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au:SANTI, SILVIA MARIA DI

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An algorithm based on molecular protocols to improve the detection of Plasmodium in autochthonous malarial areas in the Atlantic Forest biome
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Farinas, Maria de Lourdes Rego Neves

; Aschar, Mariana

; Costa-Nascimento, Maria de Jesus

; Di Santi, Silvia Maria

Revista do Instituto de Medicina Tropical de São Paulo

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2022, Volume 64 elocation e18

Abstract: En | Text: En | PDF: En

ABSTRACT Malaria is the most important vector-borne disease in the world and a challenge for control programs. In Brazil, 99% of cases occur in the Amazon region. In the extra-Amazonian region, a non-endemic area, epidemiological surveillance focuses on imported malaria and on autochthonous outbreaks, including cases with mild symptoms and low parasitemia acquired in the Atlantic Forest biome. In this scenario, cases are likely to be underreported, since submicroscopic parasitemias are not detected by thick blood smear, considered the reference test. Molecular tests are more sensitive, detecting asymptomatic individuals and mixed infections. The aim of this study was to propose a more efficient alternative to detect asymptomatic individuals living in areas of low malaria endemicity, as they are reservoirs of Plasmodium that maintain transmission locally. In total, 955 blood samples from residents of 16 municipalities with autochthonous malaria outbreaks in the Sao Paulo State were analyzed; 371 samples were collected in EDTA tubes and 584 in filter paper. All samples were initially screened by a genus-specific qPCR targeting ssrRNA genes (limit of detection of 1 parasite/µL). Then, positive samples were subjected to a nested PCR targeting ssrRNA and dihydrofolate reductase-thymidylate synthase genes (limit of detection of 10 parasites/µL) to determine Plasmodium species. The results showed a statistically significant difference (K = 0.049; p < 0.0001) between microscopy positivity (6.9%) and qPCR (22.9%) for EDTA-blood samples. Conversely, for samples collected in filter paper, no statistical difference was observed, with 2.6% positivity by thick blood smear and 3.1% for qPCR (K = 0.036; p = 0.7). Samples positive by qPCR were assayed by a species-specific nested PCR that was in turn positive in 26% of samples (16 P. vivax and 4 P. malariae ). The results showed that molecular protocols applied to blood samples from residents in areas with autochthonous transmission of malaria were useful to detect asymptomatic patients who act as a source of transmission. The results showed that the genus-specific qPCR was useful for screening positives, with the subsequent identification of species by nested PCR. Additional improvements, such as standardization of blood plotting on filter paper and a more sensitive protocol for species determination, are essential. The qPCR-based algorithm for screening positives followed by nested PCR will contribute to more efficient control of malaria transmission, offering faster and more sensitive tools to detect asymptomatic Plasmodium reservoirs.

https://doi.org/10.1590/s1678-9946202264018

Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories
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Aschar, Mariana

; Sanchez, Maria Carmen A.

; Costa-Nascimento, Maria de Jesus

; Farinas, Maria de Lourdes R. N.

; Hristov, Angélica D.

; Lima, Giselle F. M. C.

; Inoue, Juliana

; Levi, José E.

; Di Santi, Silvia M.

Revista Panamericana de Salud Pública

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2022, Volume 46 elocation e11

Abstract: Pt En Es | Text: Pt En Es | PDF: Pt | PDF: En | PDF: Es

RESUMO Objectivo. Avaliar ferramentas moleculares para detectar parasitemia de baixo nível e as cinco espécies de Plasmodium que infectam humanos, para utilização em programas de controlo e eliminação e em laboratórios de referência. Métodos. Avaliámos 145 amostras de sangue de doentes que testaram positivo por reacção em cadeia da polimerase aninhada (nPCR), de indivíduos assintomáticos, e do Programa Global de Paludismo da Organização Mundial de Saúde/Serviço Nacional de Avaliação da Qualidade Externa do Reino Unido. As amostras foram ensaiadas utilizando o RealStar® Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) e o RealStar® Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). Os resultados dos testes moleculares foram comparados com os resultados da PCR quantitativa (qPCR), nPCR e exame da gota espessa. Resultados. Os níveis de parasitemia variaram de 1 a 518 000 parasitas/µL, dependendo da espécie. Em comparação com a nPCR, alt-S&T tinha uma sensibilidade de 100%, excepto na identificação de P. falciparum, para a qual a sensibilidade era de 93,94%. Todas as amostras positivas por alt-Gen foram também positivas por nPCR. Ao comparar alt-Gen com qPCR, a sensibilidade foi de 100% para P. vivax, P. malariae e P. falciparum. Para todas as espécies Plasmodium, a correlação entre os valores limiares de ciclo de alt-S&T e alt-Gen comparados com qPCR foi significativa (P < 0,0001, teste de Spearman), com r = 0,8621 para alt-S&T e r = 0,9371 para alt-Gen. Quando todas as espécies de Plasmodium foram consideradas, houve uma correlação negativa entre o nível de parasitemia e os valores limiares do ciclo de PCR em tempo real (P < 0,0001). Neste estudo, apenas 2 de 28 amostras de indivíduos assintomáticos foram positivas por exame da gota espessa; no entanto, todas estas 28 amostras foram positivas por alt-S&T. Conclusões. Os ensaios alt-Gen e alt-S&T são adequados para a detecção de infecções submicroscópicas para fins epidemiológicos distintos, tais como para utilização em inquéritos e laboratórios de referência e o rastreio em bancos de sangue, o que contribuirá para os esforços globais de eliminação da malária.

