Publication: High-throughput ultrasensitive molecular techniques for quantifying low-density malaria parasitemias.
Accepted Date
2014-06-23
Issued Date
2014-09
Copyright Date
2014
Resource Type
Language
eng
ISSN
0095-1137 (printed)
1098-660X (electronic)
1098-660X (electronic)
Rights
Mahidol University
Rights Holder(s)
PubMed Central
Bibliographic Citation
Imwong M, Hanchana S, Malleret B, Rénia L, Day NP, Dondorp A. et al. High-throughput ultrasensitive molecular techniques for quantifying low-density malaria parasitemias. J Clin Microbiol. 2014 Sep;52(9):3303-9.
Suggested Citation
Mallika Imwong, มัลลิกา อิ่มวงศ์, Sarun Hanchana, Malleret, Benoit, Rénia, Laurent, Day, Nicholas P. J., Dondorp, Arjen, Nosten, Francois, Snounou, Georges, White, Nicholas J. High-throughput ultrasensitive molecular techniques for quantifying low-density malaria parasitemias.. Imwong M, Hanchana S, Malleret B, Rénia L, Day NP, Dondorp A. et al. High-throughput ultrasensitive molecular techniques for quantifying low-density malaria parasitemias. J Clin Microbiol. 2014 Sep;52(9):3303-9.. doi:10.1128/JCM.01057-14. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/848
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Title
High-throughput ultrasensitive molecular techniques for quantifying low-density malaria parasitemias.
Corresponding Author(s)
Other Contributor(s)
Mahidol University. Faculty of Tropical Medicine. Department of Molecular Tropical Medicine and Genetics.
Mahidol University. Faculty of Tropical Medicine. Mahidol Oxford Research Unit.
Mahidol University. Faculty of Tropical Medicine. Shoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit.
Mahidol University. Faculty of Tropical Medicine. Mahidol Oxford Research Unit.
Mahidol University. Faculty of Tropical Medicine. Shoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit.
Abstract
The epidemiology of malaria in "low-transmission" areas has been underestimated.
Molecular detection methods have revealed higher prevalences of malaria than
conventional microscopy or rapid diagnostic tests, but these typically evaluate
finger-prick capillary blood samples (∼5 μl) and therefore cannot detect parasite
densities of <200/ml. Their use underestimates true parasite carriage rates. To
characterize the epidemiology of malaria in low-transmission settings and plan
elimination strategies, more sensitive quantitative PCR (qPCR) is needed to
identify and quantify low-density malaria parasitemias. A highly sensitive
"high-volume" quantitative PCR (qPCR) method based on Plasmodium sp. 18S RNA was
adapted for blood sample volumes of ≥250 μl and scaled for high throughput. The
methods were validated by assessment of the analytical sensitivity and
specificity, diagnostic sensitivity, and specificity, efficiency, precision,
analytical and diagnostic accuracies, limit of detection, root cause analysis of
false positives, and robustness. The high-volume qPCR method based on Plasmodium
sp. 18S RNA gave high PCR efficiency of 90 to 105%. Concentrations of parasite
DNA from large volumes of blood gave a consistent analytical detection limit
(LOD) of 22 parasites/ml (95% CI, 21.79 to 74.9), which is some 2,500 times more
sensitive than conventional microscopy and 50 times more sensitive than currently
used PCR methods from filter paper blood spots. The diagnostic specificity was
99.75%. Using automated procedures it was possible to process 700 blood samples
per week. A very sensitive and specific high-throughput high-volume qPCR method
for the detection of low-density parasitemias (>20 parasites/ml) was developed
and validated.