In-Depth Longitudinal Comparison of Clinical Specimens to Detect SARS-CoV-2

The testing and isolation of patients with coronavirus disease 2019 (COVID-19) are indispensable tools to control the ongoing COVID-19 pandemic. PCR tests are considered the “gold standard” of COVID-19 testing and mostly involve testing nasopharyngeal swab specimens. Our study aimed to compare the sensitivity of tests for various sample specimens. Seventy-five participants with confirmed COVID-19 were included in the study. Nasopharyngeal swabs, oropharyngeal swabs, Oracol-collected saliva, throat washes and rectal specimens were collected along with pooled swabs. Participants were asked to complete a questionnaire to correlate specific clinical symptoms and the symptom duration with the sensitivity of detecting COVID-19 in various sample specimens. Sampling was repeated after 7 to 10 days (T2), then after 14 to 20 days (T3) to perform a longitudinal analysis of sample specimen sensitivity. At the first time point, the highest percentages of SARS-CoV-2-positive samples were observed for nasopharyngeal samples (84.3%), while 74%, 68.2%, 58.8% and 3.5% of throat washing, Oracol-collected saliva, oropharyngeal and rectal samples tested positive, respectively. The sensitivity of all sampling methods except throat wash samples decreased rapidly at later time points compared to the first collection. The throat washing method exhibited better performance than the gold standard nasopharyngeal swab at the second and third time points after the first positive test date. Nasopharyngeal swabs were the most sensitive specimens for early detection after symptom onset. Throat washing is a sensitive alternative method. It was found that SARS-CoV-2 persists longer in the throat and saliva than in the nasopharynx.

SARS-CoV-2 RNA detection using pooling of self-collected samples: Simple protocol may foster asymptomatic surveillance

1AbstractBackgroundSurveillance of COVID infection and isolation of infected individuals is one of the available tools to control the spread of SAR-CoV-2. Asymptomatic and pre symptomatic are responsible for substantial transmission. RNA or antigen tests are necessary to identify non-symptomatic individuals. We tested the feasibility of using samples pooling offering different collection alternatives (swab/throat wash/saliva) to volunteers of a public health institute.MethodsWe evaluated pool samples from frozen material from previously tested samples and a prospective collection from asymptomatic volunteers. Some collections were paired for comparison. Pools and some individual samples were extracted with QIAamp Viral RNA Mini Kit (Qiagen, USA) and/or Lucigen Quick Extract DNA extraction solution (BioSearch, USA) and submitted to rtPCR (Allplex, Seegene, Korea).ResultsA total of 240 samples from 130 new collections and 37 samples with known result were evaluated. Pool CT was generally higher than individual samples. Lucigen extraction showed higher CT, including false negative results for samples with high CT at Qiagen extraction. Paired Swab and TW samples showed comparable results. No volunteer from negative pools reported any symptom in the 2-3 days after collection.ConclusionsClinical samples pooling to detect SARS-CoV-2 RNA is feasible and an economical way to test for COVID-19, especially in surveillance strategies targeting more infectiousness, higher viremia individuals. The use of Lucigen reagents show lower sensibility that may lead to false negative results with lower viremia samples. Combining throat wash with saliva may provide and interesting self-collection alternative, but more comparative work is needed.

Recomendações em cirurgias bucomaxilofaciais de urgência e emergência em tempos de COVID-19

