Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR
Abstract Background Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription...
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ftdoajarticles:oai:doaj.org/article:587755320969404ab3e7ca1ef1a93d44 2023-05-15T15:18:05+02:00 Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR Kuamsab Napaporn Putaporntip Chaturong Pattanawong Urassaya Jongwutiwes Somchai 2012-06-01T00:00:00Z https://doi.org/10.1186/1475-2875-11-190 https://doaj.org/article/587755320969404ab3e7ca1ef1a93d44 EN eng BMC http://www.malariajournal.com/content/11/1/190 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-190 1475-2875 https://doaj.org/article/587755320969404ab3e7ca1ef1a93d44 Malaria Journal, Vol 11, Iss 1, p 190 (2012) Malaria diagnosis Gametocyte Reverse transcription polymerase chain reaction Plasmodium falciparum Plasmodium vivax Pfs25 Pvs25 Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2012 ftdoajarticles https://doi.org/10.1186/1475-2875-11-190 2022-12-31T08:53:16Z Abstract Background Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription polymerase chain reaction (RT-PCR) targeting gametocyte-specific mRNA. To date, RT-PCR has mainly been applied to the diagnosis of Plasmodium falciparum gametocytes but very limited for that of Plasmodium vivax . Methods A multiplex-nested RT-PCR targeting Pfs25 and Pvs25 mRNA specific to mature gametocytes of P. falciparum and P. vivax , respectively, was developed. The assay was evaluated using blood samples collected in rainy and dry seasons from febrile patients,in a malaria-endemic area in Thailand. Malaria diagnosis was performed by Giemsa-stained blood smears and 18S rRNA PCR. Results The multiplex-nested RT-PCR detected Pfs25 mRNA in 75 of 86 (87.2%) P. falciparum -infected individuals and Pvs25 mRNA in 82 of 90 (91.1%) P. vivax malaria patients diagnosed by 18S rRNA PCR. Gametocytes were detected in 38 (eight P. falciparum and 30 P. vivax ) of 157 microscopy positive samples, implying that a large number of patients harbour sub-microscopic gametocytaemia. No seasonal differences in gametocyte carriage were observed for both malaria species diagnosed by multiplex-nested RT-PCR. With single-nested RT-PCR targeting Pfs25 or Pvs25 mRNA as standard, the multiplex-nested RT-PCR offered sensitivities of 97.4% and 98.9% and specificities of 100% and 98.8% for diagnosing mature gametocytes of P. falciparum and P. vivax, respectively. The minimum detection limit of the multiplex-nested PCR was 10 copies of templates. Conclusions The multiplex-nested RT-PCR developed herein is useful for simultaneous assessment of both P. falciparum and P. vivax gametocyte carriage that is prevalent and generally sympatric in several malaria-endemic areas outside Africa. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 11 1 190 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
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Malaria diagnosis Gametocyte Reverse transcription polymerase chain reaction Plasmodium falciparum Plasmodium vivax Pfs25 Pvs25 Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Malaria diagnosis Gametocyte Reverse transcription polymerase chain reaction Plasmodium falciparum Plasmodium vivax Pfs25 Pvs25 Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Kuamsab Napaporn Putaporntip Chaturong Pattanawong Urassaya Jongwutiwes Somchai Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR |
topic_facet |
Malaria diagnosis Gametocyte Reverse transcription polymerase chain reaction Plasmodium falciparum Plasmodium vivax Pfs25 Pvs25 Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription polymerase chain reaction (RT-PCR) targeting gametocyte-specific mRNA. To date, RT-PCR has mainly been applied to the diagnosis of Plasmodium falciparum gametocytes but very limited for that of Plasmodium vivax . Methods A multiplex-nested RT-PCR targeting Pfs25 and Pvs25 mRNA specific to mature gametocytes of P. falciparum and P. vivax , respectively, was developed. The assay was evaluated using blood samples collected in rainy and dry seasons from febrile patients,in a malaria-endemic area in Thailand. Malaria diagnosis was performed by Giemsa-stained blood smears and 18S rRNA PCR. Results The multiplex-nested RT-PCR detected Pfs25 mRNA in 75 of 86 (87.2%) P. falciparum -infected individuals and Pvs25 mRNA in 82 of 90 (91.1%) P. vivax malaria patients diagnosed by 18S rRNA PCR. Gametocytes were detected in 38 (eight P. falciparum and 30 P. vivax ) of 157 microscopy positive samples, implying that a large number of patients harbour sub-microscopic gametocytaemia. No seasonal differences in gametocyte carriage were observed for both malaria species diagnosed by multiplex-nested RT-PCR. With single-nested RT-PCR targeting Pfs25 or Pvs25 mRNA as standard, the multiplex-nested RT-PCR offered sensitivities of 97.4% and 98.9% and specificities of 100% and 98.8% for diagnosing mature gametocytes of P. falciparum and P. vivax, respectively. The minimum detection limit of the multiplex-nested PCR was 10 copies of templates. Conclusions The multiplex-nested RT-PCR developed herein is useful for simultaneous assessment of both P. falciparum and P. vivax gametocyte carriage that is prevalent and generally sympatric in several malaria-endemic areas outside Africa. |
format |
Article in Journal/Newspaper |
author |
Kuamsab Napaporn Putaporntip Chaturong Pattanawong Urassaya Jongwutiwes Somchai |
author_facet |
Kuamsab Napaporn Putaporntip Chaturong Pattanawong Urassaya Jongwutiwes Somchai |
author_sort |
Kuamsab Napaporn |
title |
Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR |
title_short |
Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR |
title_full |
Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR |
title_fullStr |
Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR |
title_full_unstemmed |
Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR |
title_sort |
simultaneous detection of plasmodium vivax and plasmodium falciparum gametocytes in clinical isolates by multiplex-nested rt-pcr |
publisher |
BMC |
publishDate |
2012 |
url |
https://doi.org/10.1186/1475-2875-11-190 https://doaj.org/article/587755320969404ab3e7ca1ef1a93d44 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 11, Iss 1, p 190 (2012) |
op_relation |
http://www.malariajournal.com/content/11/1/190 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-190 1475-2875 https://doaj.org/article/587755320969404ab3e7ca1ef1a93d44 |
op_doi |
https://doi.org/10.1186/1475-2875-11-190 |
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Malaria Journal |
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11 |
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190 |
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1766348310942056448 |