A Multi-Laboratory Evaluation of Commercial Monkeypox Virus Molecular Tests.

In this report, we describe the first national scale multi-laboratory evaluation of monkeypox virus (MPXV) DNA commercial PCR kits. The objective of this study was to evaluate 2 kits by different diagnostic laboratories across Israel. Ten standardized samples were tested simultaneously using the Novaplex (15 laboratories) and Bio-Speedy (seven laboratories) kits. An in-house assay based on previously published reactions was used as reference. Comparison of the results showed high intra-assay agreement between laboratories, with small variations for most samples. The in-house assay had an analytical detection limit of less than 10 copies per reaction. While the 2 commercial kits were able to detect specimens with low viral loads similarly to the in-house assay, significant differences were observed, in the Cq values and relative fluorescence (RF), between the assays. The RF signal of the in-house and Bio-Speedy assays ranged between 5,000 and 10,000 RFU, while the signal in the Novaplex assay was less than 600 RFU. Due to the kit measurement protocol, the Cq values of the Bio-Speedy kit were 5 to 7.5 cycles lower than those of the in-house assay. On the contrary, the Cq values of the Novaplex kit were significantly higher than those of the in-house assay, with differences of 3 to 5 cycles per sample. Our results suggest that while all assays were similar in their overall sensitivity, direct comparison of Cq values between them may be misleading. To our knowledge, this is the first methodical evaluation of commercial MPX test kits. We therefore anticipate that this study would help diagnostic laboratories in choosing a specific MPX detection assay. IMPORTANCE To the best of our knowledge, this study is the first methodical evaluation of commercial kits designed for Monkeypox virus detection. This was done by performing the same tests using the same sample set in multiple laboratories, simultaneously, on a national scale. It therefore provides important and unique information on the performance of such kits and provides a guideline for choosing the assay of choice for monkeypox virus diagnosis in a standard diagnostic laboratory. It also demonstrates potential complications when trying to compare the results of different assays, even when testing exactly the same samples, under identical conditions.

National Scale Real-Time Surveillance of SARS-CoV-2 Variants Dynamics by Wastewater Monitoring in Israel.

In this report, we describe a national-scale monitoring of the SARS-CoV-2 (SC-2) variant dynamics in Israel, using multiple-time sampling of 13 wastewater treatment plants. We used a combination of inclusive and selective quantitative PCR assays that specifically identify variants A19/A20 or B.1.1.7 and tested each sample for the presence and relative viral RNA load of each variant. We show that between December 2020 and March 2021, a complete shift in the SC-2 variant circulation was observed, where the B.1.1.7 replaced the A19 in all examined test points. We further show that the normalized viral load (NVL) values and the average new cases per week reached a peak in January 2021 and then decreased gradually in almost all test points, in parallel with the progression of the national vaccination campaign, during February-March 2021. This study demonstrates the importance of monitoring SC-2 variant by using a combination of inclusive and selective PCR tests on a national scale through wastewater sampling, which is far more amendable for high-throughput monitoring compared with sequencing. This approach may be useful for real-time dynamics surveillance of current and future variants, such as the Omicron (BA.1, BA.2) and other variants.

Monitoring of Enterovirus D68 Outbreak in Israel by a Parallel Clinical and Wastewater Based Surveillance.

Enterovirus D68 (EVD68) was recently identified as an important cause of respiratory illness and acute flaccid myelitis (AFM), mostly in children. Here, we examined 472 pediatric patients diagnosed with severe respiratory illness and screened for EVD68 between April and October 2021. In parallel, samples collected from a wastewater treatment plant (WWTP) covering the residential area of the hospitalized patients were also tested for EVD68. Of the 472 clinical samples evaluated, 33 (7%) patients were positive for EVD68 RNA. All wastewater samples were positive for EVD68, with varying viral genome copy loads. Calculated EVD68 genome copies increased from the end of May until July 2021 and dramatically decreased at the beginning of August. A similar trend was observed in both clinical and wastewater samples during the period tested. Sequence analysis of EVD68-positive samples indicated that all samples originated from the same branch of subclade B3. This study is the first to use wastewater-based epidemiology (WBE) to monitor EVD68 dynamics by quantitative detection and shows a clear correlation with clinically diagnosed cases. These findings highlight the potential of WBE as an important tool for continuous surveillance of EVD68 and other enteroviruses.

