ABSTRACT The gold standard for the laboratory diagnosis of COVID-19 is the reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) assay, which searches for SARS-CoV-2 target genes in nasopharyngeal/oropharyngeal (NP/OP) samples, and its performance depends on the quantity and quality of the RNA input. This study compared the performance and cost-effectiveness of three different kits/reagents for RNA extraction used in COVID-19 diagnosis in Sao Paulo, Brazil. A total of 300 NP/OP samples belonging to suspected cases of COVID-19 stored in a biorepository were randomly selected, and RNA was extracted using (i) automated extraction (Loccus, Extracta Kit FAST), (ii) manual extraction (BioGene Kit, Bioclin, Quibasa), and (iii) quick extraction methods (Lucigen, Quick DNA Extract Kit). Next, the samples were tested using RT-qPCR for SARS-CoV-2 with the Allplex 2019-nCoV modified assay and the Charité-Berlin protocol. All assays/kits were used according to the manufacturer’s instructions. For the Allplex kit, the sensitivity in detecting SARS-CoV-2 with previously extracted RNA by different procedures was 100.0% for Loccus, 100.0% for BioGene and 91.9% for Quick. Using the Charité-Berlin protocol, the sensitivities were 81.4% for Loccus, 81.2% for BioGene and 60.7% for Quick. The least sensitive target gene and the gene most affected by RNA extraction procedures was the RNA-dependent RNA polymerase gene (Charité-Berlin protocol). No false-positive SARS-CoV-2 results were detected using RNA obtained from any of the different protocols. In conclusion, Loccus and BioGene RNA extractions were efficient for RT-qPCR assays, and although the BioGene procedure is less expensive, Loccus is the best choice because it allows the rapid handling of hundreds or thousands of samples, a desirable feature during pandemics. Although less sensitive, the Quick extraction is useful during outbreaks coupled with the Allplex amplification kit for SARS-CoV-2 diagnosis (κ = 0.925).

Abstract INTRODUCTION Human retroviruses and the hepatitis B and C viruses (HBV and HCV, respectively) share routes of transmission; thus, coinfections occur and could alter subsequent disease outcomes. A preliminary study on human T-lymphotropic virus types 1 and 2 (HTLV-1/2) in serum samples from HBV- and HCV-infected individuals in São Paulo revealed 1.3% and 5.3% rates of coinfection, respectively. These percentages were of concern since they were detected in HTLV-endemic regions and in high-risk individuals in Brazil. The present study was conducted to extend and confirm these data. METHODS HTLV-1/2 and human immunodeficiency virus (HIV) infection status were identified in 1,984 sera for HBV and HCV viral load quantification - 1,290 samples from HBV-infected individuals (53.3% men, mean age: 47.1 years) and 694 samples from HCV-infected individuals (56.3% men, mean age: 50.1 years). HTLV-1/2 antibodies were detected by enzyme immunoassay, followed by western blotting and line immunoassay; HIV infection was detected by enzyme immunoassay. RESULTS HTLV-1/-2 infection was detected in 1.9% HBV-infected individuals (0.7% HTLV-1 and 1.2% HTLV-2) and in 4.0% (2.4% HTLV-1 and 1.6% HTLV-2) HCV-infected individuals; HIV infection was detected in 9.2% and 14.5%, respectively. Strong associations with HTLV and HIV, male sex, and older age were found in HBV/HTLV and HCV/HTLV-coinfected individuals (p<0.05). CONCLUSIONS HTLV-1 and HTLV-2 were confirmed to be prevalent in individuals with HBV and HCV in São Paulo; coinfected individuals deserve further clinical and laboratory investigation.