ABSTRACT Objective. To evaluate molecular tools to detect low-level parasitemia and the five species of Plasmodium that infect humans for use in control and elimination programs, and in reference laboratories. Methods. We evaluated 145 blood samples from patients who tested positive by nested polymerase chain reaction (nPCR), from asymptomatic individuals and from the WHO Global Malaria Programme/United Kingdom National External Quality Assessment Service. Samples were assayed using the genus-specific RealStar® Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) and the RealStar® Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). The results from the molecular tests were compared with those from quantitative PCR (qPCR), nPCR and thick blood smear. Results. The levels of parasitemia ranged from 1 to 518 000 parasites/µL, depending on the species. Compared with nPCR, alt-S&T had a sensitivity of 100%, except for identifying P. falciparum, for which the sensitivity was 93.94%. All samples positive by alt-Gen were also positive by nPCR. When comparing alt-Gen to qPCR, the sensitivity was 100% for P. vivax, P. malariae and P. falciparum. For all Plasmodium species, the correlation between cycle threshold values of alt-S&T and alt-Gen compared with qPCR was significant (P < 0.0001, Spearman’s test), with r = 0.8621 for alt-S&T and r = 0.9371 for alt-Gen. When all Plasmodium species were considered, there was a negative correlation between the level of parasitemia and real-time PCR cycle threshold values (P < 0.0001). In this study, only 2 of 28 samples from asymptomatic individuals were positive by thick blood smear; however, all 28 of these samples were positive by alt-S&T. Conclusions. The alt-Gen and alt-S&T assays are suitable for detecting submicroscopic infections for distinct epidemiological purposes, such as for use in surveys and reference laboratories, and screening in blood banks, which will contribute to global efforts to eliminate malaria.

RESUMEN Objetivo. Evaluar herramientas moleculares para detectar bajos niveles de parasitemia y las cinco especies de Plasmodium que infectan a los seres humanos, a fin de emplearlas en los programas de control y eliminación y en los laboratorios de referencia. Métodos. Se evaluaron 145 muestras de sangre de pacientes positivos por reacción en cadena de la polimerasa anidada (nPCR), de individuos asintomáticos y de muestras del Programa Mundial de Malaria de la Organización Mundial de la Salud/Servicio Nacional de Evaluación Externa de Calidad del Reino Unido. Las muestras se analizaron con el kit de PCR RealStar® Malaria 1.0 (alt-Gen; altona Diagnostics), específico para cada género, y con el kit de PCR RealStar® Malaria Screen & Type (alt-S&T; altona Diagnostics). Se compararon los resultados de las pruebas moleculares con los de la PCR cuantitativa (qPCR), la nPCR y el frotis de gota gruesa. Resultados. Los niveles de parasitemia oscilaron entre 1 y 518 000 parásitos/µl, según la especie. En comparación con la nPCR, la prueba alt-S&T tuvo una sensibilidad del 100%, excepto para la identificación de P. falciparum, para el cual la sensibilidad fue del 93,94%. Todas las muestras positivas por alt-Gen lo fueron también por nPCR. Al comparar alt-Gen con la qPCR, la sensibilidad fue del 100% para P. vivax, P. malariae y P. falciparum. Para todas las especies de Plasmodium, la correlación entre los valores del umbral de ciclo de alt-S&T y alt-Gen en comparación con la qPCR fue significativa (P < 0,0001, prueba de Spearman), con r = 0,8621 para alt-S&T y r = 0,9371 para alt-Gen. Cuando se consideraron todas las especies de Plasmodium hubo una correlación negativa entre el nivel de parasitemia y los valores de umbral de ciclo de PCR en tiempo real (P < 0,0001). En este estudio, solo 2 de las 28 muestras de individuos asintomáticos fueron positivas por frotis de gota gruesa; sin embargo, las 28 muestras fueron positivas por alt-S&T. Conclusiones. Los ensayos alt-Gen y alt-S&T son adecuados para detectar infecciones submicroscópicas con distintos fines epidemiológicos, como su uso en investigaciones y laboratorios de referencia y el cribado en bancos de sangre, lo que contribuirá a los esfuerzos mundiales para eliminar la malaria.

https://doi.org/10.26633/rpsp.2022.11

COMVC-19: A Program to protect healthcare workers’ mental health during the COVID-19 Pandemic. What we have learned
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Fukuti, Pedro

; Uchôa, Caroline Louise Mesquita

; Mazzoco, Marina Flaborea

; Cruz, Isabella D'Andrea Garcia da

; Echegaray, Mariana V.F.

; Humes, Eduardo de Castro

; Silveira, Júlia Belizário

; Santi, Talita Di

; Miguel, Euripedes Constantino

; Corchs, Felipe

; Fatori, Daniel

; Campello, Guilherme

; Oliveira, Gabriel M. de

; Argolo, Felipe C.

; Ferreira, Felipe de M.

; Machado, Gustavo

; Argeu, Adriana

; Oliveira, Graça Maria Ramos de

; Serafim, Antônio de Pádua

; Siqueira, Luciana de Lima

; Rossi, Luciane de

; Rios, Izabel Cristina

; Oliveira, Talita Rodrigues de

; Antoniazzi, Leilane C. K.

