Rapid response of a public health reference laboratory to the COVID-19 pandemic.

Introduction. Brazil was one of the most affected countries by the COVID-19 pandemic. Instituto Adolfo Lutz (IAL) is the reference laboratory for COVID-19 in São Paulo, the most populous state in Brazil. In April 2020, a secondary diagnostic pole named IAL-2 was created to enhance IAL's capacity for COVID-19 diagnosis.Hypothesis/Gap Statement. Public health laboratories must be prepared to rapidly respond to emerging epidemics or pandemics.Aim. To describe the design of IAL-2 and correlate the results of RT-qPCR tests for COVID-19 with secondary data on suspected cases of SARS-CoV-2 infection in the São Paulo state.Methodology. This is a retrospective study based on the analysis of secondary data from patients suspected of infection by SARS-CoV-2 whose clinical samples were submitted to real-time PCR after reverse transcription (RT-qPCR) at IAL-2, between 1 April 2020 and 8 March 2022. RT-qPCR Ct results of the different kits used were also analysed.Results. IAL-2 was implemented in April 2020, just over a month after the detection of the first COVID-19 case in Brazil. The laboratory performed 304,250 RT-qPCR tests during the study period, of which 98 319 (32.3 %) were positive, 205827 (67.7 %) negative, and 104 (0.03 %) inconclusive for SARS-CoV-2. RT-qPCR Ct values≤30 for E/N genes of SARS-CoV-2 were presented by 79.7 % of all the samples included in the study.Conclusion. IAL was able to rapidly implement a new laboratory structure to support the processing of an enormous number of samples for diagnosis of COVID-19, outlining strategies to carry out work with quality, using different RT-qPCR protocols.

Detection of herpesviruses in neotropical primates from São Paulo, Brazil.

Transmission of herpesvirus between humans and non-human primates represents a serious potential threat to human health and endangered species conservation. This study aimed to identify herpesvirus genomes in samples of neotropical primates (NTPs) in the state of São Paulo, Brazil. A total of 242 NTPs, including Callithrix sp., Alouatta sp., Sapajus sp., and Callicebus sp., were evaluated by pan-herpesvirus polymerase chain reaction (PCR) and sequencing. Sixty-two (25.6%) samples containing genome segments representative of members of the family Herpesviridae, including 16.1% for Callitrichine gammaherpesvirus 3, 6.1% for Human alphaherpesvirus 1, 2.1% for Alouatta macconnelli cytomegalovirus, and 0.83% for Cebus albifrons lymphocryptovirus 1. No co-infections were detected. The detection of herpesvirus genomes was significantly higher among adult animals (p = 0.033) and those kept under human care (p = 0.008671). These findings confirm the importance of monitoring the occurrence of herpesviruses in NTP populations in epizootic events.

Concurrent yellow fever and pulmonary aspergillosis due to Aspergillus fumigatus in a free-ranging howler monkey (Alouatta sp).

Herein, we describe a unique case of concomitant angioinvasive pulmonary aspergillosis due to Aspergillus fumigatus and yellow fever in a free-ranging howler monkey (Alouatta sp). Lung samples were negative for influenza viruses A and B.

Human Cytomegalovirus DNA Quantification and Gene Expression in Gliomas of Different Grades.

Gliomas are the most common type of primary brain tumors. The most aggressive type, Glioblastoma multiforme (GBM), is one of the deadliest human diseases, with an average survival at diagnosis of about 1 year. Previous evidence suggests a link between human cytomegalovirus (HCMV) and gliomas. HCMV has been shown to be present in these tumors and several viral proteins can have oncogenic properties in glioma cells. Here we have investigated the presence of HCMV DNA, RNA and proteins in fifty-two gliomas of different grades of malignancy. The UL83 viral region, the early beta 2.7 RNA and viral protein were detected in 73%, 36% and 57% by qPCR, ISH and IHC, respectively. Positivity of the viral targets and viral load was independent of tumor type or grade suggesting no correlation between viral presence and tumor progression. Our results demonstrate high prevalence of the virus in gliomas from Brazilian patients, contributing to a better understanding of the association between HCMV infection and gliomas worldwide and supporting further investigations of the virus oncomodulatory properties.

