Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection
Autor(a) principal: | |
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Data de Publicação: | 2010 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Digital do Instituto Evandro Chagas (Patuá) |
Texto Completo: | https://patua.iec.gov.br/handle/iec/733 |
Resumo: | There is a need for standardization and simplification of the existing methods for molecular detection of Leishmania infantum in the canine reservoir host. The commercially available OligoC-TesT kit incorporates standardized PCR reagents with rapid oligochromatographic dipstick detection of PCR products and is highly sensitive for use in humans but not yet independently validated for use in dogs. Here we compare the sensitivity of OligoC-TesT with those of nested kinetoplast DNA (kDNA) PCR, nested internal transcribed spacer 1 (ITS-1) PCR, and a PCR-hybridization protocol, using longitudinal naturally infected canine bone marrow samples whose parasite burdens were measured by real-time quantitative PCR (qPCR). The sensitivity of OligoC-TesT for infected dogs was 70% (95% confidence interval [CI], 63 to 78%), similar to that of kDNA PCR (72%; 95% CI, 65 to 80%; P 0.69) but significantly greater than those of PCR-hybridization (61%; 95% CI, 53 to 69%; P 0.007) and ITS-1 nested PCR (54%; 95% CI, 45 to 62%; P < 0.001); real-time qPCR had the highest sensitivity (91%; 95% CI, 85 to 95%; P < 0.001). OligoC-TesT sensitivity was greater for polysymptomatic and oligosymptomatic dogs than for asymptomatic dogs (93%, 74%, and 61%, respectively; P 0.005), a trend also observed for the other qualitative PCR methods tested (P < 0.05). Test positivity increased with increasing parasite burdens, as measured by real-time qPCR: OligoC-TesT and kDNA PCR detected 100% and 99% of positive samples when parasite burdens exceeded 74 and 49 parasites/ml, respectively. OligoC-TesT has high sensitivity for detection of canine Leishmania infections; its ease of operation and ease of interpretation are further advantages for veterinary diagnostic laboratories and for large-scale survey work in developing countries. |
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Carson, CannorQuinnell, Rupert JHolden, JenniferSantos, Lourdes Maria Garcez dosDeborggraeve, StijinCourtenay, Orin2016-01-26T11:36:46Z2016-01-26T11:36:46Z2010CARSON, Cannor et al. Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection. Journal of Clinical Microbiology, v. 48, n. 9, p. 3325-3350, Sept. 20101098-660Xhttps://patua.iec.gov.br/handle/iec/73310.1128/JCM.02331-09There is a need for standardization and simplification of the existing methods for molecular detection of Leishmania infantum in the canine reservoir host. The commercially available OligoC-TesT kit incorporates standardized PCR reagents with rapid oligochromatographic dipstick detection of PCR products and is highly sensitive for use in humans but not yet independently validated for use in dogs. Here we compare the sensitivity of OligoC-TesT with those of nested kinetoplast DNA (kDNA) PCR, nested internal transcribed spacer 1 (ITS-1) PCR, and a PCR-hybridization protocol, using longitudinal naturally infected canine bone marrow samples whose parasite burdens were measured by real-time quantitative PCR (qPCR). The sensitivity of OligoC-TesT for infected dogs was 70% (95% confidence interval [CI], 63 to 78%), similar to that of kDNA PCR (72%; 95% CI, 65 to 80%; P 0.69) but significantly greater than those of PCR-hybridization (61%; 95% CI, 53 to 69%; P 0.007) and ITS-1 nested PCR (54%; 95% CI, 45 to 62%; P < 0.001); real-time qPCR had the highest sensitivity (91%; 95% CI, 85 to 95%; P < 0.001). OligoC-TesT sensitivity was greater for polysymptomatic and oligosymptomatic dogs than for asymptomatic dogs (93%, 74%, and 61%, respectively; P 0.005), a trend also observed for the other qualitative PCR methods tested (P < 0.05). Test positivity increased with increasing parasite burdens, as measured by real-time qPCR: OligoC-TesT and kDNA PCR detected 100% and 99% of positive samples when parasite burdens exceeded 74 and 49 parasites/ml, respectively. OligoC-TesT has high sensitivity for detection of canine Leishmania infections; its ease of operation and ease of interpretation are further advantages for veterinary diagnostic laboratories and for large-scale survey work in developing countries.University of Warwick. Department of Biological Sciences. Coventry, United Kingdom.University of Leeds. Institute of Integrative and Comparative Biology. Leeds, United Kingdom.University of Warwick. Department of Biological Sciences. Coventry, United Kingdom.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Belém, PA, Brasil.Institute of Tropical Medicine. Antwerp, Belgium.University of Warwick. Department of Biological Sciences. 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dc.title.pt_BR.fl_str_mv |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection |
