Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples
Abstract Background The emergence of Plasmodium knowlesi in humans, which is in many cases misdiagnosed by microscopy as Plasmodium malariae due to the morphological similarity has contributed to the needs of detection and differentiation of malaria parasites. At present, nested PCR targeted on Plas...
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ftdoajarticles:oai:doaj.org/article:51044ec51edf4927b928527594ffeb23 2023-05-15T15:13:02+02:00 Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples Anthony Claudia N Chin Lit-Chein Cheong Fei-Wen Palaeya Vanitha Chang Phooi-Yee Mahmud Rohela Fong Mun-Yik Lau Yee-Ling Al-Mekhlafi Abdulsalam M Chen Yeng 2011-07-01T00:00:00Z https://doi.org/10.1186/1475-2875-10-197 https://doaj.org/article/51044ec51edf4927b928527594ffeb23 EN eng BMC http://www.malariajournal.com/content/10/1/197 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-10-197 1475-2875 https://doaj.org/article/51044ec51edf4927b928527594ffeb23 Malaria Journal, Vol 10, Iss 1, p 197 (2011) Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2011 ftdoajarticles https://doi.org/10.1186/1475-2875-10-197 2022-12-31T02:34:52Z Abstract Background The emergence of Plasmodium knowlesi in humans, which is in many cases misdiagnosed by microscopy as Plasmodium malariae due to the morphological similarity has contributed to the needs of detection and differentiation of malaria parasites. At present, nested PCR targeted on Plasmodium ssrRNA genes has been described as the most sensitive and specific method for Plasmodium detection. However, this method is costly and requires trained personnel for its implementation. Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method was developed for the clinical detection of P. knowlesi . The sensitivity and specificity of LAMP was evaluated in comparison to the results obtained via microscopic examination and nested PCR. Methods LAMP assay was developed based on P. knowlesi genetic material targeting the apical membrane antigen-1 (AMA-1) gene. The method uses six primers that recognize eight regions of the target DNA and it amplifies DNA within an hour under isothermal conditions (65°C) in a water-bath. Results LAMP is highly sensitive with the detection limit as low as ten copies for AMA-1. LAMP detected malaria parasites in all confirm cases (n = 13) of P. knowlesi infection (sensitivity, 100%) and none of the negative samples (specificity, 100%) within an hour. LAMP demonstrated higher sensitivity compared to nested PCR by successfully detecting a sample with very low parasitaemia (< 0.01%). Conclusion With continuous efforts in the optimization of this assay, LAMP may provide a simple and reliable test for detecting P. knowlesi malaria parasites in areas where malaria is prevalent. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 10 1 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Anthony Claudia N Chin Lit-Chein Cheong Fei-Wen Palaeya Vanitha Chang Phooi-Yee Mahmud Rohela Fong Mun-Yik Lau Yee-Ling Al-Mekhlafi Abdulsalam M Chen Yeng Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background The emergence of Plasmodium knowlesi in humans, which is in many cases misdiagnosed by microscopy as Plasmodium malariae due to the morphological similarity has contributed to the needs of detection and differentiation of malaria parasites. At present, nested PCR targeted on Plasmodium ssrRNA genes has been described as the most sensitive and specific method for Plasmodium detection. However, this method is costly and requires trained personnel for its implementation. Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method was developed for the clinical detection of P. knowlesi . The sensitivity and specificity of LAMP was evaluated in comparison to the results obtained via microscopic examination and nested PCR. Methods LAMP assay was developed based on P. knowlesi genetic material targeting the apical membrane antigen-1 (AMA-1) gene. The method uses six primers that recognize eight regions of the target DNA and it amplifies DNA within an hour under isothermal conditions (65°C) in a water-bath. Results LAMP is highly sensitive with the detection limit as low as ten copies for AMA-1. LAMP detected malaria parasites in all confirm cases (n = 13) of P. knowlesi infection (sensitivity, 100%) and none of the negative samples (specificity, 100%) within an hour. LAMP demonstrated higher sensitivity compared to nested PCR by successfully detecting a sample with very low parasitaemia (< 0.01%). Conclusion With continuous efforts in the optimization of this assay, LAMP may provide a simple and reliable test for detecting P. knowlesi malaria parasites in areas where malaria is prevalent. |
format |
Article in Journal/Newspaper |
author |
Anthony Claudia N Chin Lit-Chein Cheong Fei-Wen Palaeya Vanitha Chang Phooi-Yee Mahmud Rohela Fong Mun-Yik Lau Yee-Ling Al-Mekhlafi Abdulsalam M Chen Yeng |
author_facet |
Anthony Claudia N Chin Lit-Chein Cheong Fei-Wen Palaeya Vanitha Chang Phooi-Yee Mahmud Rohela Fong Mun-Yik Lau Yee-Ling Al-Mekhlafi Abdulsalam M Chen Yeng |
author_sort |
Anthony Claudia N |
title |
Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples |
title_short |
Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples |
title_full |
Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples |
title_fullStr |
Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples |
title_full_unstemmed |
Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples |
title_sort |
specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (lamp) in blood samples |
publisher |
BMC |
publishDate |
2011 |
url |
https://doi.org/10.1186/1475-2875-10-197 https://doaj.org/article/51044ec51edf4927b928527594ffeb23 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 10, Iss 1, p 197 (2011) |
op_relation |
http://www.malariajournal.com/content/10/1/197 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-10-197 1475-2875 https://doaj.org/article/51044ec51edf4927b928527594ffeb23 |
op_doi |
https://doi.org/10.1186/1475-2875-10-197 |
container_title |
Malaria Journal |
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10 |
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1 |
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1766343640020418560 |