; Gagliotti, Daniel Augusto Mori

; Abelama Neto, Emílio

; Oliveira Junior, Paulo Novais de

; Correia, Aline Villalobo

; Gonçalves, Luca Schilling

; Tortato, Liana Silva

; Busato, Wagner Machado Moraes

; Guimarães-Fernandes, Flávio

; Alves, Marcos

; Leite Netto, Oswaldo Ferreira

; Schoueri, Patrícia de Campos Lindenberg

; Roque, Márcio de Assis

; Merlin, Silvia Stahl

; Boer, Giovana Cardoso Machado

; Sallet, Paulo Clemente

; Malbergier, André

; Spedo, Mariana Abrahão

; Kamitsuji, Carla Satie

; Faria, Elizabeth de

; Moreira, Moacyr Vergara de Godoy

; Kaufman, Arthur

; Abdo, Carmita

; Scanavino, Marco de Tubino

; Lancman, Selma

; Tavares, Hermano

; Polanczyk, Guilherme

; Brunoni, André R.

; Forlenza, Orestes V.

; Barros-Filho, Tarcísio Eloy Pessoa de

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2021, Volume 76 elocation e2631

Abstract: En | Text: En | PDF: En

OBJECTIVE: In 2020, the COVID-19 pandemic brought a work and stress overload to healthcare workers, increasing their vulnerability to mental health impairments. In response, the authors created the COMVC-19 program. The program offered preventive actions and mental health treatment for the 22,000 workers of The Hospital das Clinicas da Faculdade de Medicina da Universidade de São Paulo (HCFMUSP). This paper aims to describe its implementation and share what we have learned from this experience. METHODS: Workers were able to easily access the program through a 24/7 hotline. Additionally, a mobile phone app that screened for signs and symptoms of emotional distress and offered psychoeducation and/or referral to treatment was made available. Data from both these sources as well as any subsequent psychiatric evaluations were collected. RESULTS: The first 20 weeks of our project revealed that most participants were female, and part of the nursing staff working directly with COVID-19 patients. The most frequently reported symptoms were: anxiety, depression and sleep disturbances. The most common diagnoses were Adjustment, Anxiety, and Mood disorders. CONCLUSIONS: Implementing a mental health program in a multimodal intervention was feasible in a major quaternary public hospital. Our data also suggests that preventive actions should primarily be aimed at anxiety and depression symptoms, with a particular focus on the nursing staff.

https://doi.org/10.6061/clinics/2021/e2631

The hidden Plasmodium malariae in blood donors: a risk coming from areas of low transmission of malaria
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Aschar, Mariana

; Levi, José Eduardo

; Farinas, Maria L. R. N.

; Montebello, Sandra C.

; Mendrone-Junior, Alfredo

; Di Santi, Silvia Maria

Revista do Instituto de Medicina Tropical de São Paulo

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2020, Volume 62 elocation e100

Abstract: En | Text: En | PDF: En

ABSTRACT Malaria is an infectious vector-borne disease with other important routes of transmission, such as blood transfusion and organ/tissue transplantation, due to asymptomatic reservoirs of Plasmodium presenting with low parasitemia. Reports of transfusion-transmitted malaria have shown that in immunosuppressed recipients, infections can be fatal if they are not diagnosed and timely treated. All Plasmodium species can survive on blood components at temperatures from 2 to 6 °C for some days or even weeks. This report describes two candidates for blood donation harboring Plasmodium, infected in an area considered non-endemic. Blood samples were collected from donors who attended a blood bank in Sao Paulo and tested by microscopy, qPCR for Plasmodium genus-specific amplification, targeting the parasite 18S ribosomal subunit gene and a multiplex qPCR based on mtDNA of the five species. Under microscopy, only structures resembling Plasmodium were observed. The qPCR whose standard curve tested parasites varying from 2 to 0.1 parasites/ µL, showed the presence of Plasmodium DNA in the two blood donors, as did the multiplex qPCR that revealed the presence of P. malariae. The prevalence of positive donors varies according to the level of transmission, ranging from 0.7 to 55% in endemic areas. In non-endemic regions, prevalences are lower, however, transfusion malaria can evolve to severe cases, due to the lack of suspicion of this transmission route. Asymptomatic donors from low transmission regions pose a risk to blood banks, with particular emphasis on those located in areas with malaria elimination goals.

https://doi.org/10.1590/s1678-9946202062100 359 downloads

Serological and molecular techniques applied for identification of Plasmodium spp. in blood samples from nonhuman primates
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Figueiredo, Mayra Araguaia Pereira

; Di Santi, Silvia Maria

; Manrique, Wilson Gómez

; André, Marcos Rogério

; Machado, Rosangela Zacarias

Revista Brasileira de Parasitologia Veterinária

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sep 2018, Volume 27 N. 3 Pages 363 - 376

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Resumo Neste estudo objetivamos identificar Plasmodium spp. em amostras sangue de primatas não humanos (PNH) do estado do Maranhão, utilizando técnicas clássicas e alternativas para o exame da malária humana. Foram analisadas 161 amostras de sangue de PNH, sendo 141 de CETAS (cativeiro) e 20 de reserva particular (vida livre), utilizando microscopia, teste de diagnóstico rápido (RDT), imunofluorescência indireta (IFI) e técnicas moleculares (semi-nested PCR, PCR em tempo real quantitativo e LAMP). Dois métodos sorológicos (dot-ELISA e ELISA indireto) também foram padronizados com antígenos solúveis de roptrias de P. falciparum e P. berghei. Formas trofozoíticas de Plasmodium sp. foram identificadas em lâminas de cinco animais diferentes. Nenhuma amostra foi positiva em TDR e LAMP. Quatro amostras foram soropositivas para P. malariae na IFI. Os soros de PNH mostraram baixa reatividade pelo ELISA indireto. Plasmodium sp. foi detectado em 34,16% (55/161) das amostras utilizando a qPCR baseada no gene 18S rRNA. No sequenciamento, duas amostras mostraram identidade com P. malariae (100%), uma com Plasmodium sp. ZOOBH (97%) e uma com P. falciparum (99%). A PCR mostrou ser a técnica mais sensível para diagnósticos de Plasmodium em amostras de PNH.

