Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification
Abstract Background Malaria remains a global public health problem responsible for 445,000 deaths in 2016. While microscopy remains the mainstay of malaria diagnosis, highly sensitive molecular methods for detection of low-grade sub-microscopic infections are needed for surveillance studies and iden...
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ftdoajarticles:oai:doaj.org/article:41870688d7194c008ae627a3e7d5548c 2023-05-15T15:09:37+02:00 Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification Bryan Grabias Edward Essuman Isabella A. Quakyi Sanjai Kumar 2019-04-01T00:00:00Z https://doi.org/10.1186/s12936-019-2743-9 https://doaj.org/article/41870688d7194c008ae627a3e7d5548c EN eng BMC http://link.springer.com/article/10.1186/s12936-019-2743-9 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2743-9 1475-2875 https://doaj.org/article/41870688d7194c008ae627a3e7d5548c Malaria Journal, Vol 18, Iss 1, Pp 1-9 (2019) Plasmodium falciparum erythrocyte membrane protein 1 Parasitemia Real-time PCR Genomic DNA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2019 ftdoajarticles https://doi.org/10.1186/s12936-019-2743-9 2022-12-31T08:40:01Z Abstract Background Malaria remains a global public health problem responsible for 445,000 deaths in 2016. While microscopy remains the mainstay of malaria diagnosis, highly sensitive molecular methods for detection of low-grade sub-microscopic infections are needed for surveillance studies and identifying asymptomatic reservoirs of malaria transmission. Methods The Plasmodium falciparum genome sequence was analysed to identify high copy number genes that improve P. falciparum parasite detection in blood by RT-PCR. Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1)-specific primers were evaluated for P. falciparum detection in hospital-based microscopically positive dried blood spots and field-acquired whole blood from asymptomatic individuals from Ghana. Results PfEMP1 outperformed the Pf18S sequence for amplification-based P. falciparum detection. PfEMP1 primers exhibited sevenfold higher sensitivity compared to Pf18S primers for parasite genomic DNA. Probit analysis established a 95% detection threshold of 9.3 parasites/mL for PfEMP1 compared to 98.2 parasites/mL for Pf18S primers. The PfEMP1 primers also demonstrated superior clinical sensitivity, identifying 100% (20/20) of dried blood spot samples and 70% (69/98) of asymptomatic individuals as positive versus 55% (11/20) and 54% (53/98), respectively, for Pf18S amplification. Conclusions These results establish PfEMP1 as a novel amplification target for highly sensitive detection of both acute infections from filter paper samples and submicroscopic asymptomatic low-grade infections. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 18 1 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Plasmodium falciparum erythrocyte membrane protein 1 Parasitemia Real-time PCR Genomic DNA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Plasmodium falciparum erythrocyte membrane protein 1 Parasitemia Real-time PCR Genomic DNA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Bryan Grabias Edward Essuman Isabella A. Quakyi Sanjai Kumar Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification |
topic_facet |
Plasmodium falciparum erythrocyte membrane protein 1 Parasitemia Real-time PCR Genomic DNA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Malaria remains a global public health problem responsible for 445,000 deaths in 2016. While microscopy remains the mainstay of malaria diagnosis, highly sensitive molecular methods for detection of low-grade sub-microscopic infections are needed for surveillance studies and identifying asymptomatic reservoirs of malaria transmission. Methods The Plasmodium falciparum genome sequence was analysed to identify high copy number genes that improve P. falciparum parasite detection in blood by RT-PCR. Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1)-specific primers were evaluated for P. falciparum detection in hospital-based microscopically positive dried blood spots and field-acquired whole blood from asymptomatic individuals from Ghana. Results PfEMP1 outperformed the Pf18S sequence for amplification-based P. falciparum detection. PfEMP1 primers exhibited sevenfold higher sensitivity compared to Pf18S primers for parasite genomic DNA. Probit analysis established a 95% detection threshold of 9.3 parasites/mL for PfEMP1 compared to 98.2 parasites/mL for Pf18S primers. The PfEMP1 primers also demonstrated superior clinical sensitivity, identifying 100% (20/20) of dried blood spot samples and 70% (69/98) of asymptomatic individuals as positive versus 55% (11/20) and 54% (53/98), respectively, for Pf18S amplification. Conclusions These results establish PfEMP1 as a novel amplification target for highly sensitive detection of both acute infections from filter paper samples and submicroscopic asymptomatic low-grade infections. |
format |
Article in Journal/Newspaper |
author |
Bryan Grabias Edward Essuman Isabella A. Quakyi Sanjai Kumar |
author_facet |
Bryan Grabias Edward Essuman Isabella A. Quakyi Sanjai Kumar |
author_sort |
Bryan Grabias |
title |
Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification |
title_short |
Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification |
title_full |
Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification |
title_fullStr |
Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification |
title_full_unstemmed |
Sensitive real-time PCR detection of Plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification |
title_sort |
sensitive real-time pcr detection of plasmodium falciparum parasites in whole blood by erythrocyte membrane protein 1 gene amplification |
publisher |
BMC |
publishDate |
2019 |
url |
https://doi.org/10.1186/s12936-019-2743-9 https://doaj.org/article/41870688d7194c008ae627a3e7d5548c |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 18, Iss 1, Pp 1-9 (2019) |
op_relation |
http://link.springer.com/article/10.1186/s12936-019-2743-9 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2743-9 1475-2875 https://doaj.org/article/41870688d7194c008ae627a3e7d5548c |
op_doi |
https://doi.org/10.1186/s12936-019-2743-9 |
container_title |
Malaria Journal |
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18 |
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1 |
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1766340774367068160 |