RESUMO Objetivo: Comparar o impacto de duas estratégias de fast track quanto ao momento de extubação e retirada da ventilação mecânica invasiva de adultos no pós-operatório cardíaco em desfechos clínicos e hospitalares. Métodos: Estudo de coorte retrospectivo com pacientes submetidos à cirurgia cardíaca. Os pacientes foram classificados de acordo com o tempo de extubação em Grupo Controle (extubados após 6 horas de admissão na unidade de terapia intensiva, com tempo máximo de ventilação mecânica de 18 horas), Grupo 1 (extubados em sala de operação após término da cirurgia) e Grupo 2 (extubados em até 6 horas pós-admissão na unidade de terapia intensiva). Os desfechos primários analisados foram: capacidade vital no primeiro dia de pós-operatório, tempo de internamento hospitalar e na unidade de terapia intensiva. Os desfechos secundários foram reintubação, pneumonia adquirida no hospital, sepse e óbito. Resultados: Para os 223 pacientes avaliados, a capacidade vital foi menor nos Grupos 1 e 2 comparados ao Controle (p = 0,000 e p = 0,046, respectivamente). Os dias de internamento em unidade de terapia intensiva foram significativamente menores nos Grupos 1 e 2 quando comparados ao Controle (p = 0,009 e p = 0,000, respectivamente), já os dias de internamento hospitalar foram menores no Grupo 1 quando comparado ao Controle (p = 0,014). Houve associação entre a extubação na sala de operação (Grupo 1) com reintubação (p = 0,025) e complicações pós-cirúrgicas (p=0,038). Conclusão: Pacientes submetidos ao fast track com extubação em até 6 horas apresentaram menor tempo de internamento em unidade de terapia intensiva sem aumentar complicações pós-cirúrgicas e óbito. Pacientes extubados em sala de operação tiveram menor tempo de internamento hospitalar e em unidade de terapia intensiva, mas apresentaram aumento na frequência de reintubação e complicações pós-cirúrgicas.

ABSTRACT Objective: To compare the impact of two fast-track strategies regarding the extubation time and removal of invasive mechanical ventilation in adults after cardiac surgery on clinical and hospital outcomes. Methods: This was a retrospective cohort study with patients undergoing cardiac surgery. Patients were classified according to the extubation time as the Control Group (extubated 6 hours after admission to the intensive care unit, with a maximum mechanical ventilation time of 18 hours), Group 1 (extubated in the operating room after surgery) and Group 2 (extubated within 6 hours after admission to the intensive care unit). The primary outcomes analyzed were vital capacity on the first postoperative day, length of hospital stay, and length of stay in the intensive care unit. The secondary outcomes were reintubation, hospital-acquired pneumonia, sepsis, and death. Results: For the 223 patients evaluated, the vital capacity was lower in Groups 1 and 2 compared to the Control (p = 0.000 and p = 0.046, respectively). The length of stay in the intensive care unit was significantly lower in Groups 1 and 2 compared to the Control (p = 0.009 and p = 0.000, respectively), whereas the length of hospital stay was lower in Group 1 compared to the Control (p = 0.014). There was an association between extubation in the operating room (Group 1) with reintubation (p = 0.025) and postoperative complications (p = 0.038). Conclusion: Patients undergoing fast-track management with extubation within 6 hours had shorter stays in the intensive care unit without increasing postoperative complications and death. Patients extubated in the operating room had a shorter hospital stay and a shorter stay in the intensive care unit but showed an increase in the frequency of reintubation and postoperative complications.

ABSTRACT Santa Catarina is a Brazilian State that has reported the lowest prevalence of human T-cell lymphotropic viruses (HTLV-1/2) in blood donors (0.04%). Although it presents ports, airports and roads that facilitate the entrance and dissemination of new infectious agents, no information exists concerning the HTLV-1/2 infections in HIV/AIDS patients. This study searched for HTLV-1/2 antibodies in plasma samples of 625 HIV/AIDS patients from the municipality of Tubarao (Southern Santa Catarina), and disclosed 1.1% of positivity (0.48% HTLV-1, 0.48% HTLV-2 and 0.16% untypeable HTLV), and a positive correlation with the male sex (OR 4.16) and intravenous drug use (OR 35.18). Although the percentage of 1.1% appears to be low, it is 27.5 times higher than the percentage detected in blood donors. Since HTLV-1 and HTLV-2 are circulating in HIV-infected individuals in Southern Santa Catarina, and these retroviruses could cause a differently impact on the HIV/AIDS outcomes, the surveillance of HTLV-1/2 is necessary, and it could support public health policies in preventing the transmission and dissemination of these viruses in this State.