Abstract The aim of this study was to identify Plasmodium spp. in blood samples from nonhuman primates (NHPs) in the state of Maranhão, using classical and alternative techniques for examination of human malaria. A total of 161 blood samples from NHPs were analyzed: 141 from captive animals at a Wildlife Screening Center (CETAS) and 20 from free-living animals in a private reserve. The techniques used were microscopy, rapid diagnostic test (RDT), Indirect fluorescent antibody test (IFAT) and molecular techniques (semi-nested PCR, quantitative real-time PCR and LAMP). Two serological methods (dot-ELISA and indirect ELISA) were also standardized with rhoptry protein-soluble antigen of P. falciparum and P. berghei. Trophozoite forms of Plasmodium sp. were identified on slides from five different animals. No samples were positive through RDT and LAMP. Four samples were seropositive for P. malariae through IFAT. The samples showed low reactivity to ELISA. Plasmodium sp. was detected in 34.16% (55/161) of the samples using qPCR based on the 18S rRNA gene. After sequencing, two samples showed 100% identityl to P. malariae, one showed 97% identity to Plasmodium sp. ZOOBH and one showed 99% identity to P. falciparum . PCR was shown to be the most sensitive technique for diagnosing Plasmodium in NHP samples.

https://doi.org/10.1590/s1984-296120180043 1508 downloads

Natural Plasmodium infection in neotropical primates in the island of São Luís, state of Maranhão, Brazil
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Figueiredo, Mayra Araguaia Pereira

; Di Santi, Silvia Maria Fátima

; Figueiredo, Thaysa Araguaia Pereira

; Machado, Rosangela Zacarias

Revista Brasileira de Parasitologia Veterinária

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jun 2015, Volume 24 N. 2 Pages 122 - 128

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Os Estados que compõem a Amazônia Legal, entre eles o Estado do Maranhão, respondem a 99% dos casos registrados de malária humana no Brasil. Também se acredita que nessa região ocorra a transmissão de malária de primatas não humanos (PNH) para humanos, devido a relatos atuais de resultados soroepidemiológicos de amostras de humanos e PNH que coexistem nas mesmas áreas. O presente estudo objetivou realizar o diagnóstico morfológico, sorológico e molecular de Plasmodium spp. em primatas neotropicais na Ilha de São Luís, Estado do Maranhão, Brasil. Foram utilizadas como técnicas de diagnóstico: a microscopia de luz, a reação em cadeia pela polimerase (PCR) e a imunofluorescência indireta (RIFI). No período de junho de 2009 a abril de 2010, foram amostrados 70 PNH, sendo 50 provenientes do Centro de Triagem de Animais Silvestres (CETAS), localizado no município de São Luís, e 20 de vida livre, capturados em reserva particular localizada no município de São José de Ribamar, Estado do Maranhão. Foram avaliadas pela microscopia de luz 140 lâminas (duas de cada animal), das quais cinco animais (7,1%) foram positivos. Pela RIFI não se detectou anticorpos anti-Plasmodium spp. Pela PCR, dos 70 animais amostrados, foi possível observar produtos amplificados para Plasmodium spp. em 13 amostras, das quais oito (61,5%) eram de animais de vida livre e cinco (38,5%) de CETAS.

The states that make up the Legal Amazon Region, which include the state of Maranhão, account for 99% of registered cases of human malaria in Brazil. It is also believed that transmission of malaria from nonhuman primates (NHP) to humans occurs in this region, because of current reports of seroepidemiological results from samples from humans and NHP coexisting in the same areas. This study aimed to make morphological, serological and molecular diagnoses of Plasmodium spp. in neotropical primates on the island of São Luís, state of Maranhão, Brazil. The diagnostic techniques used were optical microscopy, the polymerase chain reaction (PCR) and the indirect immunofluorescence assay (IFA). From June 2009 to April 2010, 70 NHP were sampled: 50 at the Wild Animal Screening Center (CETAS), located in the municipality of São Luís and 20 free-living individuals that were caught in a private reserve located in the municipality of São Jose de Ribamar, state of Maranhão. Under an optical microscope, 140 slides (two from each animal) were evaluated and five animals (7.1%) were found to be positive. IFA did not detect anti-Plasmodium spp. From PCR on the 70 animals sampled, amplified Plasmodium spp. products were observed in 13 samples, of which eight (61.5%) were from free-living animals and five (38.5%) were from animals at CETAS.

https://doi.org/10.1590/S1984-29612015034 31239 downloads

Malaria in pregnant women living in areas of low transmission on the southeast Brazilian Coast: molecular diagnosis and humoural immunity profile
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Hristov, Angélica Domingues

