Development of Real Time RT-PCR Techniques for arbovirus that cause human disease and nucleotide sequencing identification of them

• Grant number: 12/02836-6
• Support Opportunities: Scholarships in Brazil - Master
• Effective date (Start): November 01, 2012
• Effective date (End): August 31, 2014
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Luiz Tadeu Moraes Figueiredo
• Grantee: Marilia Farignoli Romeiro
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The arboviruses related families Flaviviridae, Togaviridae, and Bunyaviridae include serious public health problems in tropical and subtropical countries. These arboviruses generally misdiagnosed, concerning clinical manifestations ranging from asymptomatic infection, acute febrile disease viral hemorrhagic fever and encephalitis.These arboviruses are maintained in nature through cycles that comprise natural hosts (amplifiers) like birds, mammals and some reptiles, and in vectors of the genus Aedes sp. mosquitoes, Culex sp., Sabethes sp. and Psorophora sp., sometimes they can transmit these viruses to accidental hosts like humans, horses and other mammals. The methodologies used in the diagnosis of arboviral infection are based on serological tests such as ELISA, and virological tests. With the revolution of molecular techniques, the polymerase chain reaction (PCR) have been widely used in the diagnosis of arboviral infections. The technique of real time RT-PCR brings advantages over conventional PCR technique highlighting the high sensitivity and specificity, and quantification of viral load from samples. The present study aims to develop a real time RT-PCR based on SYBR Green detection system to diagnosis human arboviral diseases and investigates, by sequencing analisys, the presence of Flavivirus, Alphavirus, Phlebovirus and Orthobunyavirus in human samples.

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ROMEIRO, MARILIA FARIGNOLI; DE SOUZA, WILLIAM MARCIEL; TOLARDO, ALINE LAVADO; VIEIRA, LUIZ CARLOS; HENRIQUES, DYANA ALVES; DE ARAUJO, JANSEN; HASSEGAWA SIQUEIRA, CARLOS EDUARDO; COLOMBO, TATIANA ELIAS; AQUINO, VICTOR HUGO; LOPES DA FONSECA, BENEDITO ANTONIO; et al. A real-time RT-PCR for rapid detection and quantification of mosquito-borne alphaviruses. ARCHIVES OF VIROLOGY, v. 161, n. 11, p. 3171-3177, NOV 2016. (13/02256-2, 12/02836-6, 13/14929-1, 12/24150-9, 14/20851-8)

ALINE LAVADO TOLARDO; WILLIAM MARCIEL DE SOUZA; MARILIA FARIGNOLI ROMEIRO; LUIZ CARLOS VIEIRA; LUCIANO KLEBER DE SOUZA LUNA; DYANA ALVES HENRIQUES; JANSEN DE ARAUJO; CARLOS EDUARDO HASSEGAWA SIQUEIRA; TATIANA ELIAS COLOMBO; VICTOR HUGO AQUINO; et al. A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus. Memórias do Instituto Oswaldo Cruz, v. 111, n. 6, p. 385-390, 2016-06-03. (13/02256-2, 13/14929-1, 12/24150-9, 12/02836-6)

SILVA, JAQUELINE RAYMONDI; ROMEIRO, MARILIA FARIGNOLI; DE SOUZA, WILLIAM MARCIEL; MUNHOZ, THIAGO DEMARCHI; BORGES, GUSTAVO PUIA; BRIOSCHI SOARES, OTAVIO AUGUSTO; COELHO DE CAMPOS, CARLOS HENRIQUE; MACHADO, ROSANGELA ZACARIAS; CRISTINY RODRIGUES SILVA, MARIA LUANA; MAIA FARIA, JOICE LARA; et al. A Saint Louis encephalitis and Rocio virus serosurvey in Brazilian horses. Revista da Sociedade Brasileira de Medicina Tropical, v. 47, n. 4, p. 414-417, JUL-AUG 2014. (08/50617-6, 12/02836-6, 12/24150-9)

MOTTA MAIA, FELIPE GONCALVES; CHAVEZ, JULIANA HELENA; DE SOUZA, WILLIAM MARCIEL; ROMEIRO, MARILIA FARIGNOLI; DE CASTRO-JORGE, LUIZA ANTUNES; LOPES DA FONSECA, BENEDITO ANTONIO; MORAES FIGUEIREDO, LUIZ TADEU. Infection with Saint Louis encephalitis virus in the city of Ribeirao Preto, Brazil: report of one case. INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, v. 26, p. 96-97, SEP 2014. (08/50617-6, 12/02836-6, 12/24150-9)

ROMEIRO, MARILIA FARIGNOLI; DE SOUZA, WILLIAM MARCIEL; TOLARDO, ALINE LAVADO; VIEIRA, LUIZ CARLOS; COLOMBO, TATIANA ELIAS; AQUINO, VICTOR HUGO; NOGUEIRA, MAURICIO LACERDA; MORAES FIGUEIREDO, LUIZ TADEU. Evaluation and optimization of SYBR Green real-time reverse transcription polymerase chain reaction as a tool for diagnosis of the Flavivirus genus in Brazil. Revista da Sociedade Brasileira de Medicina Tropical, v. 49, n. 3, p. 279-285, MAY-JUN 2016. (13/02256-2, 12/02836-6, 14/20851-8, 12/24150-9, 13/14929-1)