Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR
Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classific...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Wouter Graumans [verfasserIn] Fitsum G. Tadesse [verfasserIn] Chiara Andolina [verfasserIn] Geert-Jan van Gemert [verfasserIn] Karina Teelen [verfasserIn] Kjerstin Lanke [verfasserIn] Endalamaw Gadisa [verfasserIn] Delenasaw Yewhalaw [verfasserIn] Marga van de Vegte-Bolmer [verfasserIn] Rianne Siebelink-Stoter [verfasserIn] Isaïe Reuling [verfasserIn] Robert Sauerwein [verfasserIn] Teun Bousema [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2017 |
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| Schlagwörter: |
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| Übergeordnetes Werk: |
In: Malaria Journal - BMC, 2003, 16(2017), 1, Seite 12 |
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| Übergeordnetes Werk: |
volume:16 ; year:2017 ; number:1 ; pages:12 |
| Links: |
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| DOI / URN: |
10.1186/s12936-017-2011-9 |
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| Katalog-ID: |
DOAJ02897557X |
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| 245 | 1 | 0 | |a Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR |
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| 520 | |a Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. | ||
| 650 | 4 | |a Transmission | |
| 650 | 4 | |a Oocyst | |
| 650 | 4 | |a Sporozoite | |
| 650 | 4 | |a Anopheles | |
| 650 | 4 | |a Infectivity | |
| 650 | 4 | |a Gametocyte | |
| 653 | 0 | |a Arctic medicine. Tropical medicine | |
| 653 | 0 | |a Infectious and parasitic diseases | |
| 700 | 0 | |a Fitsum G. Tadesse |e verfasserin |4 aut | |
| 700 | 0 | |a Chiara Andolina |e verfasserin |4 aut | |
| 700 | 0 | |a Geert-Jan van Gemert |e verfasserin |4 aut | |
| 700 | 0 | |a Karina Teelen |e verfasserin |4 aut | |
| 700 | 0 | |a Kjerstin Lanke |e verfasserin |4 aut | |
| 700 | 0 | |a Endalamaw Gadisa |e verfasserin |4 aut | |
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| 700 | 0 | |a Marga van de Vegte-Bolmer |e verfasserin |4 aut | |
| 700 | 0 | |a Rianne Siebelink-Stoter |e verfasserin |4 aut | |
| 700 | 0 | |a Isaïe Reuling |e verfasserin |4 aut | |
| 700 | 0 | |a Robert Sauerwein |e verfasserin |4 aut | |
| 700 | 0 | |a Teun Bousema |e verfasserin |4 aut | |
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10.1186/s12936-017-2011-9 doi (DE-627)DOAJ02897557X (DE-599)DOAJ984417d7182e49a2a44fc7dcbcbc8b06 DE-627 ger DE-627 rakwb eng RC955-962 RC109-216 Wouter Graumans verfasserin aut Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. Transmission Oocyst Sporozoite Anopheles Infectivity Gametocyte Arctic medicine. Tropical medicine Infectious and parasitic diseases Fitsum G. Tadesse verfasserin aut Chiara Andolina verfasserin aut Geert-Jan van Gemert verfasserin aut Karina Teelen verfasserin aut Kjerstin Lanke verfasserin aut Endalamaw Gadisa verfasserin aut Delenasaw Yewhalaw verfasserin aut Marga van de Vegte-Bolmer verfasserin aut Rianne Siebelink-Stoter verfasserin aut Isaïe Reuling verfasserin aut Robert Sauerwein verfasserin aut Teun Bousema verfasserin aut In Malaria Journal BMC, 2003 16(2017), 1, Seite 12 (DE-627)355986582 (DE-600)2091229-8 14752875 nnns volume:16 year:2017 number:1 pages:12 https://doi.org/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/article/984417d7182e49a2a44fc7dcbcbc8b06 kostenfrei http://link.springer.com/article/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/toc/1475-2875 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 16 2017 1 12 |
| spelling |
