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Quantification of Plasmodium ex vivo drug susceptibility by flow cytometry

Wirjanata, Grennady, Handayuni, Irene, Prayoga, Pak, Apriyanti, Dwi, Chalfein, Ferryanto, Sebayang, Boni F., Kho, Steven, Noviyanti, Rintis, Kenangalem, Enny, Campo, Brice, Poespoprodjo, Jeanne R., Price, Ric N. and Marfurt, Jutta (2015). Quantification of Plasmodium ex vivo drug susceptibility by flow cytometry. Malaria Journal,14(Article No. 417).

Document type: Journal Article
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IRMA ID 10444xPUB33
NHMRC Grant No. 1023438
Title Quantification of Plasmodium ex vivo drug susceptibility by flow cytometry
Author Wirjanata, Grennady
Handayuni, Irene
Prayoga, Pak
Apriyanti, Dwi
Chalfein, Ferryanto
Sebayang, Boni F.
Kho, Steven
Noviyanti, Rintis
Kenangalem, Enny
Campo, Brice
Poespoprodjo, Jeanne R.
Price, Ric N.
Marfurt, Jutta
Journal Name Malaria Journal
Publication Date 2015
Volume Number 14
Issue Number Article No. 417
ISSN 1475-2875   (check CDU catalogue  open catalogue search in new window)
Scopus ID 2-s2.0-84944908661
Total Pages 10
Place of Publication United Kingdom
Publisher BioMed Central Ltd.
HERDC Category C1 - Journal Article (DIISR)
Abstract Background
The emergence and spread of multidrug-resistant Plasmodium falciparum and Plasmodium vivax highlights the need for objective measures of ex vivo drug susceptibility. Flow cytometry (FC) has potential to provide a robust and rapid quantification of ex vivo parasite growth.

Methods

Field isolates from Papua, Indonesia, underwent ex vivo drug susceptibility testing against chloroquine, amodiaquine, piperaquine, mefloquine, and artesunate. A single nucleic acid stain (i.e., hydroethidine (HE) for P. falciparum and SYBR Green I (SG) for P. vivax) was used to quantify infected red blood cells by FC-based signal detection. Data derived by FC were compared to standard quantification by light microscopy (LM). A subset of isolates was used to compare single and double staining techniques.

Results
In total, 57 P. falciparum and 23 P. vivax field isolates were collected for ex vivo drug susceptibility testing. Reliable paired data between LM and FC was obtained for 88 % (295/334) of these assays. The median difference of derived IC50 values varied from −5.4 to 6.1 nM, associated with 0.83–1.23 fold change in IC50 values between LM and FC. In 15 assays (5.1 %), the derived difference of IC50 estimates was beyond the 95 % limits of agreement; in eleven assays (3.7 %), this was attributable to low parasite growth (final schizont count < 40 %), and in four assays (1.4 %) due to low initial parasitaemia at the start of assay (<2000 µl−1). In a subset of seven samples, LM, single and double staining FC techniques generated similar IC50 values.

Conclusions

A single staining FC-based assay using a portable cytometer provides a simple, fast and versatile platform for field surveillance of ex vivo drug susceptibility in clinical P. falciparum and P. vivax isolates.
Keywords Malaria
Plasmodium falciparum
Plasmodium vivax
Ex vivo drug susceptibility
Drug resistance
Flow cytometry
DOI http://dx.doi.org/10.1186/s12936-015-0940-8   (check subscription with CDU E-Gateway service for CDU Staff and Students  check subscription with CDU E-Gateway in new window)
Additional Notes This is an Open Access article distributed under the terms of the Creative Commons Attribution License 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Description for Link Link to CC Attribution 4.0 License
URL https://creativecommons.org/licenses/by/4.0/au


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