ABSTRACT Co-infections of hepatitis C virus (HCV) and either human immunodeficiency virus type 1 (HIV-1), human T-cell lymphotropic virus type 1 (HTLV-1) or type 2 (HTLV-2) have been described as having an impact on HCV viremia and subsequent disease progression. HCV load in serum samples from 622 patients (343 males, 279 females; median age 50.8 years) from São Paulo/southeast Brazil was analyzed using the Abbott Real Time HCV assay (Abbott Molecular Inc., IL, USA). Samples were obtained from HCV-monoinfected (n = 548), HCV/HIV-1- (n = 41), HCV/HTLV-1- (n = 16), HCV/HTLV-2- (n = 8), HCV/HIV/HTLV-1- (n = 4), and HCV/HIV/HTLV-2-co-infected (n = 5) patients, and results were compared among the groups and according to sex. The median HCV load in HCV-monoinfected patients was 5.23 log10 IU/mL and 0.31 log10 higher in men than in women. Increases in viral load of 0.51 log10, 0.54 log10, and 1.43 log10 IU/mL were detected in HCV/HIV-1-, HCV/HTLV-1- and HCV/HIV/HTLV-1-co-infected individuals, respectively, compared with HCV-monoinfected counterparts. In contrast, compared to HCV/HIV co-infected patients, HCV/HTLV-2-co-infected patients had an HCV load of 5.0 log10 IU/mL, whereas HCV/HIV/HTLV-2-co-infected patients had a median load 0.37 log10 IU/mL lower. Significant differences in HCV loads were detected, with males and HCV/HIV-1- and HCV/HIV/HTLV-1-co-infected patients presenting the highest values. Conversely, females and HCV/HTLV-2-co-infected patients exhibited lower HCV loads. Overall, HCV viremia is increased in HIV and/or HTLV-1-co-infection and decreased in HTLV-2 co-infection.

With this study, the authors hope to alert clinicians regarding the presence of human T-cell lymphotropic virus type 1 and 2 (HTLV-1/-2) infections in patients with viral hepatitis B and C in Brazil. HTLV-1/-2 were detected in 1.3% of hepatitis B virus (HBV)- and 5.3% of hepatitis C virus (HCV)-infected blood samples sent for laboratory viral load measurements. A partial association of human immunodeficiency virus (HIV)-1 and HTLV-1/-2 infection was detected in patients with HCV (HIV+, 27.3%), whereas this association was almost 100% in HBV-infected patients (HIV+, all except one). The high prevalence of HTLV-1/-2 infection among patients with hepatitis C was of concern, as HTLV-1/-2 could change the natural course of subsequent liver disease. The authors suggest including HTLV-1/-2 serology in the battery of tests used when following patients with viral hepatitis in Brazil, regardless of the HIV status.