Avaliação do exame imuno-histoquímico para o diagnóstico de Leishmania spp. em amostras de tecidos caninos / Evaluation of immunohistochemical technique for the diagnosis of Leishmania spp. in canine tissues samples

No Brasil, as leishmanioses representam um importante problema de saúde pública pela sua elevada incidência, ampla distribuição geográfica e marcante aumento na transmissão associados à urbanização da doença. Cães são considerados os principais reservatórios da leishmaniose visceral (LV) no ambiente urbano, tornando-se fundamental o aprimoramento do diagnóstico dadoença nessa espécie. Este estudo objetivou a avaliação da técnica de imuno-histoquímica (IHQ) para o diagnóstico de Leishmania spp. em amostras de diferentes tecidos de cães recebidas no Núcleo de Patologia Quantitativa do Instituto Adolfo Lutz (NPQ-IAL). Amostras de tecidos coletadas de 134 cães, provenientes de municípios do Estado de São Paulo, positivos para LV por teste rápido(TR DPP®-Bio-Manguinhos), ensaio imunoenzimático-EIE/Elisa (Bio-Manguinhos) e pela reação em cadeia de polimerase, foram submetidas a marcação IHQ específica, em duplicata. A reação de IHQ apresentou sensibilidade de 98,51 %, especificidade de 100,00 % e acurácia de 98,61 %. A maior positividade foi detectada nas amostras de baço. O índice Kappa foi de 0,84 entre os resultados da análise dos tecidos em duplicata. A técnica de IHQ pode ser utilizada como uma técnica rotineirapara o diagnóstico das leishmanioses caninas, sendo padronizada e implantada no NPQ-IAL.

In Brazil, leishmaniasis represents an important public health problem due to its high incidence, wide geographic distribution and marked increase in transmission associated to the urbanization of the disease. Dogs are considered the main reservoirs of visceral leishmaniasis (VL) in the urban environment, making it fundamental to improve the diagnosis of this species. This study aimed at the evaluation of the immunohistochemical technique (IHC) for the diagnosis of Leishmania spp. In samples of different dog tissues received at the Adolfo Lutz Institute's Quantitative Pathology Center (NPQ-IAL). Tissue specimens collected from 134 dogs, from municipalities in the State of São Paulo, positive for LV by rapid test (TR DPP®-Bio-Manguinhos), enzyme-linked immunosorbent assay (EIE / Elisa (Bio-Manguinhos) and the chain reaction of Polymerase, were subjected to specific IHQ labeling, in duplicate. The IHQ reaction showed sensitivity of 98.51%, specificity of 100.00% and accuracy of 98.61%. The highest positivity was detected in spleen samples. The Kappa index was 0.84 between the results of the tissue analysis in duplicate. The IHC technique can be used as a routine technique for the diagnosis of canine leishmaniasis, being standardized and implanted in the NPQ-IAL.

Human Cytomegalovirus DNA Quantification and Gene Expression in Gliomas of Different Grades

Gliomas are the most common type of primary brain tumors. The most aggressive type, Glioblastoma multiforme (GBM), is one of the deadliest human diseases, with an average survival at diagnosis of about 1 year. Previous evidence suggests a link between human cytomegalovirus (HCMV) and gliomas. HCMV has been shown to be present in these tumors and several viral proteins can have oncogenic properties in glioma cells. Here we have investigated the presence of HCMV DNA, RNA and proteins in fifty-two gliomas of different grades of malignancy. The UL83 viral region, the early beta 2.7 RNA and viral protein were detected in 73%, 36% and 57% by qPCR, ISH and IHC, respectively. Positivity of the viral targets and viral load was independent of tumor type or grade suggesting no correlation between viral presence and tumor progression. Our results demonstrate high prevalence of the virus in gliomas from Brazilian patients, contributing to a better understanding of the association between HCMV infection and gliomas worldwide and supporting further investigations of the virus oncomodulatory properties.

Evaluation of immunohistochemical technique for the diagnosis of Leishmania spp. in canine tissues samples / Avaliação do exame imuno-histoquímico para o diagnóstico de Leishmania spp. em amostras de tecidos caninos

In Brazil, leishmaniasis represents a major public health problem due to its high incidence, wide distribution and remarkable increase in transmission associated with the disease urbanization. Dogs are considered as the main reservoirs of visceral leishmaniasis (VL) in the urban environment and diagnosis improvement was fundamental in this species. This study aimed at evaluating the immunohistochemical technique (IHC) for diagnosing Leishmania spp. in differents tissues samples collected from dogs. This investigation was performed at the Quantitative Pathology Center of the Institute Adolfo Lutz (NPQ-IAL). Tissue samples collected from 134 dogs, derived from municipalities of São Paulo State, positive for VL by rapid testing (TR DPP® - Bio-Manguinhos), enzyme-linked immunosorbent assay (EIA/ELISA - Bio-Manguinhos), and polymerase chain reaction, were analyzed by specific IHC in duplicate. These dogs were previously diagnosed as VL. IHC showed sensitivity of 98.51 %, specificity of 100.00 %, positive predictive value of 100.00 %, negative predictive value of 83.33 % and accuracy of 98.61 %. The major positivity was detected in spleen samples. Kappa index between the tissues duplicate results was of 0.84. IHC technique, standardized and implemented at NPQ-IAL, showed to be suitable to be used as routine test for diagnosing canine leishmaniasis effectivelly.

