DETECTION, IDENTIFICATION AND SEMI-QUANTIFICATION OF MALARIA PARASITES BY NASBA AMPLIFICATION OF SMALL-SUBUNIT RIBOSOMAL-RNA SEQUENCES

We have adapted the Nucleic Acid Sequence Based Amplification method ( NASBA), a molecular detection method for the amplification of specific RNA sequences, for the detection, identification and semi-quantificat ion of malaria parasites. Primers and probes were selected based on th e nucleotide sequence of the small subunit ribosomal RNA gene. NASBA e nabled the detection of as little as 0.01 parasitized erythrocytes per mu l blood. When applied to blood samples of patients, NASBA allowed the detection of malaria parasites in confirmed malaria patients. in a ddition, malaria parasites were detected in a number of patients with a history of suspected malaria. Semi-quantification over a wide parasi te range was achieved by analysing serial end-point dilutions of RNA p reparations, Semi-quantitative NASBA analysis of malaria parasites in follow-up samples of treated malaria patients showed a rapid decline o f parasite levels in most patients and a low level of persistent paras ites in some of the patients. (C) 1997 Elsevier Science B.V. All right s reserved.