Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2022 |
| Outros Autores: | , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | por |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/51494 |
Resumo: | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Interação Parasito-Hospedeiro e Epidemiologia. Salvador, BA, Brasil. |
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Mota, Tiago FeitosaBrodskyn, Claudia IdaMorello, Luis GustavoMarchini, Fabricio KleryntonKrieger, Marco AurelioRampazzo, Rita de Cássia PontelloFraga, Deborah Bittencourt Mothé2022-02-25T15:16:00Z2022-02-25T15:16:00Z2022MOTA, Tiago Feitosa et al. Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples. Medical and Veterinary Entomology, p. 1-9, 2022.1365-2915https://www.arca.fiocruz.br/handle/icict/5149410.1111/mve.12564porWileymultiplex qPCRphlebotomineGenes InsectMultiplex Polymerase Chain ReactionLeishmaniasis, VisceralGenes de InsectoReacción en Cadena de la Polimerasa MultiplexLeishmaniasis VisceralGènes d'insecteRéaction de polymérisation en chaine multiplexLeishmaniose viscéraleGenes de InsetosReação em Cadeia da Polimerase MultiplexPhlebotominaeLeishmaniose VisceralMultiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samplesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Interação Parasito-Hospedeiro e Epidemiologia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Interação Parasito-Hospedeiro e Epidemiologia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil / Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil / Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Interação Parasito-Hospedeiro e Epidemiologia. Salvador, BA, Brasil / Universidade Federal da Bahia. Escola de Medicina Veterinária e Zootecnia. Departamento de Medicina Veterinária Preventiva e Produção Animal. Salvador, BA, Brasil.The study aimed to develop a multiplex qPCR to detect Leishmania infantum load in different sandfly sample settings using Leishmania kDNA and sandfly vacuolar ATPase (VATP) subunit C as internal control gene. The amplification of Lutzomyia longipalpis VATP gene was evaluated together with Leishmania infantum kDNA in a multiplex reaction. The concentration of VATP gene oligonucleotides was adjusted until no statistically significant difference was observed between all multiplex standard curves and singleplex curves, that is, only kDNA amplification. Limit of detection (LoD) was measured using a probit model and a cut-off defined by receiver operating characteristic analysis. Limit of quantification (LoQ) was assessed by a linear model using the coefficient of variation threshold of 25%. After assuring VATP gene amplification, its primer–probe concentrations were best at 100 nM/10 nM, respectively. The cut-off Cq value for L. infantum kDNA was defined as 35.46 with 100% of sensitivity and specificity. A total of 95% LoD was determined to be of 0.162 parasites while LoQ was 5.858. Our VATP/kDNA multiplex qPCR assay shows that it can be used to evaluate both DNA integrity and determine L. infantum load in L. longipalpis even for low yielded samples, that is, individual midguts.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/51494/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINALMedical Vet Entomology - 2022OK.pdfMedical Vet Entomology - 2022OK.pdfapplication/pdf984153https://www.arca.fiocruz.br/bitstream/icict/51494/2/Medical%20Vet%20Entomology%20-%202022OK.pdf1d015c00d967a57c7b7bdbad48728f50MD52TEXTMedical Vet Entomology - 2022OK.pdf.txtMedical Vet Entomology - 2022OK.pdf.txtExtracted texttext/plain43879https://www.arca.fiocruz.br/bitstream/icict/51494/3/Medical%20Vet%20Entomology%20-%202022OK.pdf.txt958382b7764affe49b7ebd390f607d64MD53icict/514942022-02-28 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| dc.title.pt_BR.fl_str_mv |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples |
| title |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples |
| spellingShingle |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples Mota, Tiago Feitosa multiplex qPCR phlebotomine Genes Insect Multiplex Polymerase Chain Reaction Leishmaniasis, Visceral Genes de Insecto Reacción en Cadena de la Polimerasa Multiplex Leishmaniasis Visceral Gènes d'insecte Réaction de polymérisation en chaine multiplex Leishmaniose viscérale Genes de Insetos Reação em Cadeia da Polimerase Multiplex Phlebotominae Leishmaniose Visceral |
| title_short |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples |
| title_full |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples |
| title_fullStr |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples |
| title_full_unstemmed |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples |
| title_sort |
Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples |
| author |
Mota, Tiago Feitosa |
| author_facet |
Mota, Tiago Feitosa Brodskyn, Claudia Ida Morello, Luis Gustavo Marchini, Fabricio Klerynton Krieger, Marco Aurelio Rampazzo, Rita de Cássia Pontello Fraga, Deborah Bittencourt Mothé |
| author_role |
author |
| author2 |
Brodskyn, Claudia Ida Morello, Luis Gustavo Marchini, Fabricio Klerynton Krieger, Marco Aurelio Rampazzo, Rita de Cássia Pontello Fraga, Deborah Bittencourt Mothé |
| author2_role |
author author author author author author |
| dc.contributor.author.fl_str_mv |
Mota, Tiago Feitosa Brodskyn, Claudia Ida Morello, Luis Gustavo Marchini, Fabricio Klerynton Krieger, Marco Aurelio Rampazzo, Rita de Cássia Pontello Fraga, Deborah Bittencourt Mothé |
| dc.subject.other.pt_BR.fl_str_mv |
multiplex qPCR phlebotomine |
| topic |
multiplex qPCR phlebotomine Genes Insect Multiplex Polymerase Chain Reaction Leishmaniasis, Visceral Genes de Insecto Reacción en Cadena de la Polimerasa Multiplex Leishmaniasis Visceral Gènes d'insecte Réaction de polymérisation en chaine multiplex Leishmaniose viscérale Genes de Insetos Reação em Cadeia da Polimerase Multiplex Phlebotominae Leishmaniose Visceral |
| dc.subject.en.pt_BR.fl_str_mv |
Genes Insect Multiplex Polymerase Chain Reaction Leishmaniasis, Visceral |
| dc.subject.es.pt_BR.fl_str_mv |
Genes de Insecto Reacción en Cadena de la Polimerasa Multiplex Leishmaniasis Visceral |
| dc.subject.fr.pt_BR.fl_str_mv |
Gènes d'insecte Réaction de polymérisation en chaine multiplex Leishmaniose viscérale |
| dc.subject.decs.pt_BR.fl_str_mv |
Genes de Insetos Reação em Cadeia da Polimerase Multiplex Phlebotominae Leishmaniose Visceral |
| description |
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Interação Parasito-Hospedeiro e Epidemiologia. Salvador, BA, Brasil. |
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2022 |
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2022-02-25T15:16:00Z |
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2022-02-25T15:16:00Z |
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2022 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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MOTA, Tiago Feitosa et al. Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples. Medical and Veterinary Entomology, p. 1-9, 2022. |
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https://www.arca.fiocruz.br/handle/icict/51494 |
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1365-2915 |
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10.1111/mve.12564 |
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MOTA, Tiago Feitosa et al. Multiplex qPCR assay to determine Leishmania infantum load in Lutzomyia longipalpis sandfly samples. Medical and Veterinary Entomology, p. 1-9, 2022. 1365-2915 10.1111/mve.12564 |
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Wiley |
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