BACKGROUND The prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) has changed unevenly over time around the world. Although whole genome sequencing is the gold standard for virus characterisation, the discovery of alpha VOC causing spike gene target failure (SGTF) result, when tested using an reverse transcription real-time polymerase chain reaction (RT-qPCR) assay, has provided a simple tool for tracking the frequencies of variants. OBJECTIVES The aim of this study was to investigate if a multiplex RT-qPCR assay (BioM 4Plex VOC) could be used to detect SARS-CoV-2 and to perform a VOC screening test in a single reaction tube. Here, we present the multicentre study evaluating this assay. METHODS Twelve laboratories have participated in the multicentre study. The BioM 4Plex VOC was distributed to them with detailed instructions of how to perform the test. They were asked to test the BioM 4Plex VOC in parallel with their routine Commercial SARS-CoV-2 diagnostic assay. Additionally, they were requested to select SARS-CoV-2-positive samples with genome sequenced and lineage definition according to PANGO lineage classification. FINDINGS The BioM 4Plex VOC and commercial RT-PCR assay are equally effective in detecting SARS-CoV-2. Results revealed a specificity of 96.5-100% [95% confidence interval (CI)], a sensitivity of 99.8-100% (95% CI), and an accuracy of 99.8-100% (95% CI). A 99% concordance rate was found between results from the BioM 4Plex VOC and that from available genome sequencing data. MAIN CONCLUSIONS The BioM 4Plex VOC provides an effective solution to detect SARS-CoV-2 infections and screening for VOCs in a single reaction. It is a straightforward method to help us monitor the frequency and distribution of VOCs and develop strategies to better cope with the pandemics. SARSCoV2 SARSCoV SARS CoV (SARS-CoV-2 (VOCs world characterisation SGTF (SGTF result realtime real RTqPCR RT qPCR (RT-qPCR Plex SARS-CoV- tube Here Additionally SARSCoV2positive SARSCoVpositive positive classification RTPCR PCR SARSCoV2. 2. 96.5100% 965100 96.5 100% 96 5 100 96.5-100 95% 95 [95 CI, CI , (CI)] 99.8100% 998100 99.8 99 8 99.8-100 (95 CI) CI. . data pandemics (SARS-CoV- SARS-CoV 5100 96.5100 96510 965 96. 9 10 96.5-10 [9 (CI) 8100 99.8100 99810 998 99. 99.8-10 (9 (SARS-CoV 510 96.510 9651 1 96.5-1 [ (CI 810 99.810 9981 99.8-1 ( 51 96.51 96.5- 81 99.81 99.8-

ABSTRACT In response to the Zika epidemics in Brazil, the ZDC molecular assay (Bio-Manguinhos) was developed and registered at the Brazilian Regulatory Agency of Health Surveillance - ANVISA. The circulation of Zika (ZIKV) Dengue (DENV) and Chikungunya (CHIKV) viruses and their clinical similarities are challenges to correctly diagnose these viruses. The simultaneous detection of ZIKV, DENV and CHIKV is an important tool for diagnosis and surveillance. Here, we present the analytical and clinical performance evaluation of ZDC molecular assay (Bio-Manguinhos) at the public health laboratories three years after its registration at ANVISA. The clinical performance demonstrates the ZDC molecular assay (Bio-Manguinhos) has 100% sensitivity and 100% specificity to detect and discriminate ZIKV, CHIKV, and DENV from clinical plasma samples. The ZDC molecular assay (Bio-Manguinhos) results were highly reproducible and no cross-reactivity was seen during testing with a panel of other infectious agents. In conclusion, the ZDC molecular assay (Bio-Manguinhos) is an accurate and reliable tool to monitor Zika, dengue and chikungunya infections in countries like Brazil with simultaneous circulation of the three viruses.

Abstract Hepatitis C virus (HCV) infection is a worldwide health problem. Nowadays, direct-acting antiviral agents (DAAs) are the main treatment for HCV; however, the high level of virus variability leads to the development of resistance-associated variants (RAVs). Thus, assessing RAVs in infected patients is important for monitoring treatment efficacy. The aim of our study was to investigate the presence of naturally occurring resistance mutations in HCV NS3 and NS5 regions in treatment-naïve patients. Ninety-six anti-HCV positive serum samples from blood donors at the Center of Hematology and Hemotherapy of Santa Catarina State (HEMOSC) were collected retrospectively in 2013 and evaluated in this study. HCV 1a (37.9%), 1b (25.3%), and 3a (36.8%) subtypes were found. The frequency of patients with RAVs in our study was 6.9%. The HCV NS5b sequencing reveled 1 sample with L320F mutation and 4 samples with the C316N/R polymorphism. The analysis of the NS3 region revealed the D168A/G/T (3.45%), S122G (1.15%), and V55A (2.3%) mutations. All samples from genotype 3a (36.8%) presented the V170 I/V non-synonymous mutation. In conclusion, we have shown that mutations in NS3 and NS5b genes are present in Brazilian isolates from therapy-naïve HCV patients.

