A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spots
Abstract Background Mansonella ozzardi is a poorly understood human filarial parasite with a broad distribution throughout Latin America. Most of what is known about its parasitism has come from epidemiological studies that have estimated parasite incidence using light microscopy. Light microscopy c...
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| Format: | Article |
| Language: | English |
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BMC
2015-05-01
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| Series: | Parasites & Vectors |
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| Online Access: | https://doi.org/10.1186/s13071-015-0889-z |
| _version_ | 1797811854630715392 |
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| author | Jansen Fernandes Medeiros Tatiana Amaral Pires Almeida Lucyane Bastos Tavares Silva Jose Miguel Rubio James Lee Crainey Felipe Arley Costa Pessoa Sergio Luiz Bessa Luz |
| author_facet | Jansen Fernandes Medeiros Tatiana Amaral Pires Almeida Lucyane Bastos Tavares Silva Jose Miguel Rubio James Lee Crainey Felipe Arley Costa Pessoa Sergio Luiz Bessa Luz |
| author_sort | Jansen Fernandes Medeiros |
| collection | DOAJ |
| description | Abstract Background Mansonella ozzardi is a poorly understood human filarial parasite with a broad distribution throughout Latin America. Most of what is known about its parasitism has come from epidemiological studies that have estimated parasite incidence using light microscopy. Light microscopy can, however, miss lighter, submicroscopic, infections. In this study we have compared M. ozzardi incidence estimates made using light microscopy, with estimates made using PCR. Methods 214 DNA extracts made from Large Volume Venous Blood Samples (LVVBS) were taken from volunteers from two study sites in the Rio Solimões region: Codajás [n = 109] and Tefé [n = 105] and were subsequently assayed for M. ozzardi parasitism using a diagnostic PCR (Mo-dPCR). Peripheral finger-prick blood samples were taken from the same individuals and used for microscopic examination. Finger-prick blood, taken from individuals from Tefé, was also used for the creation of FTA®card dried blood spots (DBS) that were subsequently subjected to Mo-dPCR. Results Overall M. ozzardi incidence estimates made with LVVBS PCRs were 1.8 times higher than those made using microscopy (44.9 % [96/214] compared with 24.3 % [52/214]) and 1.5 times higher than the PCR estimates made from FTA®card DBS (48/105 versus 31/105). PCR-based detection of FTA®card DBS proved 1.3 times more sensitive at diagnosing infections from peripheral blood samples than light microscopy did: detecting 24/105 compared with 31/105. PCR of LVVBS reported the fewest number of false negatives, detecting: 44 of 52 (84.6 %) individuals diagnosed by microscopy; 27 of 31 (87.1 %) of those diagnosed positive from DBSs and 17 out of 18 (94.4 %) of those diagnosed as positive by both alternative methodologies. Conclusions In this study, Mo-dPCR of LVVBS was by far the most sensitive method of detecting M. ozzardi infections and detected submicroscopic infections. Mo-dPCR FTA®card DBS also provided a more sensitive test for M. ozzardi diagnosis than light microscopy based diagnosis did and thus in settings where only finger-prick assays can be carried-out, it may be a more reliable method of detection. Most existing M. ozzardi incidence estimates, which are often based on light microscope diagnosis, are likely to dramatically underestimate true M. ozzardi parasitism incidence levels. |
| first_indexed | 2024-03-13T07:29:55Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-03-13T07:29:55Z |
| publishDate | 2015-05-01 |
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| series | Parasites & Vectors |
