Comparison of two real-time PCR assays for the detection of malaria parasites from hemolytic blood samples – Short communication

There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

Abstract

We compared the performance of an in-house and a commercial malaria polymerase chain reaction (PCR) assay using freeze–thawed hemolytic blood samples.

A total of 116 freeze–thawed ethylenediamine tetraacetic acid (EDTA) blood samples of patients with suspicion of malaria were analyzed by an in-house as well as by a commercially available real-time PCR.

Concordant malaria negative PCR results were reported for 39 samples and malaria-positive PCR results for 67 samples. The in-house assay further detected one case of Plasmodium falciparum infection, which was negative in the commercial assay as well as five cases of P. falciparum malaria and three cases of Plasmodium vivax malaria, which showed sample inhibition in the commercial assay. The commercial malaria assay was positive in spite of a negative in-house PCR result in one case. In all concordant results, cycle threshold values of P. falciparum-positive samples were lower in the commercial PCR than in the in-house assay.

Although Ct values of the commercial PCR kit suggest higher sensitivity in case of concordant results, it is prone to inhibition if it is applied to hemolytic freeze–thawed blood samples. The number of misidentifications was, however, identical for both real-time PCR assays.

Related collections

Most cited references 17

Record: found
Abstract: found
Article: found

Is Open Access

Assessment of the prozone effect in malaria rapid diagnostic tests

Philippe Gillet, Marcella Mori, Marjan Van Esbroeck … (2009)

Background The prozone effect (or high doses-hook phenomenon) consists of false-negative or false-low results in immunological tests, due to an excess of either antigens or antibodies. Although frequently cited as a cause of false-negative results in malaria rapid diagnostic tests (RDTs), especially at high parasite densities of Plasmodium falciparum, it has been poorly documented. In this study, a panel of malaria RDTs was challenged with clinical samples with P. falciparum hyperparasitaemia (> 5% infected red blood cells). Methods Twenty-two RDT brands were tested with seven samples, both undiluted and upon 10 ×, 50 × and 100 × dilutions in NaCl 0.9%. The P. falciparum targets included histidine-rich protein-2 (HRP-2, n = 17) and P. falciparum-specific parasite lactate dehydrogenase (Pf-pLDH, n = 5). Test lines intensities were recorded in the following categories: negative, faint, weak, medium or strong. The prozone effect was defined as an increase in test line intensity of at least one category after dilution, if observed upon duplicate testing and by two readers. Results Sixteen of the 17 HRP-2 based RDTs were affected by prozone: the prozone effect was observed in at least one RDT sample/brand combination for 16/17 HRP-2 based RDTs in 6/7 samples, but not for any of the Pf-pLDH tests. The HRP-2 line intensities of the undiluted sample/brand combinations with prozone effect (n = 51) included a single negative (1.9%) and 29 faint and weak readings (56.9%). The other target lens (P. vivax-pLDH, pan-specific pLDH and aldolase) did not show a prozone effect. Conclusion This study confirms the prozone effect as a cause of false-negative HRP-2 RDTs in samples with hyperparasitaemia.

0 comments Cited 52 times – based on 0 reviews      Review now

Bookmark

Record: found
Abstract: found
Article: not found

How useful is PCR in the diagnosis of malaria?

Thomas Hänscheid, Martin P Grobusch (2002)

Polymerase chain reaction (PCR) assays are the most sensitive and specific method to detect malaria parasites, and have acknowledged value in research settings. However, the time lag between sample collection, transportation and processing, and dissemination of results back to the physician limits the usefulness of PCR in routine clinical practice. Furthermore, in most areas with malaria transmission, factors such as limited financial resources, persistent subclinical parasitaemia, inadequate laboratory infrastructures in the poorer, remote rural areas preclude PCR as a diagnostic method. Even in affluent, non-endemic countries, PCR is not a suitable method for routine use. Nonetheless, PCR could be clinically useful in selected situations.

0 comments Cited 51 times – based on 0 reviews      Review now

Bookmark

Record: found
Abstract: found
Article: not found

Microfluidic approaches to malaria detection.

Mathuros Ruchirawat, Peter R. C. Gascoyne, JUTAMAAD SATAYAVIVAD (2004)

Microfluidic systems are under development to address a variety of medical problems. Key advantages of micrototal analysis systems based on microfluidic technology are the promise of small size and the integration of sample handling and measurement functions within a single, automated device having low mass-production costs. Here, we review the spectrum of methods currently used to detect malaria, consider their advantages and disadvantages, and discuss their adaptability towards integration into small, automated micro total analysis systems. Molecular amplification methods emerge as leading candidates for chip-based systems because they offer extremely high sensitivity, the ability to recognize malaria species and strain, and they will be adaptable to the detection of new genotypic signatures that will emerge from current genomic-based research of the disease. Current approaches to the development of chip-based molecular amplification are considered with special emphasis on flow-through PCR, and we present for the first time the method of malaria specimen preparation by dielectrophoretic field-flow-fractionation. Although many challenges must be addressed to realize a micrototal analysis system for malaria diagnosis, it is concluded that the potential benefits of the approach are well worth pursuing.

0 comments Cited 31 times – based on 0 reviews      Review now

Bookmark

All references

Author and article information

Journal

Journal ID (nlm-ta): Eur J Microbiol Immunol (Bp)

Journal ID (iso-abbrev): Eur J Microbiol Immunol (Bp)

Journal ID (publisher-id): EUJMI

Title: European Journal of Microbiology & Immunology

Publisher: Akadémiai Kiadó (Budapest )

ISSN (Print): 2062-509X

ISSN (Electronic): 2062-8633

Publication date (Electronic): 18 June 2015

Publication date Collection: June 2015

Volume: 5

Issue: 2

Pages: 159-163

Affiliations

[1 ]Department of Tropical Medicine at the Bernhard Nocht Institute, German Armed Forces Hospital of Hamburg , Germany

[2 ]Bernhard Nocht Institute for Tropical Medicine Hamburg , Germany

[3 ]Institute for Microbiology, Virology and Hygiene, University Medicine Rostock , Germany

Author notes

* Department of Tropical Medicine at the Bernhard Nocht Institute, German Armed Forces Hospital of Hamburg, Bernhard Nocht street 74, D-20359 Hamburg, Germany; 004940/694728743; 004940/694728709; Frickmann@ 123456bni-hamburg.de

Article

DOI: 10.1556/1886.2015.00006

PMC ID: 4500067

License:

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Comparison of two real-time PCR assays for the detection of malaria parasites from hemolytic blood samples – Short communication

Read this article at

Abstract

Related collections

European Journal of Microbiology and Immunology

Most cited references 17

Assessment of the prozone effect in malaria rapid diagnostic tests

How useful is PCR in the diagnosis of malaria?

Microfluidic approaches to malaria detection.

Author and article information

Journal

Affiliations

Author notes

Article

Page count

Categories

Comments

Comment on this article

Similar content 140

Cited by 4

Most referenced authors 132