Resumo A língua pilosa é uma patologia benigna, caracterizada clinicamente por placas hiperqueratinizadas na face dorsal da língua, semelhante a pelos, cuja coloração varia de despigmentada, esbranquiçada, amarelada, verde, acastanhada a preta. O diagnóstico é clínico, e em casos de placas esbranquiçadas, pode ser difícil diferenciar entre leucoplasia pilosa oral, leucoplasia potencialmente maligna ou carcinoma de células escamosas. Assim, o exame complementar de fluorescência óptica de campo amplo pode permitir uma melhor visualização do padrão local de hiperqueratinização semelhante à pelos, os quais são característicos de língua pilosa, facilitando o diagnóstico. Neste trabalho, um paciente do sexo masculino, 57 anos, foi encaminhado ao Departamento de Odontologia da Secretaria de Saúde de Divinópolis (Minas Gerais) por um clínico geral, visando o diagnóstico de uma possível lesão maligna na face dorsal da língua. O exame complementar por fluorescência óptica de campo amplo foi realizado. Para isso, foi empregado um dispositivo com diodo emissor de luz de alta potência, com luz centrada em um comprimento de onda de (400±10) nm e irradiância máxima de (0,04 ±0,008) W/cm2 para visualização de fluorescência. As imagens de fluorescência mostraram projeções de aparência semelhante à pelos na superfície dorsal da língua, sem aspectos de malignidade. A aparência similar à pelos é a principal característica da língua pilosa. Dessa maneira, o diagnóstico final foi estabelecido. Em conclusão, neste caso, o uso da fluorescência óptica de campo amplo permitiu um diagnóstico diferencial, sem a necessidade de uma biópsia incisional.

Abstract Hairy tongue is a benign pathology, characterized clinically by hyperkeratinized plaques on the dorsal surface of the tongue, hairlike, whose coloration ranges from unpigment, whitish, yellowish, green, brown to black. Diagnosis is clinical, and, in cases of whitish plaques, it may be difficult to differentiate between oral hairy leukoplakia, potentially malignant leukoplakia or squamous cell carcinoma. Thus, widefield optical fluorescence complementary examination may allow a better visualization of the local hairlike pattern of hyperkeratinization, typical of the hairy tongue, facilitating the diagnosis. In this work, a 57-year-old man was referred to the Dental Specialties Department of the Divinópolis Health Department (MG, Brazil) by a general dental practitioner, aiming a differential diagnosis of possible malignant lesion on the dorsal tongue surface. The complementary examination by wide-field optical fluorescence was performed. For this, it was employed a device with high-power light-emitting diode emitting light centered at a wavelength of (400±10) nm and maximum irradiance of (0.040±0.008) W/cm2 was used for fluorescence visualization. Fluorescence images showed projections of hairlike appearance in tongue dorsal surface with no aspects of malignancy. Hairlike appearance is the principal feature of hairy tongue. In this way, the final diagnosis was established. In conclusion, in this case, the use of widefield optical fluorescence in oral diagnostic routine provided a differential diagnosis, with no need of an incisional biopsy.

RESUMO O vitiligo é a doença crônica adquirida despigmentante mais comum da pele e/ou da mucosa. Entretanto, o vitiligo do tipo não classificado e subtipo de mucosa afetando apenas uma área da mucosa é considerado bastante incomum. O diagnóstico de vitiligo, independentemente do seu tipo, é clínico. No entanto, o uso de um dispositivo que permite a visualização da fluorescência tecidual pode ser útil para a confirmação do diagnóstico de vitiligo. Apresentamos o uso do dispositivo de exame complementar de fluorescência óptica de campo amplo para o diagnóstico de um caso incomum de vitiligo de mucosa localizado apenas em ângulos da boca.

SUMMARY Vitiligo is the most common depigmenting, chronic acquired disease of the skin and mucosa. However, vitiligo of an unclassified type and mucosal subtype affecting only one area of the mucosa is considered quite uncommon. The diagnosis of vitiligo, regardless of its type, is clinical. Nonetheless, a device that allows the visualization of the tissue fluorescence may be useful for confirming the diagnosis. We present the use of wide-field optical fluorescence device for complementary examination and diagnosis of unusual cases of mucosal vitiligo located only in angles of the mouth.

RESUMO O papiloma escamoso oral é um tumor benigno, cuja patogênese tem sido associada à infecção pelo papilomavírus humano. A infecção pelo papilomavírus humano é um dos fatores de risco associado ao desenvolvimento dos carcinomas cervicais, anogenitais, faríngeos, laríngeos e da cavidade oral. O papiloma escamoso oral pode acometer qualquer região da cavidade oral, e a transmissão do papilomavírus humano ocorre por contato direto, relação sexual ou de mãe para filho durante o parto. O diagnóstico é clínico e histopatológico, e a remoção cirúrgica representa o tratamento de escolha. Recentemente, a fluorescência óptica de campo amplo tem sido empregada como exame complementar ao exame clínico convencional, no rastreio de lesões patológicas orais e para delimitação de margens cirúrgicas. Relatamos um caso de papiloma escamoso oral com suas características clínicas e histopatológicas, sob a perspectiva da fluorescência óptica de campo amplo.