| spelling | doaj.art-dfa91cba7bc6410fa7f620843ae5b5c02023-06-04T11:12:01ZengBMCParasites & Vectors1756-33052015-05-01811810.1186/s13071-015-0889-zA field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spotsJansen Fernandes Medeiros0Tatiana Amaral Pires Almeida1Lucyane Bastos Tavares Silva2Jose Miguel Rubio3James Lee Crainey4Felipe Arley Costa Pessoa5Sergio Luiz Bessa Luz6Laboratory of Entomology, Fiocruz RondôniaResearch Programme on Infectious Disease Ecology in the Amazon (RP-IDEA), Leônidas & Maria Deane Institute – Fiocruz AmazôniaResearch Programme on Infectious Disease Ecology in the Amazon (RP-IDEA), Leônidas & Maria Deane Institute – Fiocruz AmazôniaMalaria & Emerging Parasitic Diseases Laboratory, Parasitology Department, National Centre of Microbiology. Carlos III Health InstituteResearch Programme on Infectious Disease Ecology in the Amazon (RP-IDEA), Leônidas & Maria Deane Institute – Fiocruz AmazôniaResearch Programme on Infectious Disease Ecology in the Amazon (RP-IDEA), Leônidas & Maria Deane Institute – Fiocruz AmazôniaResearch Programme on Infectious Disease Ecology in the Amazon (RP-IDEA), Leônidas & Maria Deane Institute – Fiocruz AmazôniaAbstract Background Mansonella ozzardi is a poorly understood human filarial parasite with a broad distribution throughout Latin America. Most of what is known about its parasitism has come from epidemiological studies that have estimated parasite incidence using light microscopy. Light microscopy can, however, miss lighter, submicroscopic, infections. In this study we have compared M. ozzardi incidence estimates made using light microscopy, with estimates made using PCR. Methods 214 DNA extracts made from Large Volume Venous Blood Samples (LVVBS) were taken from volunteers from two study sites in the Rio Solimões region: Codajás [n = 109] and Tefé [n = 105] and were subsequently assayed for M. ozzardi parasitism using a diagnostic PCR (Mo-dPCR). Peripheral finger-prick blood samples were taken from the same individuals and used for microscopic examination. Finger-prick blood, taken from individuals from Tefé, was also used for the creation of FTA®card dried blood spots (DBS) that were subsequently subjected to Mo-dPCR. Results Overall M. ozzardi incidence estimates made with LVVBS PCRs were 1.8 times higher than those made using microscopy (44.9 % [96/214] compared with 24.3 % [52/214]) and 1.5 times higher than the PCR estimates made from FTA®card DBS (48/105 versus 31/105). PCR-based detection of FTA®card DBS proved 1.3 times more sensitive at diagnosing infections from peripheral blood samples than light microscopy did: detecting 24/105 compared with 31/105. PCR of LVVBS reported the fewest number of false negatives, detecting: 44 of 52 (84.6 %) individuals diagnosed by microscopy; 27 of 31 (87.1 %) of those diagnosed positive from DBSs and 17 out of 18 (94.4 %) of those diagnosed as positive by both alternative methodologies. Conclusions In this study, Mo-dPCR of LVVBS was by far the most sensitive method of detecting M. ozzardi infections and detected submicroscopic infections. Mo-dPCR FTA®card DBS also provided a more sensitive test for M. ozzardi diagnosis than light microscopy based diagnosis did and thus in settings where only finger-prick assays can be carried-out, it may be a more reliable method of detection. Most existing M. ozzardi incidence estimates, which are often based on light microscope diagnosis, are likely to dramatically underestimate true M. ozzardi parasitism incidence levels.https://doi.org/10.1186/s13071-015-0889-zMansonelliasisMansonella ozzardiPCR-detectionSubmicroscopicFTA®cards |
| spellingShingle | Jansen Fernandes Medeiros Tatiana Amaral Pires Almeida Lucyane Bastos Tavares Silva Jose Miguel Rubio James Lee Crainey Felipe Arley Costa Pessoa Sergio Luiz Bessa Luz A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spots Parasites & Vectors Mansonelliasis Mansonella ozzardi PCR-detection Submicroscopic FTA®cards |
| title | A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spots |
| title_full | A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spots |
| title_fullStr | A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spots |
| title_full_unstemmed | A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spots |
| title_short | A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA® card dried blood spots |
| title_sort | field trial of a pcr based mansonella ozzardi diagnosis assay detects high levels of submicroscopic m ozzardi infections in both venous blood samples and fta r card dried blood spots |
| topic | Mansonelliasis Mansonella ozzardi PCR-detection Submicroscopic FTA®cards |
| url | https://doi.org/10.1186/s13071-015-0889-z |
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