Specific Detection of SARS-CoV-2 Variants B.1.1.7 (Alpha) and B.1.617.2 (Delta) Using a One-Step Quantitative PCR Assay.

In this report, we describe the development of a reverse transcription-quantitative PCR (RT-qPCR) assay, termed Alpha-Delta assay, which can detect all severe acute respiratory syndrome coronavirus 2 (SC-2) variants and distinguish between the Alpha (B.1.1.7) and Delta (B.1.617.2) variants. The Alpha- and Delta-specific reactions in the assay target mutations that are strongly linked to the target variant. The Alpha reaction targets the D3L substitution in the N gene, and the Delta reaction targets the spike gene 156 to 158 mutations. Additionally, we describe a second Delta-specific assay that we use as a confirmatory test for the Alpha-Delta assay that targets the 119 to 120 deletion in the Orf8 gene. Both reactions have similar sensitivities of 15 to 25 copies per reaction, similar to the sensitivity of commercial SC-2 detection tests. The Alpha-Delta assay and the Orf8119del assay were successfully used to classify clinical samples that were subsequently analyzed by whole-genome sequencing. Lastly, the capability of the Alpha-Delta assay and Orf8119del assay to identify correctly the presence of Delta RNA in wastewater samples was demonstrated. This study provides a rapid, sensitive, and cost-effective tool for detecting and classifying two worldwide dominant SC-2 variants. It also highlights the importance of a timely diagnostic response to the emergence of new SC-2 variants with significant consequences on global health. IMPORTANCE The new assays described herein enable rapid, straightforward, and cost-effective detection of severe acute respiratory syndrome coronavirus 2 (SC-2) with immediate classification of the examined sample as Alpha, Delta, non-Alpha, or non-Delta variant. This is highly important for two main reasons: (i) it provides the scientific and medical community with a novel diagnostic tool to rapidly detect and classify any SC-2 sample of interest as Alpha, Delta, or none and can be applied to both clinical and environmental samples, and (ii) it demonstrates how to respond to the emergence of new variants of concern by developing a variant-specific assay. Such assays should improve our preparedness and adjust the diagnostic capacity to serve clinical, epidemiological, and research needs.

Rapid and High-Throughput Reverse Transcriptase Quantitative PCR (RT-qPCR) Assay for Identification and Differentiation between SARS-CoV-2 Variants B.1.1.7 and B.1.351.

Emerging SARS-CoV-2 (SC-2) variants with increased infectivity and vaccine resistance are of major concern. Rapid identification of such variants is important for the public health decision making and to provide valuable data for epidemiological and policy decision making. We developed a multiplex reverse transcriptase quantitative PCR (RT-qPCR) assay that can specifically identify and differentiate between the emerging B.1.1.7 and B.1.351 SC-2 variants. In a single assay, we combined four reactions-one that detects SC-2 RNA independently of the strain, one that detects the D3L mutation, which is specific to variant B.1.1.7, one that detects the 242 to 244 deletion, which is specific to variant B.1.351, and the fourth reaction, which identifies the human RNAseP gene, serving as an endogenous control for RNA extraction integrity. We show that the strain-specific reactions target mutations that are strongly associated with the target variants and not with other major known variants. The assay's specificity was tested against a panel of respiratory pathogens (n = 16), showing high specificity toward SC-2 RNA. The assay's sensitivity was assessed using both in vitro transcribed RNA and clinical samples and was determined to be between 20 and 40 viral RNA copies per reaction. The assay performance was corroborated with Sanger and whole-genome sequencing, showing complete agreement with the sequencing results. The new assay is currently implemented in the routine diagnostic work at the Central Virology Laboratory, and may be used in other laboratories to facilitate the diagnosis of these major worldwide-circulating SC-2 variants. IMPORTANCE This study describes the design and utilization of a multiplex reverse transcriptase quantitative PCR (RT-qPCR) to identify SARS-COV-2 (SC2) RNA in general and, specifically, to detect whether it is of lineage B.1.1.7 or B.1.351. Implementation of this method in diagnostic and research laboratories worldwide may help the efforts to contain the COVID-19 pandemic. The method can be easily scaled up and be used in high-throughput laboratories, as well as small ones. In addition to immediate help in diagnostic efforts, this method may also help in epidemiological studies focused on the spread of emerging SC-2 lineages.

