In Brazil, the sensitivity was 83
In Brazil, the sensitivity was 83.6% for confirmed positive samples for DENV-1, DENV-2, and DENV-3, and better results were obtained using samples obtained up to the fifth day in the viremic phase [12]. for the emergence of severe cases [3], [4]. However, the greatest epidemic of dengue in Brazil occurred in 2002, when almost 800,000 cases of the disease were reported, most of them DENV-3 [5]. In 2010 2010, DENV-4 was detected in Boa Vista [6]. Since 2009, the NS1 antigen-capture ELISA test has been implemented by the Health Ministry in the Public Health laboratories network. Patients arriving for care within five days of the onset of disease symptoms are screened for viral isolation in health care sentinel Fenoprofen calcium models and, thus, provide early detection of circulating serotypes in a given area [7]. Fenoprofen calcium Open in a separate window Physique 1 Reverse transcriptase-semi-nested-polymerase chain reaction (RT-PCR) product in 2% agarose gel stained with syber safe.Lines 1C4: positive samples; 5C8: negative samples; 9C12: positive control for DENV-1, DENV-2, DENV-3 and DENV-4; NC: unfavorable control; MW: molecular excess weight marker. In Sergipe state, routine NS1 Ag test detection protocol is applied to patients in the sentinel models according to Health Ministry determinations. Blood is collected and send to the Central Laboratory (Laboratrio Central – LACEN/SE) for the Platelia? test. Positive biological samples are then forwarded to the reference laboratory in Rio de Janeiro for viral isolation. Unfavorable samples are stored in a ?80C and for this study were provided to our group in order to evaluate the presence of other arbovirus. Methods Ethics Statement This study followed the ethical principles according to the Helsinki Declaration and was approved by the Research Ethics Committee of the Federal University or college Fenoprofen calcium of Sergipe (02872312.7.0000.0058). The samples analyzed were derived from routine diagnosis for dengue fever conducted by the Central Laboratory of Public Health (LACEN). Patients were asked and provided their oral consent to enter in the Ministry Fenoprofen calcium of Health protocol Rabbit Polyclonal to Cyclin C (phospho-Ser275) of diagnosis and isolation of serotype dengue computer virus. They were all informed that this serum was kept for further investigations of the blood circulation of other arboviruses and they were asked to return after 14 days for serological diagnosis. As that step was part of the routine of the dengue control program, there was not written term of consent available. Ethics committee dispensed patients written informed consent after we offered signed agreement by the authority responsible for the custody of the samples. Confidentiality of the identity and results was managed. Study To investigate the co-circulation of arboviruses in the State of Sergipe, blood samples unfavorable for the NS1 antigen test from suspected dengue patients were requested from your Central Laboratory of Public Health. Platelia Dengue NS1 Ag test is usually a one-step sandwich-format microplate enzyme immunoassay for detecting dengue computer virus NS1 antigen. Assessments were carried out on samples of serum or blood from patients and controls according to the manufacturers recommendations. Before being included in the protocol for the detection of other arboviruses these unfavorable blood samples were initially evaluated by semi-nested RT-PCR for dengue [8] at the Evandro Chagas Institute (Instituto Evandro Chagas C IEC) to exclude possible dengue cases that might not have been detected by NS1 antigen-capture ELISA. In total, 119 samples of serum or blood from symptomatic patients collected between September 2011 and February 2012 were obtained, and sample RNA was extracted using TRIzol? Plus reagent (Invitrogen?, California, US) according to the manufacturers instructions. Target.