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Journal of Clinical Microbiology, November 2000, p. 4072-4075, Vol. 38, No. 11
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Detection and Quantification of Plasmodium falciparum in Blood Samples Using Quantitative Nucleic Acid Sequence-Based Amplification

Gerard J. Schoone,1 Linda Oskam,1,* Nel C. M. Kroon,1 Henk D. F. H. Schallig,1 and Sabah A. Omar2

Royal Tropical Institute, Department of Biomedical Research, 1105 AZ Amsterdam, The Netherlands,1 and Kenya Medical Research Institute, Centre for Biotechnology, Nairobi, Kenya2

Received 3 February 2000/Returned for modification 4 April 2000/Accepted 22 August 2000

A quantitative nucleic acid sequence-based amplification (QT-NASBA) assay for the detection of Plasmodium parasites has been developed. Primers and probes were selected on the basis of the sequence of the small-subunit rRNA gene. Quantification was achieved by coamplification of the RNA in the sample with one modified in vitro RNA as a competitor in a single-tube NASBA reaction. Parasite densities ranging from 10 to 108 Plasmodium falciparum parasites per ml could be demonstrated and quantified in whole blood. This is approximately 1,000 times more sensitive than conventional microscopy analysis of thick blood smears. Comparison of the parasite densities obtained by microscopy and QT-NASBA with 120 blood samples from Kenyan patients with clinical malaria revealed that for 112 of 120 (93%) of the samples results were within a 1-log difference. QT-NASBA may be especially useful for the detection of low parasite levels in patients with early-stage malaria and for the monitoring of the efficacy of drug treatment.

* Corresponding author. Mailing address: Royal Tropical Institute (KIT), Department of Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands. Phone: 31-20-5665441. Fax: 31-20-6971841. E-mail: L.Oskam{at}kit.nl.

Journal of Clinical Microbiology, November 2000, p. 4072-4075, Vol. 38, No. 11
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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