Formação do psicólogo para atuação comunitária no contexto das drogas: uma análise das matrizes curriculares / Training in community psychology action in the context of drugs: an analysis of curricular matrices

A psicologia comunitária enfatiza a prevenção e promoção da saúde. Neste contexto, questões relacionadas ao consumo de drogas emergem enquanto demanda de trabalho. Diante disso, este estudo teve como objetivo analisar a formação em psicologia com relação à atuação no contexto comunitário frente às demandas relacionadas ao uso de drogas, a partir da análise das matrizes curriculares. Para tanto, realizou-se uma análise das matrizes curriculares dos cursos de psicologia do Sul do país. Dos 102 cursos analisados, 21 oferecem disciplinas sobre drogas e 73 sobre comunidade. Em 12 instituições, foi identificada a presença de ambas as disciplinas. Tais resultados indicam que, embora a formação em psicologia tenha avançado quanto à psicologia comunitária, o tema “drogas” ainda é pouco explorado, o que dificulta ações pautadas num aporte teórico-metodológico que convirjam com evidências científicas relacionadas ao tema. É imprescindível uma formação que prepare o profissional para atuar de forma ética e comprometida com demandas relacionadas à questão do consumo de drogas presentes na comunidade.(AU)

Community psychology emphasizes prevention and health promotion. In this context, issues related to drug use emerge as a demand for work. Therefore, this study aimed to discuss the training of the psychologist as an ethical commitment to act in the community context regarding the demands related to drug use. For that, an analysis of the curricular matrices of the psychology courses of the country ́s South was carried out. Of the 102 courses analyzed, 21 offer courses on drugs and 73 courses on community. In 12 institutions, the presence of both disciplines was identified. Although the formation of the Psychologist has advanced in relation to community psychology, the theme "drugs" is still little explored, which hampers actions based on a theoretical and methodological contribution that converge with scientific evidence related to the theme. It is essential a training that prepares the professional to act with ethics and committed to the demands of the community.(AU)

Usuários de crack: histórias de vida e subjetividade em narrativas audiovisuais / Crack users: life stories and subjectivity in audiovisual narratives / Usuarios de crack: historias de vida y subjetividad en narrativas audiovisuales

Constituindo-se o uso problemático de crack como grave questão de Saúde Pública, este artigo é parte de dissertação que teve como objetivo compreender a relação entre histórias de vida e projetos de ser dos sujeitos usuários de crack com o consumo da substância e problemas a ele relacionados. Solicitou-se a cinco participantes que fizessem vídeos sobre suas histórias de vida, e que falassem sobre o processo de reflexão sobre suas trajetórias e aspectos de suas vidas presentes ou não nos vídeos. Os dados produzidos foram analisados conforme a Análise de Conteúdo proposta por Ruiz-Olabuénaga. Percebeu-se o ato de consumir crack como mais um dos atos humanos a constituir o sujeito e sua biografia, embora se torne problemático quando promove "suspensão" das outras possibilidades de vida. Verificou-se que eles tenderam a adotar discursos padronizados sobre drogas e usuários, ainda que contradigam suas próprias histórias de vida, embora reconheçam que há algo de particular na sua trajetória de uso e nos processos de tratamento/ recuperação. Esta pesquisa indica a necessidade de estratégias que fomentem a reflexão dos usuários sobre o sujeito por trás da droga, destacando aspectos universais e singulares de suas biografias, tendo na construção de narrativas autobiográficas audiovisuais um instrumento clinicamente potente.