; Sanchez, Maria Carmen Arroyo

; Ferreira, José Jarbas Bittencourt

; Lima, Giselle Fernandes Maciel de Castro

; Inoue, Juliana

; Costa-Nascimento, Maria de Jesus

; Sanchez, Arianni Rondelli

; Ramos-Sanchez, Eduardo Milton

; Santi, Silvia Maria Di

Memórias do Instituto Oswaldo Cruz

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dec 2014, Volume 109 N. 8 Pages 1014 - 1020

Abstract: En | Text: En | PDF: En

Studies on autochthonous malaria in low-transmission areas in Brazil have acquired epidemiological relevance because they suggest continued transmission in what remains of the Atlantic Forest. In the southeastern portion of the state of São Paulo, outbreaks in the municipality of Juquitiba have been the focus of studies on the prevalence of Plasmodium, including asymptomatic cases. Data on the occurrence of the disease or the presence of antiplasmodial antibodies in pregnant women from this region have not previously been described. Although Plasmodium falciparum in pregnant women has been widely addressed in the literature, the interaction of Plasmodium vivax and Plasmodium malariae with this cohort has been poorly explored to date. We monitored the circulation of Plasmodium in pregnant women in health facilities located in Juquitiba using thick blood film and molecular protocols, as well as immunological assays, to evaluate humoural immune parameters. Through real-time and nested polymerase chain reaction, P. vivax and P. malariae were detected for the first time in pregnant women, with a positivity of 5.6%. Immunoassays revealed the presence of IgG antibodies: 44% for ELISA-Pv, 38.4% for SD-Bioline-Pv and 18.4% for indirect immunofluorescence assay-Pm. The high prevalence of antibodies showed significant exposure of this population to Plasmodium. In regions with similar profiles, testing for a malaria diagnosis might be indicated in prenatal care.

https://doi.org/10.1590/0074-0276140229 1715 downloads

Malaria in Brazil: what happens outside the Amazonian endemic region
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Pina-Costa, Anielle de

; Brasil, Patrícia

; Santi, Sílvia Maria Di

; Araujo, Mariana Pereira de

; Suárez-Mutis, Martha Cecilia

; Santelli, Ana Carolina Faria e Silva

; Oliveira-Ferreira, Joseli

; Lourenço-de-Oliveira, Ricardo

; Daniel-Ribeiro, Cláudio Tadeu

Memórias do Instituto Oswaldo Cruz

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aug 2014, Volume 109 N. 5 Pages 618 - 633

Abstract: En | Text: En | PDF: En

Brazil, a country of continental proportions, presents three profiles of malaria transmission. The first and most important numerically, occurs inside the Amazon. The Amazon accounts for approximately 60% of the nation’s territory and approximately 13% of the Brazilian population. This region hosts 99.5% of the nation’s malaria cases, which are predominantly caused by Plasmodium vivax (i.e., 82% of cases in 2013). The second involves imported malaria, which corresponds to malaria cases acquired outside the region where the individuals live or the diagnosis was made. These cases are imported from endemic regions of Brazil (i.e., the Amazon) or from other countries in South and Central America, Africa and Asia. Imported malaria comprised 89% of the cases found outside the area of active transmission in Brazil in 2013. These cases highlight an important question with respect to both therapeutic and epidemiological issues because patients, especially those with falciparum malaria, arriving in a region where the health professionals may not have experience with the clinical manifestations of malaria and its diagnosis could suffer dramatic consequences associated with a potential delay in treatment. Additionally, because the Anopheles vectors exist in most of the country, even a single case of malaria, if not diagnosed and treated immediately, may result in introduced cases, causing outbreaks and even introducing or reintroducing the disease to a non-endemic, receptive region. Cases introduced outside the Amazon usually occur in areas in which malaria was formerly endemic and are transmitted by competent vectors belonging to the subgenus Nyssorhynchus (i.e., Anopheles darlingi, Anopheles aquasalis and species of the Albitarsis complex). The third type of transmission accounts for only 0.05% of all cases and is caused by autochthonous malaria in the Atlantic Forest, located primarily along the southeastern Atlantic Coast. They are caused by parasites that seem to be (or to be very close to) P. vivax and, in a less extent, by Plasmodium malariae and it is transmitted by the bromeliad mosquito Anopheles (Kerteszia) cruzii. This paper deals mainly with the two profiles of malaria found outside the Amazon: the imported and ensuing introduced cases and the autochthonous cases. We also provide an update regarding the situation in Brazil and the Brazilian endemic Amazon.

https://doi.org/10.1590/0074-0276140228 7263 downloads Cited 6 times in SciELO

Asymptomatic infection in individuals from the municipality of Barcelos (Brazilian Amazon) is not associated with the anti-Plasmodium falciparum glycosylphosphatidylinositol antibody response
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Gomes, Larissa Rodrigues

; Totino, Paulo Renato Rivas

; Sanchez, Maria Carmen Arroyo

; Daniel, Elsa Paula da Silva Kaingona

; Macedo, Cristiana Santos de

; Fortes, Filomeno

; Coura, Jose Rodrigues

; Santi, Silvia Maria Di

; Werneck, Guilherme Loureiro

; Suarez-Mutis, Martha Cecilia

; Ferreira-da-Cruz, Maria de Fatima

; Daniel-Ribeiro, Claudio Tadeu

Memórias do Instituto Oswaldo Cruz

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sep 2013, Volume 108 N. 6 Pages 796 - 800

