus and in separating the species L. alba, L. lupulina and L. origanoides into distinct groups. Phylogenetic analysis using Amplified Length Polymorphism and markers produced similar results and confirmed that L. alba and L. lupulina shared a common ancestor that differ from L. origanoides. Carvacrol, endo-fenchol and thymol were the most relevant chemical descriptors. CONCLUSION: Based on the phylogenetic analysis it is proposed that L. grata should be grouped within L. origanoides due to its significant genetic similarity. Although Amplified Length Polymorphism and Internal Transcribed Spacer markers enabled the differentiation of individuals, the genotype selection for the production of certified phytomedicines must also consider the chemotype classification that reflects their real medicinal properties.