Considering crack abuse as a problem of Public Health, this research aimed to understand the relationship between the life stories and the "being projects" of crack users with the consumption of the substance and related problems. 5 participants were asked to do videos about their life stories and to talk about the process of reflection on their trajectories, thinking about aspects addressed or not of their lives in the films produced by them. The data produced were analyzed according to the Content Analysis proposed by Ruiz-Olabuénaga. It was noticed the act of consuming crack as another human act who creates the subject and his biography, though it is problematic when promotes a "suspension" of the other possibilities of life. It was found that the users themselves tend to adopt standardized discourses on drugs and users, even when they contradict his own life experience, while recognizing that there is something particular in his phenomenon of the use and recovery and treatment’s processes. This research indicates that it is necessary to promote users' reflection about the "person" behind the drug, highlighting the universal and individual aspects of their biographies, with the construction of autobiographical audiovisual narratives as a clinically powerful tool.

Constituyéndose el abuso de crack como grave problema de salud pública, este artículo es parte de disertación que tuvo como objetivo comprender la relación entre las historias de vida y proyectos de ser de los consumidores de crack con el consumo de la sustancia y problemas relacionados. Se pidió a 5 participantes que hiciesen vídeos sobre sus historias de vida y hablasen sobre el proceso de reflexión sobre sus trayectorias y los aspectos de su vida presentes o no en los videos. Los datos obtenidos fueron analizados de acuerdo al análisis de contenido de Ruiz-Olabuénaga. Se observó que consumir crack es como los otros actos humanos que constituyen el sujeto, aunque se vuelve problemático cuando promueve una "suspensión" de otras posibilidades de vida. Se verificó que tienden a adoptar el discurso normalizado sobre drogas y usuarios, incluso en contradicción con sus historias de vida, aunque que reconocían que hay algo singular en el uso y en sus procesos de tratamiento/recuperación. Esta investigación indica la necesidad de estrategias que promuevan la reflexión de los usuarios sobre el "tipo detrás de la droga", destacando los aspectos universales y singulares de sus biografías, con la construcción de narrativas autobiográficas audiovisuales como herramienta clínicamente poderosa.

A terapêutica de um "CAPS AD" em um coletivo de fotografia / The Therapeutics of a "CAPS AD" in a Photography's Collective

Sendo fundamental a manutenção de um processo de reflexão constante para que a consolidação da Reforma Psiquiátrica possa se dar sem um "engessamento" de práticas que atrapalharia a proposta de atendimento personalizado, adequado à realidade específica de cada caso, faz-se importante observar as experiências que vêm acontecendo nos dispositivos de Atenção Psicossocial. Por isto, este artigo relata experiências ocorridas em um CAPS AD nos anos de 2011 a 2014, narrando o processo de implementação de uma oficina de fotografia e suas implicações. Percebeu-se que, propondo uma atividade aberta, flexível às demandas dos usuários, foi possível despertar interesses artísticos e de gestão, que culminaram no engajamento em um coletivo de fotografia - algo que não se deu por uma obrigatoriedade de frequência a atividades, mas por uma abertura da oficina aos interesses de cada participante, o que funcionou melhor para o aumento da "adesão".(AU)

Considering that it´s fundamental to maintain a process of constant reflection so that the consolidation of the Brazilian Psychiatric Reform can take place without a "plastering" of practices that would hinder the proposal of personalized care, adequate to the specific reality of each case, it is important to observe the experiences that have been happening in the devices of Psychosocial Attention. For this reason, this article reports the experiences that occurred in a CAPS AD in the years 2011 to 2014, narrating the process of implantation of a photography workshop and its implications. It was noticed that, by proposing an open, flexible activity to the user´s demands, it was possible to arouse artistic and management interests, which culminated in the engagement in a photography collective - something that was not due to an obligation to attend activities, but to an opening of the workshop to the interests of each participant, which worked best for increase "membership" and the frequency.(AU)

Characterization of human parainfluenza virus-3 circulating in Israel, 2012-2015.