Abstract: En | Text: En | PDF: En

Anti-glycosylphosphatidylinositol (GPI) antibodies (Abs) may reflect and mediate, at least partially, anti-disease immunity in malaria by neutralising the toxic effect of parasitic GPI. Thus, we assessed the anti-GPI Ab response in asymptomatic individuals living in an area of the Brazilian Amazon that has a high level of malaria transmission. For comparative purposes, we also investigated the Ab response to a crude extract prepared from Plasmodium falciparum, the merozoite surface protein (MSP)3 antigen of P. falciparum and the MSP 1 antigen of Plasmodium vivax (PvMSP1-19) in these individuals and in Angolan patients with acute malaria. Our data suggest that the Ab response against P. falciparum GPI is not associated with P. falciparum asymptomatic infection in individuals who have been chronically exposed to malaria in the Brazilian Amazon. However, this Ab response could be related to ongoing parasitaemia (as was previously shown) in the Angolan patients. In addition, our data show that PvMSP1-19may be a good marker antigen to reflect previous exposure to Plasmodium in areas that have a high transmission rate of P. vivax.

https://doi.org/10.1590/0074-0276108062013018 2037 downloads

10.

Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale
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Lima, Giselle FMC

; Levi, José E

; Geraldi, Marcelo P

; Sanchez, Maria Carmen A

; Segurado, Aluísio AC

; Hristov, Angélica D

; Inoue, Juliana

; Costa-Nascimento, Maria de Jesus

; Di Santi, Silvia M

Memórias do Instituto Oswaldo Cruz

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sep 2011, Volume 106 N. 6 Pages 691 - 700

Abstract: En | Text: En | PDF: En

Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.

11989 downloads

11.

Transfusion-transmitted malaria: case report of asymptomatic donor harboring Plasmodium malariae
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Scuracchio, Patricia

; Vieira, Sergio Domingos

; Dourado, Denise Albuquerque

; Bueno, Luciana Moro

; Colella, Rafael

; Ramos-Sanchez, Eduardo Milton

; Lima, Giselle F. M. Castro

; Inoue, Juliana

; Sanchez, Maria Carmen Arroyo

; Di Santi, Silvia Maria

Revista do Instituto de Medicina Tropical de São Paulo

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feb 2011, Volume 53 N. 1 Pages 55 - 59

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

No Brasil a malária é endêmica na Amazônia, porém casos autóctones com baixas parasitemias ocorrem na área costeira de Mata Atlântica. De acordo com a legislação brasileira, não são obrigatórios testes para detecção de malária em doadores de sangue de áreas não-endêmicas; entretanto são excluídos por seis meses aqueles com relato de deslocamento para áreas de transmissão. Este trabalho descreve um caso de malária transfusional ocorrido em São Paulo, Brasil, em que um paciente recebeu sangue infectado, desenvolvendo a doença. Ele residia em São Paulo e não apresentava histórico de transfusão anterior ou deslocamentos para áreas endêmicas, incluindo as de baixa endemicidade, como a Mata Atlântica. A gota espessa revelou Plasmodium malariae. Os doadores eram residentes em São Paulo e um deles (045-0) apresentou hemoscopia e PCR positivos. Este era assintomático com PCR positiva para P. malariae e viagem para Juquiá, Mata Atlântica de São Paulo, onde são descritos casos esporádicos de malária autóctone. Amostras de soro do receptor e de todos os doadores foram ensaiadas por ELISA com antígenos de P. vivax e P. falciparum e RIFI com P. malariae. O doador 045-0 apresentou RIFI positiva para P. malariae. ELISA-P. vivax foi reagente no doador infectado (045-0) e em dois dos 44 doadores. Todos os soros foram negativos com antígeno de P. falciparum. Este caso aponta a necessidade de revisão dos critérios de triagem clínico-epidemiológica para evitar casos transfusionais e também adequar as estratégias de exclusão de doadores de sangue.

Malaria in Brazil is endemic in the Amazon region, but autochthonous cases with low parasitaemia occur in the Atlantic Forest area of the country. According to Brazilian legislation no test is mandatory for blood donors from non-endemic areas. However if they have traveled to malaria transmission regions they are deferred for six months before they can donate. This report describes a transfusion-transmitted malaria case in Sao Paulo, Brazil, where one recipient received infected blood and developed the disease. He lived in Sao Paulo and had no previous transfusion or trips to endemic areas, including those of low endemicity, such as Atlantic Forest. Thick blood smears confirmed Plasmodiummalariae. All donors lived in Sao Paulo and one of them (Donor 045-0) showed positive hemoscopy and PCR. This asymptomatic donor had traveled to Juquia, in the Atlantic Forest area of S ao Paulo State, where sporadic cases of autochthonous malaria are described. DNA assay revealed P. malariae in the donor's (Donor 045-0) blood. Serum archives of the recipient and of all blood donors were analyzed by ELISA using both P. vivax and P. falciparum antigens, and IFAT with P. malariae. Donor 045-0's serum was P. malariae IFAT positive and the P. vivax ELISA was reactive. In addition, two out of 44 donors' archive sera were also P. vivax ELISA reactive. All sera were P. falciparum ELISA negative. This case suggests the need of reviewing donor selection criteria and deferral strategies to prevent possible cases of transfusion-transmitted malaria.

6980 downloads

12.