No Brasil, as leishmanioses representam um importante problema de saúde pública pela sua elevada incidência, ampla distribuição geográfica e marcante aumento na transmissão associados à urbanização da doença. Cães são considerados os principais reservatórios da leishmaniose visceral (LV) no ambiente urbano, tornando-se fundamental o aprimoramento do diagnóstico da doença nessa espécie. Este estudo objetivou a avaliação da técnica de imuno-histoquímica (IHQ) para o diagnóstico de Leishmania spp. em amostras de diferentes tecidos de cães recebidas no Núcleo de Patologia Quantitativa do Instituto Adolfo Lutz (NPQ-IAL). Amostras de tecidos coletadas de 134 cães, provenientes de municípios do Estado de São Paulo, positivos para LV por teste rápido (TR DPP®-Bio-Manguinhos), ensaio imunoenzimático-EIE/Elisa (Bio-Manguinhos) e pela reação em cadeia de polimerase, foram submetidas a marcação IHQ específica, em duplicata. A reação de IHQ apresentou sensibilidade de 98,51 %, especificidade de 100,00 % e acurácia de 98,61 %. A maior positividade foi detectada nas amostras de baço. O índice Kappa foi de 0,84 entre os resultados da análise dos tecidos em duplicata. A técnica de IHQ pode ser utilizada como uma técnica rotineira para o diagnóstico das leishmanioses caninas, sendo padronizada e implantada no NPQ-IAL.

Biorrepositórios e Biobancos: inovação estratégica em Saúde Pública / Biorepository and Biobank: strategic inovation in the Public Health Service

Os avanços tecnológicos das últimas décadas catalisaram transformações sociais e econômicas, que influenciaram decisivamente nos padrões da morbimortalidade populacional. No Brasil, a heterogeneidade deste padrão é muito visível e complexa, em função de sua grande extensão territorial, do significativo número de habitantes e das diferenças socioeconômicas e culturais. Com o aumento da expectativa devida e o envelhecimento da população brasileira, observa-se o aparecimento cada vez mais frequente de doenças crônicas não transmissíveis. As mudanças climáticas e as condições higiênico-sanitárias, ainda deficientes em algumas regiões, podem propiciar o recrudescimento das doenças infectocontagiosas. Em face dessas transformações, faz-se necessário que o sistema de vigilância epidemiológica seja reestruturado para adequar aos novos cenários epidemiológicos, identificar novos riscos, prever e conter a expansão de áreas com riscos preexistentes de disseminação, propagação e redução das doenças. Um dos pontos cruciais desta reestruturação é o armazenamento correto e adequado das amostras biológicas, por meio da criação de biorrepositórios e biobancos, que possibilitará a aplicação de novas tecnologias para detecção, investigação e respostas às situações de surtos, epidemias e pandemias, com benefícios à pesquisa e assistência na área de saúde. Esta revisão demonstra a importância dos biobancos e biorrepositórios em Saúde Pública.

A pesquisa do vírus de Epstein-Barr (EBV) pela reação de hibridização in situ realizada no Núcleo de Patologia Quantitativa - Centro de Patologia - IAL / Study of the Epstein-Barr virus (EBV) by in situ hybridization performed at Laboratory of Quantitative Pathology - Center of Pathology - Adolfo Lutz Intitute

O objetivo deste estudo foi analisar os dados preliminares da detecção do Epstein-Barr Virus (EBV) pela reação de hibridização in situ cromogênica (CISH) nas amostras encaminhadas ao Núcleo de Patologia Quantitativa/Instituto Adolfo Lutz. Os 12 casos com suspeita da presença do EBV foram recebidos no período de outubro de 2011 a julhode 2012, totalizando 14 amostras, sendo que quatro casos apresentavam resultados prévios pelo estudo imuno-histoquímico (IHQ): três negativos e um positivo para o EBV LMP-1. Foi utilizado o Zytofast EBV Probe e o ZytoFast Plus CISH Implementation Kit – HRP – DAB adaptado às condições do laboratório. Apositividade para EBER (Epstein-Barr Encoded Early RNAs) do EBV foi observada em 10 amostras, inclusive nos três casos previamente negativos pelo exame IHQ. Concluímos, neste estudo preliminar, que a reação de CISH permitiu detectar sequências gênicas do EBV, mesmo em amostras negativas para proteína latente de membrana (LMP-1) pela reação IHQ