Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum

Abstract Background Rapid diagnostic tests (RDTs) that detect histidine-rich protein 2 (PfHRP2) are used throughout Africa for the diagnosis of Plasmodium falciparum malaria. However, recent reports indicate that parasites lacking the pfhrp2 and/or histidine-rich protein 3 (pfhrp3) genes, which prod...

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Main Authors:	Jonathan B. Parr, Olivia Anderson, Jonathan J. Juliano, Steven R. Meshnick
Format:	Article
Language:	English
Published:	BioMed Central 2018-04-01
Series:	Malaria Journal
Subjects:	Rapid diagnostic tests False-negative Diagnostic resistance Histidine-rich protein hrp2 hrp3
Online Access:	http://link.springer.com/article/10.1186/s12936-018-2287-4

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spelling	doaj-art-70c5f187213b48a190e3306fdf95e5912018-08-20T17:07:36ZengBioMed CentralMalaria Journal1475-28752018-04-011711810.1186/s12936-018-2287-4Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparumJonathan B. Parr0Olivia Anderson1Jonathan J. Juliano2Steven R. Meshnick3Division of Infectious Diseases, Department of Medicine, University of North CarolinaDepartment of Epidemiology, Gillings School of Global Public Health, University of North CarolinaDivision of Infectious Diseases, Department of Medicine, University of North CarolinaDivision of Infectious Diseases, Department of Medicine, University of North CarolinaAbstract Background Rapid diagnostic tests (RDTs) that detect histidine-rich protein 2 (PfHRP2) are used throughout Africa for the diagnosis of Plasmodium falciparum malaria. However, recent reports indicate that parasites lacking the pfhrp2 and/or histidine-rich protein 3 (pfhrp3) genes, which produce antigens detected by these RDTs, are common in select regions of South America, Asia, and Africa. Proving the absence of a gene is challenging, and multiple PCR assays targeting these genes have been described. A detailed characterization and comparison of published assays is needed to facilitate robust and streamlined testing approaches. Results Among six pfhrp2 and pfhrp3 PCR assays tested, the lower limit of detection ranged from 0.01 pg/µL to 0.1 ng/µL of P. falciparum 3D7 strain DNA, or approximately 0.4–4000 parasite genomes/µL. By lowering the elongation temperature to 60 °C, a tenfold improvement in the limit of detection and/or darker bands for all exon 1 targets and for the first-round reaction of a single exon 2 target was achieved. Additionally, assays targeting exon 1 of either gene yielded spurious amplification of the paralogous gene. Using these data, an optimized testing algorithm for the detection of pfhrp2- and pfhrp3-negative P. falciparum is proposed. Conclusions Surveillance of pfhrp2- and pfhrp3-negative P. falciparum requires careful laboratory workflows. PCR-based testing methods coupled with microscopy and/or antigen testing serve as useful tools to support policy development. Standardized approaches to the detection of pfhrp2- and pfhrp3-negative P. falciparum should inform efforts to define the impact of these parasites.http://link.springer.com/article/10.1186/s12936-018-2287-4Rapid diagnostic testsFalse-negativeDiagnostic resistanceHistidine-rich proteinhrp2hrp3
institution	Open Data Bank
collection	Open Access Journals
building	Directory of Open Access Journals
language	English
format	Article
author	Jonathan B. Parr Olivia Anderson Jonathan J. Juliano Steven R. Meshnick
spellingShingle	Jonathan B. Parr Olivia Anderson Jonathan J. Juliano Steven R. Meshnick Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum Malaria Journal Rapid diagnostic tests False-negative Diagnostic resistance Histidine-rich protein hrp2 hrp3
author_facet	Jonathan B. Parr Olivia Anderson Jonathan J. Juliano Steven R. Meshnick
author_sort	Jonathan B. Parr
title	Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum
title_short	Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum
title_full	Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum
title_fullStr	Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum
title_full_unstemmed	Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum
title_sort	streamlined, pcr-based testing for pfhrp2- and pfhrp3-negative plasmodium falciparum
publisher	BioMed Central
series	Malaria Journal
issn	1475-2875
publishDate	2018-04-01
description	Abstract Background Rapid diagnostic tests (RDTs) that detect histidine-rich protein 2 (PfHRP2) are used throughout Africa for the diagnosis of Plasmodium falciparum malaria. However, recent reports indicate that parasites lacking the pfhrp2 and/or histidine-rich protein 3 (pfhrp3) genes, which produce antigens detected by these RDTs, are common in select regions of South America, Asia, and Africa. Proving the absence of a gene is challenging, and multiple PCR assays targeting these genes have been described. A detailed characterization and comparison of published assays is needed to facilitate robust and streamlined testing approaches. Results Among six pfhrp2 and pfhrp3 PCR assays tested, the lower limit of detection ranged from 0.01 pg/µL to 0.1 ng/µL of P. falciparum 3D7 strain DNA, or approximately 0.4–4000 parasite genomes/µL. By lowering the elongation temperature to 60 °C, a tenfold improvement in the limit of detection and/or darker bands for all exon 1 targets and for the first-round reaction of a single exon 2 target was achieved. Additionally, assays targeting exon 1 of either gene yielded spurious amplification of the paralogous gene. Using these data, an optimized testing algorithm for the detection of pfhrp2- and pfhrp3-negative P. falciparum is proposed. Conclusions Surveillance of pfhrp2- and pfhrp3-negative P. falciparum requires careful laboratory workflows. PCR-based testing methods coupled with microscopy and/or antigen testing serve as useful tools to support policy development. Standardized approaches to the detection of pfhrp2- and pfhrp3-negative P. falciparum should inform efforts to define the impact of these parasites.
topic	Rapid diagnostic tests False-negative Diagnostic resistance Histidine-rich protein hrp2 hrp3
url	http://link.springer.com/article/10.1186/s12936-018-2287-4
_version_	1612685424284663808

Streamlined, PCR-based testing for pfhrp2- and pfhrp3-negative Plasmodium falciparum

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