The monitoring of hematopoietic stem cell transplant donors and recipients from endemic areas for malaria
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Inoue, Juliana

; Machado, Clarisse Martins

; Lima, Giselle Fernandes Maciel de Castro

; Nascimento, Maria de Jesus Costa

; Colturato, Vergílio Rensi

; Di Santi, Silvia Maria

Revista do Instituto de Medicina Tropical de São Paulo

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oct 2010, Volume 52 N. 5 Pages 281 - 284

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

A malária é complicação incomum após o transplante de células-tronco hematopoiéticas em países endêmicos. No entanto, candidatos a transplantes, receptores e doadores que vivem em regiões endêmicas frequentemente relatam episódios anteriores de malária. Este fato pode representar um risco importante para receptores imunossuprimidos, que podem desenvolver casos de malária grave. Relatamos um caso de transplante de células-tronco hematopoiéticas (TCTH) em que o doador teve história de malária anterior e um monitoramento por meio de exames parasitológicos e moleculares foi realizado antes e após o procedimento. O doador apresentou Plasmodium vivax na gota espessa um mês após o transplante e foi tratado de acordo com as orientações do Ministério da Saúde brasileiro. A reação em cadeia da polimerase (PCR) foi capaz de detectar a infecção por malária no doador uma semana mais cedo do que a gota espessa. Mesmo sem resultados positivos, o receptor foi preventivamente tratado com cloroquina, a fim de prevenir as formas sanguíneas assexuadas. Destacamos a importância do monitoramento de receptores e doadores em procedimentos de transplante, com o objetivo de reduzir o risco de transmissão da malária.

Malaria is an unusual complication after hematopoietic stem cell transplantation in non-endemic countries. However, transplant candidates, recipients and donors living in endemic regions frequently report previous episodes of malaria. This fact could represent an important risk for immunosuppressed recipients that could develop severe malaria cases. We report a case of hematopoietic stem cell transplant (HSCT) in which the donor had a history of previous malaria, and close monitoring was performed before and after procedure by parasitological and molecular tests. The donor presented Plasmodium vivax in thick blood smears one month after transplant and was treated according to Brazilian Health Ministry guidelines. The polymerase chain reaction (PCR) was able to detect malaria infection in the donor one week earlier than thick blood film. Even without positive results, the recipient was pre-emptively treated with chloroquine in order to prevent the disease. We highlight the importance of monitoring recipients and donors in transplant procedures with the aim of reducing the risk of malaria transmission.

3006 downloads

13.

Malária autóctone notificada no Estado de São Paulo: aspectos clínicos e epidemiológicos de 1980 a 2007
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Couto, Renata D'Avila

; Latorre, Maria do Rosário Dias de Oliveira

; Di Santi, Silvia Maria

; Natal, Delsio

Revista da Sociedade Brasileira de Medicina Tropical

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feb 2010, Volume 43 N. 1 Pages 52 - 58

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

INTRODUÇÃO: A malária autóctone no Estado de São Paulo caracteriza-se por surtos esporádicos na região oeste e transmissão persistente na região leste onde ocorrem casos oligossintomáticos com baixa parasitemia pelo Plasmodium vivax. Os objetivos deste estudo foram: analisar a completitude das fichas de notificação de malária autóctone; estimar a tendência da incidência de casos autóctones no ESP de 1980 a 2007; analisar o comportamento clínico e epidemiológico dos casos em duas regiões de autoctonia neste período. MÉTODOS: Foi realizado um estudo descritivo com 18 variáveis das FIN de malária do ESP, analisadas em duas regiões e em dois períodos (1980-1993 e 1994-2007). Fontes de dados: SUCEN/SES/SP, SINAN/CVE/SES/SP e DATASUS. RESULTADOS: A completitude foi superior a 85% em 11 variáveis. A tendência da incidência foi decrescente. Foram notificados 821 casos autóctones, 91,6% na região leste, predominando Plasmodium vivax. A infecção assintomática teve maior porcentagem no segundo período (p<0,001). CONCLUSÕES: A completitude das informações foi considerada satisfatória. As diferenças clínicas encontradas merecem atenção da vigilância epidemiológica que deve lidar com o desafio da infecção assintomática por Plasmodium.

INTRODUCTION: Autochthonous malaria in the State of São Paulo is characterized by sporadic outbreaks in the western region and by persistent transmission in the eastern region, where oligosymptomatic cases with low parasitemia due to Plasmodium vivax occur. The objectives of this study were to assess the completeness of autochthonous malaria notification forms; to estimate the incidence trends of autochthonous cases in the State of São Paulo from 1980 to 2007; and to analyze the clinical and epidemiological patterns of cases in two autochthonous regions over this period. METHODS: This was a descriptive study that analyzed 18 variables on the notification form for malaria in the State of São Paulo, comparing these two regions over two periods (1980-1993 and 1994-2007). The data sources were SUCEN/SES/SP, SINAN/CVE/SES/SP and DATASUS. RESULTS: The completeness was greater than 85% for 11 variables. The incidence trend was decreasing. There were 821 autochthonous cases: 91.6% occurred in the eastern region, predominantly due to Plasmodium vivax. Asymptomatic infection had a higher percentage in the second period (p < 0.001). CONCLUSIONS: The completeness of the information was satisfactory. The clinical differences observed deserve attention from epidemiological surveillance agencies, which need to deal with the challenge of asymptomatic infection by Plasmodium.

