Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue

Detalhes bibliográficos
Autor(a) principal: Nakashima, Fabiana
Data de Publicação: 2015
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da FAMERP
Texto Completo: http://bdtd.famerp.br/handle/tede/267
Resumo: Introduction: Toxoplasma gondii, which causes toxoplasmosis, is an intracellular protozoan that has several transmission routes, among them the transfusion. Objective: To investigate by serological and molecular methods the T. gondii infection in blood donors to assess the risk of transmission via transfusion. Methods: We selected 750 individuals able to donate blood. These individuals were submitted to an interview about their lifestyle habits and a collection of peripheral blood. The immunosorbent assay (ELISA) was used to investigate the presence of anti-T. gondii IgM and IgG classes, and the Nested PCR and qQPCR, the parasitaemia. Positive samples in the molecular methods were submitted for genotyping by the Multilocus Nested PCR RFLP method. Donors with positive molecular result and no positive serology were recalled to investigate seroconversion by ELISA and Indirect Immunofluorescence (IIF). In addition, recipients of blood components with molecular suspected parasitemia were screened. Of these, aliquots were separated from peripheral blood prior to transfusion and after transfusion (± 20 days), which were also analyzed by serological methods (ELISA and IIF) and molecular (PCR, qPCR Mulitplex nested PCR and RFLP). Results: Three hundred and fifty-seven (47.6%) donors had positive result and 393 (52.4%) showed no positive result for the presence of anti-T. gondii IgG class. Twenty-seven (3,6%) showed positive result for IgM and 723 (96.4%) non-reactive. The variables associated with infection by T. gondii were: advanced age (P < 0.0001), consumption of raw milk (P = 0.001), consumption of raw and underare beef and pork meat (P = 0.003), to reside in the countryside (P = 0.004), frequent presence of mechanical vectors in the residence (cockroach, mouse and fly P = 0.02; mice, P = 0.03), lower education (1st grade incomplete, P <0.0001 and 1st grade complete, P = 0.002) and low family income (P = 0.002). No sample was positive by qPCR, but 40 (11%) were positive by Nested PCR and positive serology. However, these samples did not show amplification when undergoing to genotyping. The four cases screened did not show positive results to molecular analysis and no recalled donor presented seroconversion. Conclusion: We concluded that the studied population of blood donors is exposed to various risk factors associated with infection by T. gondii, and it is likely that the high frequency of anti-T. gondii IgG class found in this study is related to this exposure. The results show that there is no molecular evidence of parasitaemia in the studied donors, thus the risk of transmission of this infection via blood transfusion is low or zero in this region. In addition, the failure of genotyping and the non-occurrence of seroconversion of reanalyzed donors suggest that the positivity found in some samples analyzed by Nested PCR, were related to false-positive results.
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spelling Mattos, Luiz Carlos deMattos, Cinara de Cássia Brandão deCastiglioni, LilianRicci Junior, OctávioFerreira, Ana Iara da CostaMartin, Natália34254202830http://lattes.cnpq.br/9715943105309778Nakashima, Fabiana2016-06-20T21:04:05Z2015-12-14Nakashima, Fabiana. Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue. 2015. 118 p. Tese ( Programa de Pós-Graduação em Ciências da Saúde) - Faculdade de Medicina de São José do Rio Preto, São José do Rio Preto .1159http://bdtd.famerp.br/handle/tede/267Introduction: Toxoplasma gondii, which causes toxoplasmosis, is an intracellular protozoan that has several transmission routes, among them the transfusion. Objective: To investigate by serological and molecular methods the T. gondii infection in blood donors to assess the risk of transmission via transfusion. Methods: We selected 750 individuals able to donate blood. These individuals were submitted to an interview about their lifestyle habits and a collection of peripheral blood. The immunosorbent assay (ELISA) was used to investigate the presence of anti-T. gondii IgM and IgG classes, and the Nested PCR and qQPCR, the parasitaemia. Positive samples in the molecular methods were submitted for genotyping by the Multilocus Nested PCR RFLP method. Donors with positive molecular result and no positive serology were recalled to investigate seroconversion by ELISA and Indirect Immunofluorescence (IIF). In addition, recipients of blood components with molecular suspected parasitemia were screened. Of these, aliquots were separated from peripheral blood prior to transfusion and after transfusion (± 20 days), which were also analyzed by serological methods (ELISA and IIF) and molecular (PCR, qPCR Mulitplex nested PCR and RFLP). Results: Three hundred and fifty-seven (47.6%) donors had positive result and 393 (52.4%) showed no positive result for the presence of anti-T. gondii IgG class. Twenty-seven (3,6%) showed positive result for IgM and 723 (96.4%) non-reactive. The variables associated with infection by T. gondii were: advanced age (P < 0.0001), consumption