ABSTRACT Oral squamous papilloma is a benign tumor whose pathogenesis has been associated with human papillomavirus infection. Thus, it is noteworthy that human papillomavirus infection is one of the risk factors associated with the development of cervical, anogenital, pharynx, larynx and oral cavity carcinomas. Oral squamous papilloma can affect any region of the oral cavity, and transmission of human papillomavirus can occur by direct contact, sexual intercourse or from mother to child during delivery. The diagnosis is clinical and histopathological, with surgical removal representing the treatment of choice. Recently, widefield optical fluorescence has been used as a complementary examination to the conventional clinical examination in the screening of oral pathological lesions and for the delimitation of surgical margins. We report a case of oral squamous papilloma with its clinical, histopathological features and, in addition, from the perspective of wide field optical fluorescence.

ABSTRACT The Brazilian Public Health Service provides freely αPEG-IFN to treat patients infected with HCV. The primary goal of HCV therapy is the long-term elimination of HCV from the blood to reduce the risk of HCV associated complications and death. Patient viremia affects the treatment duration and response, thus influencing clinical decisions. We developed a high-throughput method to perform the quantification of RNA hepatitis C virus (HCV) virus load in plasma samples to monitor patients under treatment. The method is based on a duplex detection, in a one-step real-time RT-PCR assay and it has been validated according to the rules established by the official Brazilian regulatory agency (ANVISA). This new method was compared to a commercial kit (Cobas/Taqman HCV Test v2.0 - Roche), showing virus load results with significant correlation between them (p= 0,012) using commercial and clinical panels. In addition, 611 samples from patients treated with peguilated alfa-interferon (αPEG-IFN) from different regions of Brazil were analyzed. Our one-step real-time RT-PCR assay demonstrated good performance in viral load measurement and in treatment course monitoring, with acceptable sensitivity and specificity values.

OBJECTIVE: To assess the risk factors, incidence and severity of adverse drug reactions in in-patients. METHODS: This prospective study evaluated 472 patients treated at a teaching hospital in Brazil between 2010 and 2013 by five medical specialties: Internal Medicine, General Surgery, Geriatrics, Neurology, and Clinical Immunology and Allergy. The following variables were assessed: patient age, gender, comorbidities, family history of hypersensitivity, personal and family history of atopy, number of prescribed drugs before and during hospitalization, hospital diagnoses, days of hospitalization. The patients were visited every other day, and medical records were reviewed by the investigators to detect adverse drug reactions. RESULTS: There were a total of 94 adverse drug reactions in 75 patients. Most reactions were predictable and of moderate severity. The incidence of adverse drug reactions was 16.2%, and the incidence varied, according to the medical specialty; it was higher in Internal Medicine (30%). Antibiotics were the most commonly involved medication. Chronic renal failure, longer hospital stay, greater number of diagnoses and greater number of medications upon admission were risk factors. For each medication introduced during hospitalization, there was a 10% increase in the rate of adverse drug reaction. In the present study, the probability of observing an adverse drug reaction was 1 in 104 patients per day. CONCLUSIONS: Adverse drug reactions are frequent and potentially serious and should be better monitored in patients with chronic renal failure or prolonged hospitalization and especially in those on ‘polypharmacy’ regimens. The rational use of medications plays an important role in preventing adverse drug reactions.

OBJECTIVE: To assess the incidence of intra-operative immediate hypersensitivity reactions and anaphylaxis. METHODS: A cross-sectional observational study was conducted at the Department of Anesthesiology, University of São Paulo School of Medicine, Hospital das Clínicas, São Paulo, Brazil, from January to December 2010. We developed a specific questionnaire to be completed by anesthesiologists. This tool included questions about hypersensitivity reactions during anesthesia and provided treatments. We included patients with clinical signs compatible with immediate hypersensitivity reactions. Hhypersensitivity reactions were categorized according to severity (grades I-V). American Society of Anesthesiologists physical status classification (ASA 1-6) was analyzed and associated with the severity of hypersensitivity reactions. RESULTS: In 2010, 21,464 surgeries were performed under general anesthesia. Anesthesiologists answered questionnaires on 5,414 procedures (25.2%). Sixty cases of intra-operative hypersensitivity reactions were reported. The majority patients (45, 75%) had hypersensitivity reactions grade I reactions (incidence of 27.9:10,000). Fifteen patients (25%) had grade II, III or IV reactions (intra-operative anaphylaxis) (incidence of 7:10,000). No patients had grade V reactions. Thirty patients (50%) were classified as ASA 1. The frequency of cardiovascular shock was higher in patients classified as ASA 3 than in patients classified as ASA 1 or ASA 2. Epinephrine was administered in 20% of patients with grade III hypersensitivity reactions and in 50% of patients with grade II hypersensitivity reactions. CONCLUSIONS: The majority of patients had hypersensitivity reactions grade I reactions; however, the incidence of intra-operative anaphylaxis was higher than that previously reported in the literature. Patients with ASA 3 had more severe anaphylaxis; however, the use of epinephrine was not prescribed in all of these cases. Allergists and anesthesiologists should implement preventive measures to reduce the occurrence of anaphylaxis.