BACKGROUND: Human parainfluenza virus 3 (hPIV-3) causes respiratory tract infection. OBJECTIVES: The objective of this study was to describe the epidemiology of hPIV-3 infection among hospitalized patients and characterize the circulating strains. STUDY DESIGN: A cross-sectional study was conducted using respiratory samples of 15,946 hospitalized patients with respiratory symptoms in 2012-2015 in Israel. All samples were subjected to q-PCR and q-RT-PCR to determine the presence of hPIV-3 and other respiratory viruses. Samples positive for hPIV-3 were subjected to molecular typing and phylogenetic analysis. RESULTS: Overall, 547 samples 3.4% (95% CI 3.2-3.7) were positive for hPIV-3. Of these 87 (15.9%) were mixed infections; 41.4% with adenovirus, 40.2% with RSV (40.2%) and 19.5% influenza A viruses. The prevalence of hPIV-3 was highest (5.1%) in children aged 0-4 years. Hospitalization in oncology department was associated with increased likelihood of hPIV-3 infection: adjusted odds ratio [aOR] 2.29 (95% confidence intervals [CI] 1.78-2.96), as well as hospitalization in organ transplantation department: aOR 3.65 (95% CI 2.80-4.76). The predominant lineages were C3c (62.3%) and C1b (24.6%), followed by sub-lineages C5 (8.7%) and C3b (2.9%). A new sub-lineage emerged in our analysis, named C1d, which was 17 (1.5%) nucleotide different from C1a, 25 (2.2%) nucleotide different from C1b and 24 (2.1%) nucleotide different from C1c. DISCUSSION: Young children and immunocompromised patients are likely the risk groups for severe respiratory infections with hPIV-3. Strains belonging to lineages C3c and C1b, which are present worldwide, should be targeted in vaccine development. The emergence of new lineage might have public health implications and on vaccine development.

Rapid detection of influenza A pandemic (H1N1) 2009 virus neuraminidase resistance mutation H275Y by real-time reverse transcriptase PCR.

The emergence of oseltamivir-resistant influenza A pandemic (H1N1) 2009 virus highlights the need for rapid oseltamivir resistance screening. We report the development and validation of high-throughput real-time reverse transcriptase PCR assays for the detection of the H275Y substitution in the neuraminidase 1 gene that can be accomplished in 3 to 4 h.

High rate of human bocavirus and adenovirus coinfection in hospitalized Israeli children.

We investigated coinfection of human bocavirus (HBoV) and other respiratory viruses in hospitalized children by real-time PCR. A high rate (69.2%) of adenovirus infection was found among children infected with HBoV. Such high rates of HboV-adenovirus coinfection have not been previously reported, underscoring the need to investigate the contribution of HBoV in patient clinical presentations.

Characterization of human metapneumovirus infections in Israel.

