A Multiplex Nanopore Sequencing Approach for the Detection of Multiple Arboviral Species.

The emergence and continued geographic expansion of arboviruses and the growing number of infected people have highlighted the need to develop and improve multiplex methods for rapid and specific detection of pathogens. Sequencing technologies are promising tools that can help in the laboratory diagnosis of conditions that share common symptoms, such as pathologies caused by emerging arboviruses. In this study, we integrated nanopore sequencing and the advantages of reverse transcription polymerase chain reaction (RT-PCR) to develop a multiplex RT-PCR protocol for the detection of Chikungunya virus (CHIKV) and several orthoflaviviruses (such as dengue (Orthoflavivirus dengue), Zika (Orthoflavivirus zikaense), yellow fever (Orthoflavivirus flavi), and West Nile (Orthoflavivirus nilense) viruses) in a single reaction, which provides data for sequence-based differentiation of arbovirus lineages.

Cognitive Dysfunction of Chikungunya Virus Infection in Older Adults.

Background: Chikungunya fever is a disabling articular disease caused by chikungunya virus (CHIKV). In the past decade it has affected millions of people across America, Africa, Asia, and Europe, turning this infection into a public health concern. The acute phase of chikungunya infection is usually self-limiting, characterized by severe arthralgia, fever, chills, myalgia, headache, and rash. CHIKV neurovirulence is evident and seems to be higher among elders. Considering their susceptibility to cognitive decline and dementia, the aim of our study was to investigate whether CHIKV infection might cause long-term cognitive impairment in aged people. Methods: A cross-sectional study was conducted with volunteers aged from 60 to 90 who had been affected by chikungunya and also with healthy controls. A structured questionnaire was used to record demographic and clinical data, functional status, and depression. Global cognitive function was assessed through MoCA. A comprehensive neuropsychological battery was performed to assess specific cognitive functions. Results: Subjective memory complaints were present in 70% of subjects with previous chikungunya. This group had a poorer performance in MoCA (p = 0.000) and specific cognitive tests: Semantic (p = 0.05) and Phonemic Verbal Fluency (p = 0.003), 5-Digit (choice, reading, counting and alternance, p = 0.003, p = 0.014, p = 0.021, and p = 0.021, respectively), Stroop test (time, errors and interference, p = 0.000, p = 0.027 and p = 0.015, respectively), and RAVLT (word total session p = 0.05). These tests reflect performance on general executive functions, cognitive flexibility, inhibitory control, processing speed, semantic memory and episodic memory. Conclusion: Our data suggest that CHIKV infection may cause long-term cognitive decline in aged people and might be a risk factor for future dementia in this population.

Pathology of congenital Zika syndrome in Brazil: a case series.

BACKGROUND: Zika virus is an arthropod-borne virus that is a member of the family Flaviviridae transmitted mainly by mosquitoes of the genus Aedes. Although usually asymptomatic, infection can result in a mild and self-limiting illness characterised by fever, rash, arthralgia, and conjunctivitis. An increase in the number of children born with microcephaly was noted in 2015 in regions of Brazil with high transmission of Zika virus. More recently, evidence has been accumulating supporting a link between Zika virus and microcephaly. Here, we describe findings from three fatal cases and two spontaneous abortions associated with Zika virus infection. METHODS: In this case series, formalin-fixed paraffin-embedded tissue samples from five cases, including two newborn babies with microcephaly and severe arthrogryposis who died shortly after birth, one 2-month-old baby, and two placentas from spontaneous abortions, from Brazil were submitted to the Infectious Diseases Pathology Branch at the US Centers for Disease Control and Prevention (Atlanta, GA, USA) between December, 2015, and March, 2016. Specimens were assessed by histopathological examination, immunohistochemical assays using a mouse anti-Zika virus antibody, and RT-PCR assays targeting the NS5 and envelope genes. Amplicons of RT-PCR positive cases were sequenced for characterisation of strains. FINDINGS: Viral antigens were localised to glial cells and neurons and associated with microcalcifications in all three fatal cases with microcephaly. Antigens were also seen in chorionic villi of one of the first trimester placentas. Tissues from all five cases were positive for Zika virus RNA by RT-PCR, and sequence analyses showed highest identities with Zika virus strains isolated from Brazil during 2015. INTERPRETATION: These findings provide strong evidence of a link between Zika virus infection and different congenital central nervous system malformations, including microcephaly as well as arthrogryposis and spontaneous abortions. FUNDING: None.

Notes from the Field: Evidence of Zika Virus Infection in Brain and Placental Tissues from Two Congenitally Infected Newborns and Two Fetal Losses--Brazil, 2015.

Zika virus is a mosquito-borne flavivirus that is related to dengue virus and transmitted primarily by Aedes aegypti mosquitoes, with humans acting as the principal amplifying host during outbreaks. Zika virus was first reported in Brazil in May 2015 (1). By February 9, 2016, local transmission of infection had been reported in 26 countries or territories in the Americas.* Infection is usually asymptomatic, and, when symptoms are present, typically results in mild and self-limited illness with symptoms including fever, rash, arthralgia, and conjunctivitis. However, a surge in the number of children born with microcephaly was noted in regions of Brazil with a high prevalence of suspected Zika virus disease cases. More than 4,700 suspected cases of microcephaly were reported from mid-2015 through January 2016, although additional investigations might eventually result in a revised lower number (2). In response, the Brazil Ministry of Health established a task force to further investigate possible connections between the virus and brain anomalies in infants (3).