Embora as rotas de transmissão da COVID-19 ainda não estejam totalmente identificadas, a transmissão homem-a-homem já é uma realidade em muitos países. Este trabalho tem por objetivo reunir recomendações dos órgãos de saúde e de estudos em artigos científicos relacionadas a condutas especiais do cirurgião bucomaxilofacial no período perioperatório de urgência e emergência. A justificativa é a possibilidade de infecção cruzada por meio da inalação de partículas produzidas durante os procedimentos cirúrgicos. A abordagem desse trabalho está relacionada à rota de transmissão e a possibilidade de contaminação cruzada da COVID-19 por meio de gotículas de salivas transformadas em aerossóis. Considera-se que a adoção de condutas especiais no período periorperatório por parte do cirurgião bucomaxilofacial pode representar auxílio quanto a prevenção de transmissão do vírus tanto em relação aos profissionais quanto aos pacientes.Descritores: Infecções por Coronavírus; Assistência Perioperatória; Aerossóis; Transmissão de Doença Infecciosa do Paciente para o Profissional.ReferênciasShoji H, Fonseca EKUN, Teles GBDS, et al. Structured thoracic computed tomography report for COVID-19 pandemic. Einstein (Sao Paulo). 2020;18:eED5720.Wu Z, McGoogan JM. Characteristics of and Important Lessons From the Coronavirus Disease 2019 (COVID-19) Outbreak in China: Summary of a Report of 72 314 Cases From the Chinese Center for Disease Control and Prevention. JAMA. 2020;323(13):1239-42.Sabino-Silva R, Jardim ACG, Siqueira WL. Coronavirus COVID-19 impacts to dentistry and potential salivary diagnosis. Clin Oral Investig. 2020;24(4):1619-21.Rodrigues MFB, Rocha LLDA, Acioly RDF, Souza DDD, Carvalho DDC, Rocha RCLD, Rocha CCLD. Special precautions in oral and maxillofacial surgeries regarding COVID-19 transmission. Preprints 2020, 2020050135 (doi: 10.20944/preprints202005.0135.v1).To KKW, Tsang OTY, Leung WS, Tam AR, Wu TC, Lung DC et al. Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study. Lancet Infect Dis. 2020;20(5):565-74.Ather A, Patel B, Ruparel NB, Diogenes A, Hargreaves KM. Coronavirus Disease 19 (COVID-19): Implications for Clinical Dental Care. J Endod. 2020;46(5):584-95.Meng L, Hua F, Bian Z. Coronavirus Disease 2019 (COVID-19): Emerging and Future Challenges for Dental and Oral Medicine. J Dent Res. 2020;99(5):481-87.Ge ZY, Yang LM, Xia JJ, Fu XH, Zhang YZ. Possible aerosol transmission of COVID-19 and special precautions in dentistry. J Zhejiang Univ Sci B. 2020;21(5):361-68. Wang WK, Chen SY, Liu IJ, Chen YC, Chen HL, Yang CF et al. Detection of SARS-associated coronavirus in throat wash and saliva in early diagnosis. Emerg Infect Dis. 2004;10(7):1213-19.Chang L, Yan Y, Wang L. Coronavirus Disease 2019: Coronaviruses and Blood Safety. Transfus Med Rev. 2020;34(2):75-80.Spagnuolo G, De Vito D, Rengo S, Tatullo M. COVID-19 Outbreak: An Overview on Dentistry. Int J Environ Res Public Health. 2020;17(6):2094.Zucco L, Levy N, Ketchandji D, Aziz M, Ramachandran SK. Considerações perioperatórias para o novo coronavírus 2019 (COVID-19). Rochester: ASPF; 2020.Abosadegh MM, Saddki N, Al-Tayar B, Rahman SA. Epidemiology of maxillofacial fractures at a tea- ching hospital in Malaysia: a retrospective study. BioMed Res Int. 2019: 9024763.Colégio Brasileiro de Cirurgia e Traumatologia Bucomaxilofacial (CTBMF). COVID-19 - Guia de Práticas em CTBMF. Disponível em: https://www.bucomaxilo.org.br/site/noticiasdetalhes.php?cod=344amp;q=COVID19%2B%20%2BGuia%2Bde%2BPr%C3%A1ticas%2Bem%2BCTBMFamp;bsc=ativar. Acesso em: 07 de abr. de 2020.American Association of Oral and Maxillofacial Surgeons (AAOMS). Member Alert: COVID-19 Guidance for OMS Practices. Disponível em: https://www.aaoms.org/news/member-alert-covid-19-guidance-for-oms-practices. Acesso em: 10 de fev. de 2020.AOCMF International Task Force Recommendations on Best Practices for Maxillofacial Procedures during COVID-19 Pandemic. Disponível em: https://aocmf3.aofoundation.org/-/media/project/aocmf/aocmf/files/covid-19/ao_cmf_covid-19_task_force_guidelines.pdf. Acesso em: 10 de fev. de 2020.

Cuidados especiais na área de cirurgia e traumatologia bucomaxilofacial frente à COVID-19