Respiratory tract infections are a leading cause of morbidity and mortality worldwide. Even with the advancement of diagnostic tools, the causative agent of 20 to 30% of upper respiratory tract infections go undiagnosed. Recently, a newly identified human respiratory virus, human metapneumovirus (hMPV), was discovered in young children in The Netherlands. To study the prevalence of hMPV infections in Israeli children, respiratory specimens from 388 hospitalized children less than 5 years of age were evaluated for the presence of hMPV RNA, which was present in 42 (10.8%) of these samples. All hMPV-positive samples were negative for respiratory syncytial virus (RSV), influenza viruses (Flu) A and B, adenovirus, and parainfluenza viruses 1, 2, and 3. Conversely, hMPV RNA was not detected in 76 RSV-positive and 38 Flu A- or B-positive samples. Most hMPV activity was between the months February and April. Sequence analysis of 20 positive samples revealed that both of the hMPV genotypes (groups 1 and 2) have circulated in central Israel during the study period. Moreover, three of the four known hMPV subgroups (1A, 1B, and 2B) were detected among the tested samples. Seroprevalence of hMPV in 204 patients from the central part of Israel revealed that 100% of the children are hMPV seropositive by the age of 5 years old. We conclude that hMPV is a common respiratory pathogen in Israel, while mixed infections of hMPV with RSV or Flu in hospitalized children are apparently rare.

Evaluation of a multiplex real-time reverse transcriptase PCR assay for detection and differentiation of influenza viruses A and B during the 2001-2002 influenza season in Israel.

The ability to rapidly diagnose influenza virus infections is of the utmost importance in the evaluation of patients with upper respiratory tract infections. It is also important for the influenza surveillance activities performed by national influenza centers. In the present study we modified a multiplex real-time reverse transcriptase PCR (RT-PCR) assay (which uses TaqMan chemistry) and evaluated it for its ability to detect and concomitantly differentiate influenza viruses A and B in 370 patient samples collected during the 2001-2002 influenza season in Israel. The performance of the TaqMan assay was compared to those of a multiplex one-step RT-PCR with gel detection, a shell vial immunofluorescence assay, and virus isolation in tissue culture. The TaqMan assay had an excellent sensitivity for the detection of influenza viruses compared to that of tissue culture. The overall sensitivity and specificity of the TaqMan assay compared to the results of culture were 98.4 and 85.5%, respectively. The sensitivity and specificity of the TaqMan assay for the detection of influenza virus A alone were 100 and 91.1%, respectively. On the other hand, the sensitivity and specificity for the detection of influenza virus B alone were 95.7 and 98.7%, respectively. The rapid turnaround time for the performance of the TaqMan assay (4.5 h) and the relatively low direct cost encourage the routine use of this assay in place of tissue culture. We conclude that the multiplex TaqMan assay is highly suitable for the rapid diagnosis of influenza virus infections both in well-established molecular biology laboratories and in reference clinical laboratories.

Specific antibody response after influenza immunization in systemic lupus erythematosus.

OBJECTIVE: To determine the efficacy of influenza virus vaccine in patients with systemic lupus erythematosus (SLE). METHODS: The study population comprised 24 patients with SLE who received the split-virion, inactivated vaccine containing 15 micro g hemagglutinin (HA)/dose of each of A/Beijing/262/95(H1N1), A/Sydney/05/97(H3N2), and B/Harbin/07/94. Hemagglutination inhibition (HI) antibodies were tested using the HI test according to a standard World Health Organization procedure. Immune response was defined as 4-fold or greater rise in HI antibodies 6 weeks after vaccination. Geometric mean titers (GMT) were calculated to assess the immunity of the whole group. RESULTS: All patients were women. Prior to vaccination, the percentage of SLE patients with protective levels of HI antibodies and the GMT of HI antibodies were similar to those of age matched healthy women. Six weeks after vaccination, 75% of the patients had immune response to at least one of the 3 antigens; 58.3% and 62.5% of the patients responded to A/Sydney/05/97(H3N2) and B/Harbin/07/94, respectively. However, only 37.5% of the patients responded to A/Beijing/262/95(H1N1). Six weeks after immunization, the SLE patients generated immune response against a mean number of 1.5 of the 3 influenza vaccines. There was a trend toward a lower immune response in patients with age > or = 50 years, prednisone dosage > or = 10 mg daily, and who used azathioprine. However, methotrexate therapy was not associated with decreased response. CONCLUSION: The immune response to influenza vaccine of patients with SLE is lower than that seen in adults in the general population, in particular among older patients and those treated with immunosuppressive therapy.