10.1186/s12936-017-2011-9 doi (DE-627)DOAJ02897557X (DE-599)DOAJ984417d7182e49a2a44fc7dcbcbc8b06 DE-627 ger DE-627 rakwb eng RC955-962 RC109-216 Wouter Graumans verfasserin aut Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. Transmission Oocyst Sporozoite Anopheles Infectivity Gametocyte Arctic medicine. Tropical medicine Infectious and parasitic diseases Fitsum G. Tadesse verfasserin aut Chiara Andolina verfasserin aut Geert-Jan van Gemert verfasserin aut Karina Teelen verfasserin aut Kjerstin Lanke verfasserin aut Endalamaw Gadisa verfasserin aut Delenasaw Yewhalaw verfasserin aut Marga van de Vegte-Bolmer verfasserin aut Rianne Siebelink-Stoter verfasserin aut Isaïe Reuling verfasserin aut Robert Sauerwein verfasserin aut Teun Bousema verfasserin aut In Malaria Journal BMC, 2003 16(2017), 1, Seite 12 (DE-627)355986582 (DE-600)2091229-8 14752875 nnns volume:16 year:2017 number:1 pages:12 https://doi.org/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/article/984417d7182e49a2a44fc7dcbcbc8b06 kostenfrei http://link.springer.com/article/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/toc/1475-2875 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 16 2017 1 12 |
| allfields_unstemmed |
10.1186/s12936-017-2011-9 doi (DE-627)DOAJ02897557X (DE-599)DOAJ984417d7182e49a2a44fc7dcbcbc8b06 DE-627 ger DE-627 rakwb eng RC955-962 RC109-216 Wouter Graumans verfasserin aut Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. Transmission Oocyst Sporozoite Anopheles Infectivity Gametocyte Arctic medicine. Tropical medicine Infectious and parasitic diseases Fitsum G. Tadesse verfasserin aut Chiara Andolina verfasserin aut Geert-Jan van Gemert verfasserin aut Karina Teelen verfasserin aut Kjerstin Lanke verfasserin aut Endalamaw Gadisa verfasserin aut Delenasaw Yewhalaw verfasserin aut Marga van de Vegte-Bolmer verfasserin aut Rianne Siebelink-Stoter verfasserin aut Isaïe Reuling verfasserin aut Robert Sauerwein verfasserin aut Teun Bousema verfasserin aut In Malaria Journal BMC, 2003 16(2017), 1, Seite 12 (DE-627)355986582 (DE-600)2091229-8 14752875 nnns volume:16 year:2017 number:1 pages:12 https://doi.org/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/article/984417d7182e49a2a44fc7dcbcbc8b06 kostenfrei http://link.springer.com/article/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/toc/1475-2875 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 16 2017 1 12 |
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10.1186/s12936-017-2011-9 doi (DE-627)DOAJ02897557X (DE-599)DOAJ984417d7182e49a2a44fc7dcbcbc8b06 DE-627 ger DE-627 rakwb eng RC955-962 RC109-216 Wouter Graumans verfasserin aut Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. Transmission Oocyst Sporozoite Anopheles Infectivity Gametocyte Arctic medicine. Tropical medicine Infectious and parasitic diseases Fitsum G. Tadesse verfasserin aut Chiara Andolina verfasserin aut Geert-Jan van Gemert verfasserin aut Karina Teelen verfasserin aut Kjerstin Lanke verfasserin aut Endalamaw Gadisa verfasserin aut Delenasaw Yewhalaw verfasserin aut Marga van de Vegte-Bolmer verfasserin aut Rianne Siebelink-Stoter verfasserin aut Isaïe Reuling verfasserin aut Robert Sauerwein verfasserin aut Teun Bousema verfasserin aut In Malaria Journal BMC, 2003 16(2017), 1, Seite 12 (DE-627)355986582 (DE-600)2091229-8 14752875 nnns volume:16 year:2017 number:1 pages:12 https://doi.org/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/article/984417d7182e49a2a44fc7dcbcbc8b06 kostenfrei http://link.springer.com/article/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/toc/1475-2875 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 16 2017 1 12 |
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10.1186/s12936-017-2011-9 doi (DE-627)DOAJ02897557X (DE-599)DOAJ984417d7182e49a2a44fc7dcbcbc8b06 DE-627 ger DE-627 rakwb eng RC955-962 RC109-216 Wouter Graumans verfasserin aut Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. Transmission Oocyst Sporozoite Anopheles Infectivity Gametocyte Arctic medicine. Tropical medicine Infectious and parasitic diseases Fitsum G. Tadesse verfasserin aut Chiara Andolina verfasserin aut Geert-Jan van Gemert verfasserin aut Karina Teelen verfasserin aut Kjerstin Lanke verfasserin aut Endalamaw Gadisa verfasserin aut Delenasaw Yewhalaw verfasserin aut Marga van de Vegte-Bolmer verfasserin aut Rianne Siebelink-Stoter verfasserin aut Isaïe Reuling verfasserin aut Robert Sauerwein verfasserin aut Teun Bousema verfasserin aut In Malaria Journal BMC, 2003 16(2017), 1, Seite 12 (DE-627)355986582 (DE-600)2091229-8 14752875 nnns volume:16 year:2017 number:1 pages:12 https://doi.org/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/article/984417d7182e49a2a44fc7dcbcbc8b06 kostenfrei http://link.springer.com/article/10.1186/s12936-017-2011-9 kostenfrei https://doaj.org/toc/1475-2875 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 16 2017 1 12 |