title |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection |
spellingShingle |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection Carson, Cannor Leishmania infantum / genética Leishmania infantum / isolamento & purificação Hibridização de Ácido Nucleico / métodos Doenças do Cão / diagnóstico Cães Parasitologia / métodos Sensibilidade e Especificidade |
title_short |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection |
title_full |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection |
title_fullStr |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection |
title_full_unstemmed |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection |
title_sort |
Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection |
author |
Carson, Cannor |
author_facet |
Carson, Cannor Quinnell, Rupert J Holden, Jennifer Santos, Lourdes Maria Garcez dos Deborggraeve, Stijin Courtenay, Orin |
author_role |
author |
author2 |
Quinnell, Rupert J Holden, Jennifer Santos, Lourdes Maria Garcez dos Deborggraeve, Stijin Courtenay, Orin |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Carson, Cannor Quinnell, Rupert J Holden, Jennifer Santos, Lourdes Maria Garcez dos Deborggraeve, Stijin Courtenay, Orin |
dc.subject.decsPrimary.pt_BR.fl_str_mv |
Leishmania infantum / genética Leishmania infantum / isolamento & purificação Hibridização de Ácido Nucleico / métodos Doenças do Cão / diagnóstico Cães Parasitologia / métodos Sensibilidade e Especificidade |
topic |
Leishmania infantum / genética Leishmania infantum / isolamento & purificação Hibridização de Ácido Nucleico / métodos Doenças do Cão / diagnóstico Cães Parasitologia / métodos Sensibilidade e Especificidade |
description |
There is a need for standardization and simplification of the existing methods for molecular detection of Leishmania infantum in the canine reservoir host. The commercially available OligoC-TesT kit incorporates standardized PCR reagents with rapid oligochromatographic dipstick detection of PCR products and is highly sensitive for use in humans but not yet independently validated for use in dogs. Here we compare the sensitivity of OligoC-TesT with those of nested kinetoplast DNA (kDNA) PCR, nested internal transcribed spacer 1 (ITS-1) PCR, and a PCR-hybridization protocol, using longitudinal naturally infected canine bone marrow samples whose parasite burdens were measured by real-time quantitative PCR (qPCR). The sensitivity of OligoC-TesT for infected dogs was 70% (95% confidence interval [CI], 63 to 78%), similar to that of kDNA PCR (72%; 95% CI, 65 to 80%; P 0.69) but significantly greater than those of PCR-hybridization (61%; 95% CI, 53 to 69%; P 0.007) and ITS-1 nested PCR (54%; 95% CI, 45 to 62%; P < 0.001); real-time qPCR had the highest sensitivity (91%; 95% CI, 85 to 95%; P < 0.001). OligoC-TesT sensitivity was greater for polysymptomatic and oligosymptomatic dogs than for asymptomatic dogs (93%, 74%, and 61%, respectively; P 0.005), a trend also observed for the other qualitative PCR methods tested (P < 0.05). Test positivity increased with increasing parasite burdens, as measured by real-time qPCR: OligoC-TesT and kDNA PCR detected 100% and 99% of positive samples when parasite burdens exceeded 74 and 49 parasites/ml, respectively. OligoC-TesT has high sensitivity for detection of canine Leishmania infections; its ease of operation and ease of interpretation are further advantages for veterinary diagnostic laboratories and for large-scale survey work in developing countries. |
publishDate |
2010 |
dc.date.issued.fl_str_mv |
2010 |
dc.date.accessioned.fl_str_mv |
2016-01-26T11:36:46Z |
dc.date.available.fl_str_mv |
2016-01-26T11:36:46Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
CARSON, Cannor et al. Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection. Journal of Clinical Microbiology, v. 48, n. 9, p. 3325-3350, Sept. 2010 |
dc.identifier.uri.fl_str_mv |
https://patua.iec.gov.br/handle/iec/733 |
dc.identifier.issn.-.fl_str_mv |
1098-660X |
dc.identifier.doi.-.fl_str_mv |
10.1128/JCM.02331-09 |
identifier_str_mv |
CARSON, Cannor et al. Comparison of Leishmania OligoC-TesT PCR with conventional and real-time PCR for diagnosis of canine leishmania infection. Journal of Clinical Microbiology, v. 48, n. 9, p. 3325-3350, Sept. 2010 1098-660X 10.1128/JCM.02331-09 |
url |
https://patua.iec.gov.br/handle/iec/733 |
dc.language.iso.fl_str_mv |
eng |
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eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
American Society for Microbiology |
publisher.none.fl_str_mv |
American Society for Microbiology |
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