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01.12.2012 | Methodology | Ausgabe 1/2012 Open Access

Malaria Journal 1/2012

Dried Plasmodium falciparum-infected samples as positive controls for malaria rapid diagnostic tests

Zeitschrift:
Malaria Journal > Ausgabe 1/2012
Autoren:
Michael Aidoo, Jaymin C Patel, John W Barnwell
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2875-11-239) contains supplementary material, which is available to authorized users.

Competing interests

The authors declare that they have no competing interests.

Authors' contributions

MA conceived and designed the study. MA and JP carried out the experiments. MA, JP and JWB analysed the data and wrote the manuscript. All authors read and approved the final manuscript.

Abstract

Background

Rapid diagnostic tests (RDTs) are central to fulfilling the WHO’s recommendation for parasitologic confirmation of all suspected cases of malaria. RDT performance may be compromised when exposed to the high temperature conditions typical of most malaria endemic regions. However, a systematic method to monitor RDT quality and performance in endemic countries is lacking at the present time. Current methods to monitor RDT performance in the field include comparing results from RDTs to diagnoses made by light microscopy and observing health workers perform tests. These methods are not substitutes for direct quality control. In this study, the suitability of dried Plasmodium falciparum-infected blood as quality control samples for malaria RDTs was evaluated.

Methods

Three cultured strains of P. falciparum at 200 and 2,000 parasites/μl were tested on 10 brands of RDT. After baseline testing to determine initial reactivity, aliquots of parasite-infected blood were air dried, stored at 35°C, room temperature (~25°C) or 4°C for one, four and 12 weeks and were then tested on the 10 RDTs after rehydration. Extended stability testing of dried blood stored at 4°C was done using P. falciparum strain 3D7 at 1,000 and 2,000 parasites/μl.

Results

All dried blood samples at 2,000 parasites/μl retained reactivity (100% sensitivity) at all three temperatures and time points for all nine RDT brands that detect histidine-rich protein-2 (HRP2). The dried blood samples with 200 parasites/μl were detected by six of the nine HRP2-based RDTs at all storage temperatures and time points. The sensitivity for two of the three remaining HRP2-based RDTs was 100% up to four weeks of storage at all temperatures but dropped to 87.5% at week 12. Of the four RDTs that detect plasmodium lactate dehydrogenase (pLDH) in a pan-specific manner, alone or in combination with HRP2, the detection of pLDH in samples with 2,000 parasites/μL was 100% for two RDTs and 80% for the other two RDTs. The mean level for detection of pLDH at 200 parasites/μl was low (29%), with a range of 0% to100%, which was partly attributable to weak initial baseline reactivity. Reactivity of dried 3D7 at 1,000 and 2,000 parasites/μl stored at 4°C was retained at 100% for up to 52 weeks for both HRP2 and pLDH.

Conclusions

In the absence of native or recombinant positive control antigens, well-standardized P. falciparum-infected dried blood samples can be used as positive control samples for monitoring RDT performance, particularly with HRP2-detecting tests.
Zusatzmaterial
Additional file 1 Supplementary information: Dried Tube Experiments. Colour-coded results for 10 RDT brands. Results are shown as relative band intensities for control and test bands. (PDF 207 kb) (PDF 208 KB)
12936_2012_2441_MOESM1_ESM.pdf
Additional file 2 Supplementary information: Punctured RDT Testing. Time series experiments using punctured RDT pouches. Test results for intact and punctured RDTs 2 and 5 stored at room temperature or 37°C and tested over time. (PDF 189 kb) (PDF 189 KB)
12936_2012_2441_MOESM2_ESM.pdf
Authors’ original file for figure 1
12936_2012_2441_MOESM3_ESM.pdf
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