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Doctoral Thesis
DOI
https://doi.org/10.11606/T.17.2021.tde-11042022-111537
Document
Author
Full name
Felipe Santos de Carvalho
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2021
Supervisor
Committee
Figueiredo, Luiz Tadeu Moraes (President)
Angerami, Rodrigo Nogueira
Fonseca, Benedito Antonio Lopes da
Labruna, Marcelo Bahia
Title in Portuguese
Pesquisa de Rickettsia sp. em carrapatos e pacientes atendidos na região Ribeirão Preto, utilizando a PCR
Keywords in Portuguese
Diagnóstico
Febre maculosa
PCR
Rickettsia
Abstract in Portuguese
Rickettsioses são zoonoses transmitidas por artrópodos, especialmente carrapatos, de distribuição ubíqua, sendo cada rickettsiose caracterizada por aspectos clínicos-epidemiológicos associados à espécie envolvida e a ecologia de seus vetores. A Febre Maculosa Brasileira (FMB), principal rickettsiose no País, causada por Rickettsia rickettsii é considerada uma doença emergente e reemergente, que possui alta letalidade, apesar de ter boa resposta a antibioticoterapia de baixo custo. Estabelecer o diagnóstico precocemente é um desafio, pois não estão disponíveis testes diagnósticos rápidos e de alta acurácia. O presente estudo pesquisou DNA rickettsial por PCR, em pacientes e carrapatos na região de Ribeirão Preto/SP. Ao todo, foram analisadas 482 amostras, sendo 90 obtidas de 35 pacientes participantes; 255 amostras de banco de soro pacientes com síndrome febril aguda; 89 amostras de carrapatos representando 1633 animais, dos quais 978 coletados para o estudo (ninfas e larvas distribuídas em pools e adultos analisados individualmente), 654 de 19 pools previamente armazenados e um espécime encontrado em um paciente; e ainda 48 amostras de soro estocados de vertebrados. Em todas as amostras, foram usadas PCR's convencional em tempo real para o gene gltA. Os casos positivos foram avaliados por PCR's convencionais para fragmento maior de gltA, ompB e por heminested para ompA. 4,1% das amostras foram positivas na PCR em tempo real, sendo duas amostras de pacientes participantes, duas de soroteca e 16 amostras de carrapatos capturados. Nenhuma amostra foi positiva com as técnicas convencionais para gltA e ompB. Uma amostra teve amplificação gênica no protocolo hemi-nested para ompA. O presente estudo confirmou a presença de rickettsia na região de Ribeirão Preto/SP por PCR. Técnicas moleculares podem ser úteis no diagnóstico e compreensão epidemiológica das rickettsioses.
Title in English
Survey for Rickettsia sp. in ticks and patients attended in the region of Ribeirao Preto, using PCR
Keywords in English
Diagnosis
PCR
Rickettsia
Spotted fever
Abstract in English
Rickettsial diseases are zoonosis transmitted by arthropods, especially ticks. They have a ubiquitous distribution and each rickettsiosis has a characteristic clinical and epidemiological pattern associated with its own rickettsial species and the ecology of its specific vectors. Brazilian Spotted Fever (BSF), caused by Rickettsia rickettsii is the most important rickettsiosis in the country and is thought to be an emerging and reemerging disease. It is an acute febrile illness with high death rate, although it has a good response to low-cost antimicrobial therapy. One of the difficulties involving BSF is the lack of rapid diagnostic tests. A major challenge to physicians is to establish diagnosis as early as possible, allowing therapy to be started sooner, but fast and high accurate tests are not available. This study searched for rickettsial DNA using PCR in patients and ticks in the Ribeirão Preto region, state of São Paulo. 482 samples were analyzed: 90 samples from 35 patients enrolled in the study; 255 from sera bank of acute febrile patients; and 89 tick samples representing 1633 animals, of which 978 were collected for this study (nymphs and larvae in pools and adults individually), 654 from 19 pools previously stocked and one tick found feeding on a patient; and 48 serum samples previously obtained from wild vertebrate animals. Conventional and real time PCR targeting a gltA gene were performed in all samples. Positive cases were additionally analyzed by conventional PCRs for a long fragment of gltA, ompB and a hemi-nested protocol for ompA. 4.1% samples were positive by the real time PCR, two samples from enrolled patients, other two from sera bank and 16 samples from collected ticks. None of the samples were positive with conventional PCR for gltA or ompB genes. One sample were also positive with the hemi-nested PCR for ompA. This study has confirmed rickettsia in Ribeirão Preto region. Molecular methods can be useful for clinical diagnosis and epidemiological studies of rickettsiosis.
 
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Publishing Date
2022-04-13
 
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