A Radioimmunoassay for the Diagnosis of Malaria

Avidor, Boaz; Golenser, Jacob; Schutte, C. H. J.; Cox, G. A.; Isaäcson, M; Sulitzeanu, D.

doi:10.4269/ajtmh.1987.37.225

Cited by 8 publications

(4 citation statements)

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“…At a higher level of health services, accurate and objective parasite counts are needed to carefully monitor malaria infection in the follow-up of treatment of, for instance, drug-resistant parasites. However, the required sensitive detection of malaria parasites especially in large numbers of blood samples still poses problems (22).In the last few years, alternatives to microscopic detection of malaria parasites in thick or thin blood smears have been investigated, such as immunological methods to demonstrate antibodies or antigens (1,17,18), detection of parasites by fluorescence micros copy (24,25), and the use of malaria-specific radioactively labeled DNA and RNA probes (4). Microscopy is still widely considered to be the gold standard when comparing alternative methods, but its performance is highly influenced by the local circumstances and working conditions and on the availability of expertise, 'This research was financed in part by the Netherlands' Ministry for Development Co-operation (project ID.…”

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“…In the last few years, alternatives to microscopic detection of malaria parasites in thick or thin blood smears have been investigated, such as immunological methods to demonstrate antibodies or antigens (1,17,18), detection of parasites by fluorescence microscopy (24,25), and the use of malaria-specific radioactively labeled DNA and RNA probes (4). Microscopy is still widely considered to be the gold standard when comparing alternative methods, but its performance is highly influenced by the local circumstances and working conditions and on the availability of expertise, ' which is especially a problem in the field (2,3).…”

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Flow cytometric screening of blood samples for malaria parasites

et al. 1993

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method for the detection and estimation of malaria parasites in blood samples using flow cytometry is presented. In a single-step procedure 50 ~1 of blood sample was collected in 1 ml of lysis solution containing formaldehyde, causing red blood cells to lyse while parasites and white blood cells are preserved. Thus prepared, samples could be transported and remained stored in lysis solution until flow cytometric analysis was performed. The cells were stained for DNA with the fluorescent dye Hoechst 33258 and subsequently analyzed by a FACStar flow cytometer. Parasites and white blood cells were distinguished and counted based on blue Hoechst fluorescence and forward scattering. Since red blood cells were lysed, parasite numbers were given related to the number of white blood cells similar to what is done in microscopic examination of thick blood smears. In dilution experiments with animal and human material, parasite counts by flow cytometry correlated very well with the theoretically calculated numbers (regression coefficients of > 0.94). In human material parasitemias of -0.005% were detected. In a pilot study, 700 samples were collected in Thailand and screened by microscopic examination of thick smears and by flow cytometry; 29 were found positive by combining both methods, 2 were missed by flow cytometry, and 20 were missed by microscopists in the field. After microscopic reexamination in the central laboratory, 15 of these 20 were found positive, 5 remained unconfirmed. Key terms: Plasmodium, Hoechst 33258, epidemiological surveysThe slow progress in the development of antimalarial vaccines and the rapidly spreading resistance to commonly used and newly developed drugs reemphasize the need to control malaria at the level of primary health care. Efficient control of malaria requires sensitive and quantitative techniques for the screening of relatively large numbers of people for the presence of malaria parasites. Epidemiological studies covering large areas are currently being performed to monitor the spread of drug resistance, or for the follow-up of drug trials and in the future to follow up vaccine campaigns. At a higher level of health services, accurate and objective parasite counts are needed to carefully monitor malaria infection in the follow-up of treatment of, for instance, drug-resistant parasites. However, the required sensitive detection of malaria parasites especially in large numbers of blood samples still poses problems (22).In the last few years, alternatives to microscopic detection of malaria parasites in thick or thin blood smears have been investigated, such as immunological methods to demonstrate antibodies or antigens (1,17,18), detection of parasites by fluorescence microscopy (24,25), and the use of malaria-specific radioactively labeled DNA and RNA probes (4). Microscopy is still widely considered to be the gold standard when comparing alternative methods, but its performance is highly influenced by the local circumstances and working conditions and on the availability of expertise...

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Flow cytometric screening of blood samples for malaria parasites

et al. 1993

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“…Several promising approaches to the development of improved malaria diagnosis are under evaluation, including the quantitative buffy coat procedure (15), use of antigen-targeted antibodies (1), and oligonucleotide hybridization which targets repetitive DNA (4) or rRNA (16). We have previously demonstrated the feasibility of using synthetic DNA (9, 10,12) and enzyme-linked synthetic DNA (7,11,13) to detect repetitive DNA nonisotopically.…”

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Optimization of a rapid nonisotopic DNA probe assay for Plasmodium falciparum in the Gambia

et al. 1991

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An enzyme-linked synthetic DNA probe which hybridizes to repetitive DNA of Plasmodium fakciparum was used in conjunction with a microtiter-based lysis and filtration blood processing procedure. An assay protocol was developed that is more sensitive and robust than previous protocols, which use stored blood and phenol extraction. In comparison with thick smear examination, 33% positive, 60% negative, and 7% conflicting

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“…Hutchinson et al developed a simplified radioimmunoassay for diagnostic serology [15]. Later this technique was constantly used for the diagnosis of malaria [16], congenital adrenal hyperplasia [17] endocrine tumors of pancreas [18]. Lately, emergence of fluorescent immunocapture techniques with parallel sensitivity and safety superposed the radioimmunoassays.…”

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