of raw milk (P = 0.001), consumption of raw and underare beef and pork meat (P = 0.003), to reside in the countryside (P = 0.004), frequent presence of mechanical vectors in the residence (cockroach, mouse and fly P = 0.02; mice, P = 0.03), lower education (1st grade incomplete, P <0.0001 and 1st grade complete, P = 0.002) and low family income (P = 0.002). No sample was positive by qPCR, but 40 (11%) were positive by Nested PCR and positive serology. However, these samples did not show amplification when undergoing to genotyping. The four cases screened did not show positive results to molecular analysis and no recalled donor presented seroconversion. Conclusion: We concluded that the studied population of blood donors is exposed to various risk factors associated with infection by T. gondii, and it is likely that the high frequency of anti-T. gondii IgG class found in this study is related to this exposure. The results show that there is no molecular evidence of parasitaemia in the studied donors, thus the risk of transmission of this infection via blood transfusion is low or zero in this region. In addition, the failure of genotyping and the non-occurrence of seroconversion of reanalyzed donors suggest that the positivity found in some samples analyzed by Nested PCR, were related to false-positive results.Introdução: Toxoplasma gondii, causador da toxoplasmose, é um protozoário intracelular que apresenta várias vias de transmissão, dentre elas a transfusional. Objetivo: Investigar por métodos sorológicos e moleculares, a infecção por T. gondii em doadores de sangue para avaliar o risco de transmissão via transfusional. Casuística e métodos: Foram selecionados 750 indivíduos aptos à doação de sangue. Estes indivíduos foram submetidos a uma entrevista sobre seus hábitos de vida e a uma coleta de sangue periférico. Com o ensaio imunoenzimático (ELISA) foi investigada a presença de anticorpos anti-T. gondii das classes IgM e IgG, e por Nested PCR e qPCR a parasitemia. As amostras positivas nos métodos moleculares foram submetidas à genotipagem pelo método Multilocus Nested PCR RFLP. Os doadores com resultados moleculares positivos e sorologias não reagentes foram reconvocados para investigar a soroconversão por ELISA e imunofluorescência indireta (IFI). Além destes, os receptores dos hemocomponentes com suspeita molecular de parasitemia foram rastreados. Destes, foram separadas alíquotas de sangue periférico, antes da transfusão e após a transfusão (±20 dias), as quais também foram analisadas pelos métodos sorológicos (ELISA e IFI) e moleculares (Nested PCR, qPCR e Mulitplex Nested PCR RFLP). Resultados: Trezentos e cinquenta e sete (47,6%) doadores apresentaram resultado reagente e 393 (52,4%) apresentaram resultado não reagente para a pesquisa de anticorpos anti-T. gondii da classe IgG. Vinte e sete (3,6%) apresentaram resultado reagente para IgM e 723 (96,4%) não reagente. As variáveis associadas a infecção por T. gondii foram: média de idade avançada (P < 0,0001), consumo de leite cru (P = 0,001), consumo de carne crua e mal passada de bovino e suíno (P = 0,003), residir na zona rural (P = 0,004), presença frequente de vetores mecânicos na residência (barata, rato e mosca P = 0,02; ratos, P = 0,03), menor grau de escolaridade (1º grau incompleto, P < 0,0001 e 1º grau completo, P = 0,002) e a baixa renda familiar (P = 0,002). Nenhuma amostra foi positiva pela qPCR, mas 40 (11%) apresentaram resultado positivo pela Nested PCR e sorologia reagente. Entretanto, estas amostras ao serem submetidas a genotipagem, não apresentaram amplificações. Os quatro casos rastreados também não apresentaram resultados positivos nas análises moleculares e nenhum doador reconvocado apresentou soroconversão. Conclusão: Concluímos que a população de doadores de sangue estudada está exposta a vários fatores de riscos associados a infecção por T. gondii e, é provável que a elevada frequência de anticorpos anti-T. gondii da classe IgG encontrada neste trabalho esteja relacionada a esta exposição. Os resultados demonstram que não há evidências moleculares de parasitemia nos doadores estudados, portanto o risco de transmissão desta infecção via transfusional é baixo ou nulo para esta região. Além disto, o insucesso da genotipagem e a não ocorrência de soroconversão dos doadores reanalisados sugerem que a positividade, encontrada em algumas amostras analisadas por Nested PCR, tratavam-se de resultados falso-positivos.Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2016-06-20T21:04:05Z No. of bitstreams: 1 fabiananakashima_tese.pdf: 1263029 bytes, checksum: ea1d05d8998fac5ccacd5e04039e5c2a (MD5)Made available in DSpace on 2016-06-20T21:04:05Z (GMT). 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dc.title.por.fl_str_mv Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
title Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
spellingShingle Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
Nakashima, Fabiana
Toxoplasma
Toxoplasmosis
Molecular Diagnostic Techniques
Serology
Toxoplasma
Toxoplasmose
Técnicas de Diagnóstico Molecular
Sorologia
CIENCIAS DA SAUDE::8765449414823306929::600
title_short Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
title_full Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
