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01.12.2012 | Research | Ausgabe 1/2012 Open Access

Malaria Journal 1/2012

Investigation of volatile organic biomarkers derived from Plasmodium falciparum in vitro

Zeitschrift:
Malaria Journal > Ausgabe 1/2012
Autoren:
Rina PM Wong, Gavin R Flematti, Timothy ME Davis
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2875-11-314) contains supplementary material, which is available to authorized users.

Competing interests

The author(s) declare that they have no competing interests.

Authors’ contributions

RW participated in study design, experimentation, data analysis and drafted the manuscript. GF participated in study design, GC-MS optimization, data analysis and critical revision of the manuscript. TD conceived the study, and participated in its design and coordination, and critical revision of the manuscript. All authors read and approved the final manuscript.

Abstract

Background

There remains a need for techniques that improve the sensitive detection of viable Plasmodium falciparum as part of diagnosis and therapeutic monitoring in clinical studies and usual-care management of malaria infections. A non-invasive breath test based on P. falciparum- associated specific volatile organic compounds (VOCs) could fill this gap and provide insights into parasite metabolism and pathogenicity. The aim of this study was to determine whether VOCs are present in the headspace above in vitro P. falciparum cultures.

Methods

A novel, custom-designed apparatus was developed to enable efficient headspace sampling of infected and non-infected cultures. Conditions were optimized to support cultures of high parasitaemia (>20%) to improve the potential detection of parasite-specific VOCs. A number of techniques for VOC analysis were investigated including solid phase micro-extraction using two different polarity fibres, and purge and trap/thermal desorption, each coupled to gas chromatography–mass spectrometry. Each experiment and analysis method was performed at least on two occasions. VOCs were identified by comparing their mass spectra against commercial mass spectral libraries.

Results

No unique malarial-specific VOCs could be detected relative to those in the control red blood cell cultures. This could reflect sequestration of VOCs into cell membranes and/or culture media but solvent extractions of supernatants and cell lysates using hexane, dichloromethane and ethyl acetate also showed no obvious difference compared to control non-parasitized cultures.

Conclusions

Future in vivo studies analysing the breath of patients with severe malaria who are harbouring a parasite biomass that is significantly greater than achievable in vitro may yet reveal specific clinically-useful volatile chemical biomarkers.
Zusatzmaterial
Authors’ original file for figure 1
12936_2012_2208_MOESM1_ESM.tiff
Authors’ original file for figure 2
12936_2012_2208_MOESM2_ESM.tiff
Literatur
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