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Wouter Graumans @@aut@@ Fitsum G. Tadesse @@aut@@ Chiara Andolina @@aut@@ Geert-Jan van Gemert @@aut@@ Karina Teelen @@aut@@ Kjerstin Lanke @@aut@@ Endalamaw Gadisa @@aut@@ Delenasaw Yewhalaw @@aut@@ Marga van de Vegte-Bolmer @@aut@@ Rianne Siebelink-Stoter @@aut@@ Isaïe Reuling @@aut@@ Robert Sauerwein @@aut@@ Teun Bousema @@aut@@ |
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2017-01-01T00:00:00Z |
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RC955-962 RC109-216 Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR Transmission Oocyst Sporozoite Anopheles Infectivity Gametocyte |
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Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR |
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Wouter Graumans Fitsum G. Tadesse Chiara Andolina Geert-Jan van Gemert Karina Teelen Kjerstin Lanke Endalamaw Gadisa Delenasaw Yewhalaw Marga van de Vegte-Bolmer Rianne Siebelink-Stoter Isaïe Reuling Robert Sauerwein Teun Bousema |
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semi-high-throughput detection of plasmodium falciparum and plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite elisa and quantitative pcr |
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Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR |
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Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. |
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Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. |
| abstract_unstemmed |
Abstract Background The malaria infection status of mosquitoes is commonly determined by microscopic detection of oocysts on the dissected mosquito midgut. This method is labour-intensive, does not allow processing of large numbers of mosquitoes and can be challenging in terms of objective classification of oocysts. Here, a semi-high-throughput bead-beating ELISA method is proposed for detection of the circumsporozoite protein (CSP) followed by confirmation by quantitative PCR (qPCR). Methods Cultured Plasmodium falciparum gametocytes were offered to Anopheles stephensi mosquitoes and examined by microscopy. After bead-beating, mosquito homogenate was examined by CSP-ELISA and 18S qPCR. As negative controls, mosquitoes that were offered a heat-inactivated gametocyte blood meal were used. The CSP-ELISA/qPCR methodology was applied to high and low-intensity infections of cultured P. falciparum gametocytes. A similar methodology optimized for P. vivax was used on mosquitoes that were offered blood from Ethiopian donors who were naturally infected with P. vivax. Results There was considerable variation in CSP-ELISA signal and qPCR values in mosquitoes with low oocyst intensities. There was a strong agreement mosquito positivity by CSP-ELISA and by qPCR in mosquitoes that fed on cultured P. falciparum material (agreement 96.9%; kappa = 0.97) and naturally infected P. vivax parasite carriers [agreement 92.4% (kappa = 0.83)]. Conclusions The proposed bead-beating CSP-ELISA/qPCR methodology considerably increases throughput for the detection of mosquito infection. qPCR remains necessary to confirm infections in mosquitoes with low CSP-ELISA signal. This methodology may prove particularly useful for studies where very low mosquito infection prevalence is expected and study sites where experience with oocyst detection is limited. |
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Semi-high-throughput detection of Plasmodium falciparum and Plasmodium vivax oocysts in mosquitoes using bead-beating followed by circumsporozoite ELISA and quantitative PCR |
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Fitsum G. Tadesse Chiara Andolina Geert-Jan van Gemert Karina Teelen Kjerstin Lanke Endalamaw Gadisa Delenasaw Yewhalaw Marga van de Vegte-Bolmer Rianne Siebelink-Stoter Isaïe Reuling Robert Sauerwein Teun Bousema |
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