5912 downloads Cited 4 times in SciELO

14.

PCR - based diagnosis to evaluate the performance of malaria reference centers
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Di Santi, Silvia Maria

; Kirchgatter, Karin

; Brunialti, Karen Cristina Sant'Anna

; Oliveira, Alessandra Mota

; Ferreira, Sergio Roberto Santos

; Boulos, Marcos

Revista do Instituto de Medicina Tropical de São Paulo

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aug 2004, Volume 46 N. 4 Pages 183 - 187

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Embora a gota espessa corada por Giemsa (GTS) permaneça o padrão ouro para o diagnóstico de malária, métodos moleculares são mais sensíveis e específicos para detectar parasitas e podem ser utilizados em centros de referência para avaliar o desempenho da microscopia. A descrição das seqüências dos genes ssrRNA de Plasmodium falciparum, P. vivax, P. malariae e P. ovale permitiu o desenvolvimento de uma reação em cadeia da polimerase (PCR) que tem sido utilizada para diferenciar as quatro espécies. O objetivo deste estudo foi determinar as espécies de Plasmodium através de PCR em 190 lâminas positivas de pacientes para verificar a qualidade do diagnóstico realizado no Laboratório de Malária da SUCEN. Considerando somente os 131 resultados positivos em ambas as técnicas, GTS detectou 4,6% de infecções mistas e 3,1% de P. malariae enquanto o PCR identificou 19,1% e 13,8%, respectivamente.

Although the Giemsa-stained thick blood smear (GTS) remains the gold standard for the diagnosis of malaria, molecular methods are more sensitive and specific to detect parasites and can be used at reference centers to evaluate the performance of microscopy. The description of the Plasmodium falciparum, P. vivax, P. malariae and P. ovale ssrRNA gene sequences allowed the development of a polymerase chain reaction (PCR) that had been used to differentiate the four species. The objective of this study was to determine Plasmodium species through PCR in 190 positive smears from patients in order to verify the quality of diagnosis at SUCEN's Malaria Laboratory. Considering only the 131 positive results in both techniques, GTS detected 4.6% of mixed and 3.1% of P. malariae infections whereas PCR identified 19.1% and 13.8%, respectively.

4358 downloads

15.

Evaluation of a rapid dipstick test, Malar-CheckTM, for the diagnosis of Plasmodium falciparum malaria in Brazil
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AVILA, Priscilla Elisangela

; KIRCHGATTER, Karin

; BRUNIALTI, Karen Cristina S.

; OLIVEIRA, Alessandra M.

; SICILIANO, Rinaldo F.

; DI SANTI, Silvia Maria

Revista do Instituto de Medicina Tropical de São Paulo

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oct 2002, Volume 44 N. 5 Pages 293 - 296

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Este trabalho avaliou o Malar-CheckTM Pf test, ensaio imunocromatográfico que detecta a proteína rica em histidina de Plasmodium falciparum, dispensa uso de equipamentos, é rápido e de fácil execução. Ensaios de diluição com o isolado Palo Alto ou sangue de pacientes com P. falciparum, foram realizados para testar a sensibilidade em diferentes densidades do parasita. Malar-CheckTM foi comparado à gota espessa (GE), padrão ouro para diagnóstico de malária. A média do limiar de sensibilidade para cada técnica em três experimentos independentes foi de 12 e 71 parasitas/mm³ (GE e Malar-CheckTM, respectivamente). Em ensaios de campo, amostras foram coletadas de pacientes febris de áreas endêmicas. Comparado à GE, Malar-CheckTM foi verdadeiramente positivo em 38 pacientes, falso positivo em 3, verdadeiramente negativo em 23 e falso negativo em um. Malar-CheckTMrealizado com sangue de pacientes com P. falciparum após tratamento mostrou persistência do antígeno durante 30 dias. Malar-CheckTM pode ser útil no diagnóstico de P. falciparum em áreas remotas e auxiliar a rotina diagnóstica, mesmo quando a microscopia está disponível. Deve ser considerada infecção pregressa por P. falciparum, que pode determinar testes positivos em indivíduos curados. A rapidez do Malar-CheckTM para o diagnóstico precoce pode evitar evolução para casos graves.

The present study was carried out to evaluate the Malar-CheckTM Pf test, an immunochromatographic assay that detects Plasmodium falciparum Histidine Rich Protein II, does not require equipment, and is easy and rapid to perform. In dilution assays performed to test sensitivity against known parasite density, Malar-CheckTMwere compared with thick blood smear (TBS), the gold standard for diagnosis. Palo Alto isolate or P. falciparum blood from patients with different parasitemias was used. The average cut-off points for each technique in three independent experiments were 12 and 71 parasites/mm³ (TBS and Malar-CheckTM, respectively). In the field assays, samples were collected from patients with fever who visited endemic regions. Compared to TBS, Malar-CheckTMyielded true-positive results in 38 patients, false-positive results in 3, true-negative results in 23, and false-negative result in 1. Malar-CheckTMperformed with samples from falciparum-infected patients after treatment showed persistence of antigen up to 30 days. Malar-CheckTM should aid the diagnosis of P. falciparum in remote areas and improve routine diagnosis even when microscopy is available. Previous P. falciparum infection, which can determine a false-positive test in cured individuals, should be considered. The prompt results obtained with the Malar-CheckTM for early diagnosis could avoid disease evolution to severe cases.

4250 downloads Cited 2 times in SciELO

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