Reação leucemoide e anemia hemolítica grave causada por Mycoplasma pneumoniae / Leukemoid reaction and severe hemolytic anemia due to Mycoplasma pneumoniae

Hemólise massiva é uma manifestação rara da infecção por Mycoplasma pneumoniae.É desencadeada por crioaglutininas, anticorpos IgM, que surgem sete a dez dias após a infecção em cerca de 50 por cento a 75 por cento dos casos. Hiperleucocitose é também evento incomum e orienta o diagnóstico para etiologia bacteriana ou neoplásica. Relatamos um caso de um homem de 67 anos com pneumonia por Mycoplasma pneumoniae, que apresentou anemia grave e reação leucemoide, diagnosticada por meio do teste de crioaglutininas à beira do leito e dosagem do título de anticorpo anti-mycoplasma. Após início do tratamento adequado, houve melhora importante da anemia e a leucometria foi reduzida de 56.100/mm³ para valores próximos do normal.

Massive hemolysis, a rare manifestation of Mycoplasma pneumoniae infection, is due to cold agglutinins (IgM antibodies) that appear seven to ten days after the infection in around 50 percent to 70 percent of cases. Hyperleukocytosis, suggestive of bacterial or neoplastic etiologies, is also an uncommon event. We report here on the case of a 67-year-old man with Mycoplasma pneumoniae pneumonia who presented with severe anemia and leukemoid reaction as diagnosed by the bedside cold agglutinin test and measurement of anti-mycoplasma antibodies. After beginning appropriate treatment, the anemia improved significantly and the leukocyte count reduced from 56100 /mm³ to close to normal level.

Incidência pós-transfusional do HBsAg em crianças com doenças neoplásicas / Post-transfusional incidence of HBsAg in children with neoplastic diseases

Há 350 milhões de indivíduos cronicamente infectados pelo vírus da hepatite B (VHB). O desenvolvimento e a gravidade dessa infecção dependem de vários fatores, tais como a idade da primoinfecção e resposta imune, sendo o risco dessa cronificação menor que 5 por cento em adultos e maior que 90 por cento em neonatos. Indivíduo cronicamente infectado pelo VHB durante a infância tem 25 por cento de chance de morrer por cirrose ou hepatocarcinoma. Tais evidências conduziram os autores a desenvolver este estudo cujo objetivo foi estimar a incidência pós-transfusional do antígeno de superfície do vírus da hepatite B (HBsAg) em crianças com neoplasias, que foram transfundidas no curso do tratamento ou seguimento da doença. Foi realizado um estudo retrospectivo, com revisão de 333 prontuários do serviço de oncologia do HIVS, de janeiro de 1993 a janeiro de 2005. Adotaram-se como critérios de inclusão: idade menor que 16 anos, diagnóstico de doença neoplásica e realização da pesquisa do HBsAg. Assim, 199 prontuários foram excluídos por não preencherem tais critérios, restando 134 que foram analisados quanto à realização de hemotransfusão. Das 134 crianças, 116 foram transfundidas e 18 não. Apresentaram pesquisa reativa para o HBsAg 32,8 por cento das transfundidas e apenas 5,6 por cento das não-transfundidas. O teste Exato de Fisher mostrou que houve significância estatística (p = 0,023) e observou-se que as proporções de pacientes com pesquisa reativa diferem entre os transfundidos e os não-transfundidos. O odds ratio de um paciente transfundido apresentar pesquisa reativa para o HBsAg foi calculado em 8,28 vezes maior do que um não-transfundido.

There are 350 million individuals infected by the hepatitis B virus (HBV). The development and the severity of the infection depend on several factors, such as: age at the first infection and the immunity response of the subject. The risk of chronic infection is less than 5 percent for adults and greater than 90 percent for newborn babies. An individual who is chronically infected by HBV during childhood has a 25 percent risk of dying due to cirrhosis or liver cancer. This data led the authors to design a study with the objective of estimating the post-transfusion incidence of the hepatitis B virus surface antigen (HBsAg) in children with neoplasias who were transfused during treatment or during the follow up. A retrospective study was performed that revisited 333 medical records from the oncology service in the HIVS from January 1993 to January 2005. The inclusion criteria were: age less than 16 years old, diagnosis of cancer and the results of the HBsAg test. Thus, 199 patients were excluded because they did not fulfill the criteria. The remaining 134 patients' records were analyzed in regards to blood transfusion. Of the 134 children who satisfied the criteria, 116 were transfused and 18 were not. Results of the HBsAg test were positive in 32.8 percent of the transfused patients and in only 5.6 percent of non-transfused individuals. The Fisher Exact Test demonstrated a statistically significant difference (p = 0.023). The ODDS ratio of a transfused patient presenting with reactive results for HBsAG was calculated at 8.28 times greater than non-transfused individuals.