ABSTRACT The present study evaluated several techniques currently available (commercial kits and in-house assays) for diagnosing human T lymphotropic viruses types 1 and 2 in two groups of patients enrolled at HIV/AIDS specialized care services in São Paulo: Group 1 (G1), n = 1608, 1237 male/371 female, median age 44.3 years old, majority using highly active antiretroviral therapy (HAART); G2, n = 1383, 930 male/453 female, median age of 35.6 years old, majority HAART naïve. Enzyme immunoassays [(EIA) Murex and Gold ELISA] were employed for human T lymphotropic viruses types 1 and 2 screening; Western blotting (WB), INNO-LIA (LIA), real-time PCR pol (qPCR), and nested-PCR-RFLP (tax) were used to confirm infection. Samples were considered human T lymphotropic viruses types 1 and 2 positive when there was reactivity using at least one of the four confirmatory assays. By serological screening, 127/2991 samples were positive or borderline, and human T lymphotropic virus infection was confirmed in 108 samples (three EIA-borderline): 56 human T lymphotropic virus type 1 [G1 (27) + G2 (29)]; 45 human T lymphotropic virus type 2 [G1 (21) + G2 (24)]; one human T lymphotropic virus type 1 + human T lymphotropic virus type 2 (G2); six human T lymphotropic virus [G1 (2) + G2 (4)]. Although there were differences in group characteristics, human T lymphotropic viruses types 1 and 2 prevalence was similar [3.1% (G1) and 4.2% (G2), p = 0.113]. The overall sensitivities of LIA, WB, qPCR, and PCR-RFLP were 97.2%, 82.4%, 68.9%, and 68.4%, respectively, with some differences among groups, likely due to the stage of human T lymphotropic virus infection and/or HAART duration. Indeterminate immunoblotting results were detected in G2, possibly due to the seroconversion period. Negative results in molecular assays could be explained by the use of HAART, the occurrence of defective provirus and/or the low circulating proviral load. In conclusion, when determining the human T lymphotropic virus infection, the findings highlight that there is a need to consider the blood samples with borderline results in screening assays. Of all the tested assays, LIA was the assay of choice for detecting human T lymphotropic virus type 1 and human T lymphotropic virus type 2 in human immunodeficiency virus type 1-infected patients.

OBJETIVO: apresentar o percurso seguido pela Fonoaudiologia no estado de Minas Gerais desde a chegada dos primeiros profissionais até os dias de hoje. MÉTODO: trata-se de pesquisa de campo, exploratória e histórica, de abordagem qualiquantitativa. Participaram 27 profissionais, fonoaudiólogos e médicos envolvidos no processo de implantação, organização e gestão de serviços, cursos e consultórios de Fonoaudiologia. A coleta de dados ocorreu por meio do levantamento de documentos e entrevistas semi-estruturadas. Após essa fase, houve a categorização dos documentos e das entrevistas. RESULTADOS: o relato do histórico se inicia com a chegada dos primeiros fonoaudiólogos em Minas Gerais. A descrição das entrevistas se divide nas seguintes categorias: contato com a Fonoaudiologia; motivação para a escolha da profissão; inserção na Fonoaudiologia; motivação para o trabalho em Minas; contexto da Fonoaudiologia em Minas Gerais nas décadas de 70 a 90; formação profissional; impulsos para o reconhecimento e avanços da profissão; perspectivas da Fonoaudiologia; avaliação do estudo histórico. CONCLUSÃO: foi possível demonstrar o caminho que a Fonoaudiologia traçou no estado de Minas Gerais desde a chegada dos primeiros fonoaudiólogos até os dias de hoje.

PURPOSE: to present the course traced by Speech-language Pathology in Minas Gerais state since the arrival of the first professionals up to the current time. METHOD: it is an exploratory and historical field research based on a quality-quantitative approach. Twenty-seven professionals including speech-language pathologists and doctors involved in the process of implementing, organizing and managing services, courses and Speech-language pathology offices took part in this study. Data were collected by using documents research and semi-structured interviews. Then, there was a categorization of the documents and interviews. RESULTS: the historical report begins with the arrival of the first speech-language pathologists in Minas Gerais. The description of the interviews is divided in the following categories: first time in touch with Speech-language pathology; motivation for choosing this professional occupation; insertion in Speech-language pathology; motivation for working in Minas Gerais; context of this science from the 1970s to the 1990s; career development; boost to the recognition and progress of the profession; Speech-language pathology perspectives; evaluation of the historical study. CONCLUSION: it was possible to demonstrate the course traced by Speech-language Pathology in Minas Gerais province since the arrival of the first professionals up to the current time.