A organização mundial de saúde definiu o surto do novo coronavírus como uma emergência de saúde pública de interesse internacional. A média de idade da ocorrência da doença ocasionada pelo vírus está na faixa de 49 a 59 anos. Os sintomas da infecção da COVID-19 incluem febre, tosse e doença respiratória aguda. Muitos procedimentos bucomaxilofaciais hospitalares produzem aerossóis e gotículas contaminados por sangue, bactérias e vírus. O objetivo desse estudo é reunir recomendações de órgãos de saúde e artigos científicos para orientação do cirurgião quanto aos procedimentos de atendimento e tratamentos em cirurgia bucomaxilofacial. A finalidade é prevenir a transmissão da Covid-19 durante o tratamento de paciente em situação de urgência e emergência. A metodologia utilizou as orientações do Colégio Brasileiro de Cirurgia e Traumatologia Buco-maxilo-facial, além de recomendações e dados epidemiológicos de órgãos de saúde nacionais e internacionais e artigos científicos publicados. Os cirurgiões bucomaxilofaciais, por natureza correm alto risco de exposição a doenças infecciosas. O surgimento da COVID-19 impôs novos desafios quanto a compreensão da transmissão do vírus por meio de gotículas de saliva e aerossóis.Descritores: Infecções por Coronavirus; Emergências; Procedimentos Cirúrgicos Bucais.ReferênciasGuo H, Zhou Y, Liu X, Tan J. The impact of the COVID-19 epidemic on the utilization of emergency dental services. J Dent Sci. 2020;10.1016/j.jds.2020.02.002.Ge ZY, Yang LM, Xia JJ, Fu XH, Zhang YZ. Possible aerosol transmission of COVID-19 and special precautions in dentistry. J Zhejiang Univ Sci B. 2020;21(5):361-68. Rodrigues MFB, Rocha LLDA, Acioly RDF, Souza DDD, Carvalho DDC, Rocha RCLD, Rocha CCLD. Special precautions in oral and maxillofacial surgeries regarding COVID-19 transmission. Preprints 2020, 2020050135 (doi: 10.20944/preprints202005.0135.v1).Meng L, Hua F, Bian Z. Coronavirus Disease 2019 (COVID-19): Emerging and Future Challenges for Dental and Oral Medicine. J Dent Res. 2020;99(5):481-87.Sabino-Silva R, Jardim ACG, Siqueira WL. Coronavirus COVID-19 impacts to dentistry and potential salivary diagnosis. Clin Oral Investig. 2020;24(4):1619-21. Tuñas ITC, Silva ET, Santiago SBS, Maia KD, Silva-Júnior GO. Doença pelo Coronavírus 2019 (COVID-19): Uma abordagem preventiva para Odontologia. Rev Bras Odontol. 2020;77(1):1-6.Spagnuolo G, De Vito D, Rengo S, Tatullo M. COVID-19 Outbreak: An Overview on Dentistry. Int J Environ Res Public Health. 2020;17(6):2094.Yang Y, Lu Q, Liu M, Wang Y, Zhang A, Jalali N et al. Epidemiological and clinical features of the 2019 novel coronavirus outbreak in China. medRxiv 2020. doi: https://doi.org/ 10.1101/2020.02.10.20021675.To KKW, Tsang OTY, Leung WS, Tam AR, Wu TC, Lung DC et al. Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study. Lancet Infect Dis. 2020;20(5):565-74.Wang WK, Chen SY, Liu IJ, Chen YC, Chen HL, Yang CF et al. Detection of SARS-associated coronavirus in throat wash and saliva in early diagnosis. Emerg Infect Dis. 2004;10(7):1213-19.Associação de Medicina Intensiva Brasileira. Conselho Federal de Odontologgia. Recomendações A, para atendimento odontológico COVID C. Comitê de Odontologia AMIB/CFO de enfrentamento ao COVID-19 Departamento de Odontologia AMIB–1 Atualização 25/03/2020. São Paulo: AMIB; 2020.Brasil. Agência Nacional de Vigilância Sanitária. Nota técnica GVIMS/GGTES/ANVISA Nº 04/2020. Orientações para Serviços de Saúde: Medidas de Prevenção e Controle que devem ser adotadas durante a assistência aos casos suspeitos ou confirmados de infecção pelo novo Coronavírus (SARS-COV-2). Brasília: ANVISA; 2020.Colégio Brasileiro de Cirurgia e Traumatologia Bucomaxilofacial. COVID-19 - Guia de Práticas em CTBMF. Disponível em: https://www.bucomaxilo.org.br/site/noticiasdetalhes.php?cod=344amp;q=COVID19%2B%20%2BGuia%2Bde%2BPr%C3%A1ticas%2Bem%2BCTBMFamp;bsc=ativar. Acesso em: 07 de abr. de 2020.

Throat wash as a source of SARS-CoV-2 RNA to monitor community spread of COVID-19.

Background: SARS-CoV-2 RNA detection with real time PCR is currently the central diagnostic tool to determine ongoing active infection. Nasopharyngeal and oral swabs are the main collection tool of biological material used as the source of viral RNA outside a hospital setting. However, limitation in swabs availability, trained health professional with proper PPE and potential risk of aerosols may hinder COVID diagnosis. Self-collection with swabs, saliva and throat wash to obtain oropharyngeal wash has been suggested as having comparable performance of regular swab. We performed throat wash (TW) based surveillance with laboratory heath workers and other employees (LHW) at a laboratory research institute. Methods: Consecutive volunteer testing of LWH and external household and close contacts were included. TW self-collection was performed in 5 mL of sterile saline that was returned to original vial after approximate 5 secs of gargle. RNA extraction and rtPCR were performed as part of routine COVID protocols using Allplex (Seegene, Korea). Results: Four hundred and twenty two volunteers, 387 (93%) LHW and 43 (7%) contacts participated in the survey. One or more positive COVID rtPCR was documented in 63 (14.9% CI95 12%-19%) individuals. No correlation was observed between with direct activities with COVID samples to positivity, with infection observed in comparable rates among different laboratory areas, administrative or supportive activities. Among 63 with detected SARS-CoV-2 RNA, 59 with clinical information, 58% reported symptoms at a median of 4 days prior to collection, most with mild disease. Over a third (38%) of asymptomatic cases developed symptoms 1-3 days after collection. Although overall CT values of TW were higher than that of contemporary swab tests from hospitalized cases, TW from symptomatic cases had comparable CTs. Conclusions: The study suggests that TW may be a valid alternative to the detection of SARS-CoV-2 RNA. The proportion of asymptomatic and pre-symptomatic cases is elevated and reinforces the need of universal precautions and frequent surveys to limit the spread of the disease.