title_fullStr Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
title_full_unstemmed Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
title_sort Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue
author Nakashima, Fabiana
author_facet Nakashima, Fabiana
author_role author
dc.contributor.advisor1.fl_str_mv Mattos, Luiz Carlos de
dc.contributor.advisor-co1.fl_str_mv Mattos, Cinara de Cássia Brandão de
dc.contributor.referee1.fl_str_mv Castiglioni, Lilian
dc.contributor.referee2.fl_str_mv Ricci Junior, Octávio
dc.contributor.referee3.fl_str_mv Ferreira, Ana Iara da Costa
dc.contributor.referee4.fl_str_mv Martin, Natália
dc.contributor.authorID.fl_str_mv 34254202830
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9715943105309778
dc.contributor.author.fl_str_mv Nakashima, Fabiana
contributor_str_mv Mattos, Luiz Carlos de
Mattos, Cinara de Cássia Brandão de
Castiglioni, Lilian
Ricci Junior, Octávio
Ferreira, Ana Iara da Costa
Martin, Natália
dc.subject.eng.fl_str_mv Toxoplasma
Toxoplasmosis
Molecular Diagnostic Techniques
Serology
topic Toxoplasma
Toxoplasmosis
Molecular Diagnostic Techniques
Serology
Toxoplasma
Toxoplasmose
Técnicas de Diagnóstico Molecular
Sorologia
CIENCIAS DA SAUDE::8765449414823306929::600
dc.subject.por.fl_str_mv Toxoplasma
Toxoplasmose
Técnicas de Diagnóstico Molecular
Sorologia
dc.subject.cnpq.fl_str_mv CIENCIAS DA SAUDE::8765449414823306929::600
description Introduction: Toxoplasma gondii, which causes toxoplasmosis, is an intracellular protozoan that has several transmission routes, among them the transfusion. Objective: To investigate by serological and molecular methods the T. gondii infection in blood donors to assess the risk of transmission via transfusion. Methods: We selected 750 individuals able to donate blood. These individuals were submitted to an interview about their lifestyle habits and a collection of peripheral blood. The immunosorbent assay (ELISA) was used to investigate the presence of anti-T. gondii IgM and IgG classes, and the Nested PCR and qQPCR, the parasitaemia. Positive samples in the molecular methods were submitted for genotyping by the Multilocus Nested PCR RFLP method. Donors with positive molecular result and no positive serology were recalled to investigate seroconversion by ELISA and Indirect Immunofluorescence (IIF). In addition, recipients of blood components with molecular suspected parasitemia were screened. Of these, aliquots were separated from peripheral blood prior to transfusion and after transfusion (± 20 days), which were also analyzed by serological methods (ELISA and IIF) and molecular (PCR, qPCR Mulitplex nested PCR and RFLP). Results: Three hundred and fifty-seven (47.6%) donors had positive result and 393 (52.4%) showed no positive result for the presence of anti-T. gondii IgG class. Twenty-seven (3,6%) showed positive result for IgM and 723 (96.4%) non-reactive. The variables associated with infection by T. gondii were: advanced age (P < 0.0001), consumption of raw milk (P = 0.001), consumption of raw and underare beef and pork meat (P = 0.003), to reside in the countryside (P = 0.004), frequent presence of mechanical vectors in the residence (cockroach, mouse and fly P = 0.02; mice, P = 0.03), lower education (1st grade incomplete, P <0.0001 and 1st grade complete, P = 0.002) and low family income (P = 0.002). No sample was positive by qPCR, but 40 (11%) were positive by Nested PCR and positive serology. However, these samples did not show amplification when undergoing to genotyping. The four cases screened did not show positive results to molecular analysis and no recalled donor presented seroconversion. Conclusion: We concluded that the studied population of blood donors is exposed to various risk factors associated with infection by T. gondii, and it is likely that the high frequency of anti-T. gondii IgG class found in this study is related to this exposure. The results show that there is no molecular evidence of parasitaemia in the studied donors, thus the risk of transmission of this infection via blood transfusion is low or zero in this region. In addition, the failure of genotyping and the non-occurrence of seroconversion of reanalyzed donors suggest that the positivity found in some samples analyzed by Nested PCR, were related to false-positive results.
publishDate 2015
dc.date.issued.fl_str_mv 2015-12-14
dc.date.accessioned.fl_str_mv 2016-06-20T21:04:05Z
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dc.identifier.citation.fl_str_mv Nakashima, Fabiana. Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue. 2015. 118 p. Tese ( Programa de Pós-Graduação em Ciências da Saúde) - Faculdade de Medicina de São José do Rio Preto, São José do Rio Preto .
dc.identifier.uri.fl_str_mv http://bdtd.famerp.br/handle/tede/267
dc.identifier.doi.por.fl_str_mv 1159
identifier_str_mv Nakashima, Fabiana. Investigação sorológica e molecular de toxoplasma gondii em doadores de sangue. 2015. 118 p. Tese ( Programa de Pós-Graduação em Ciências da Saúde) - Faculdade de Medicina de São José do Rio Preto, São José do Rio Preto .
1159
url http://bdtd.famerp.br/handle/tede/267
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dc.publisher.department.fl_str_mv Faculdade 1::Departamento 1::306626487509624506::500
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