Detection of Plasmodium falciparum Infection with the Fluorescent Dye, Benzothiocarboxypurine

Mt, Makler; Lk, Ries; Ries, Judith; Rj, Horton; Dj, Hinrichs

doi:10.4269/ajtmh.1991.44.11

Cited by 33 publications

(18 citation statements)

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“…Various attempts have been made to improve the sensitivity and specificity of smear examination. All these and some other developments in diagnosis of malaria require sophisticated, high cost gadgets,modern infrastructure and specially trained manpower [5][6][7][8][9][10][11][12][13]. Due to this rider, these developments have mostly remained confined to research laboratories or to a few diagnostic centres.…”

Section: Discussionmentioning

confidence: 99%

Saving Private Ryan: The Indian Scenario (Rapid Diagnosis of Malaria at Regimental Aid Post)

Gokhale

2004

Medical Journal Armed Forces India

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Immunochromatography test (ICT) (Paracheck Pf) for diagnosis of Plasmodium falciparum (Pf) infection was compared with the conventional smear examination method. A total of 350 specimens of blood from cases of fever were investigated (falciparum malaria 220, vivax malaria 100, controls 30). Paracheck Pf ICT was found to have enormous advantages over smear examination due to its high degree of sensitivity, specificity, speed and ease of performance. Paracheck Pf ICT test kits are stable at room temperature. Regimental medical officers (RMOs) and nursing assistants with minimal training can safely practise Paracheck Pf ICT method. Introduction of this test method in the Armed Forces can facilitate early diagnosis and specific treatment of falciparum malaria even at far flung places. This will have enormous beneficial effect in reducing morbidity due to malaria and saving precious lives. In short as well as long term, it is a viable cost effective option. MJAFI 2004; 60 : 137-141

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Section: Discussionmentioning

confidence: 99%

Saving Private Ryan: The Indian Scenario (Rapid Diagnosis of Malaria at Regimental Aid Post)

Gokhale

2004

Medical Journal Armed Forces India

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“…An alternative fluorochrome procedure uses a solution of BCP (7,8,40). BCP can be applied directly to a lysed blood suspension or to an unfixed but dry thick blood film and stains the nucleic acid of viable P. falciparum parasites intensely.…”

Section: Diagnosis Using Fluorescence Microscopymentioning

confidence: 99%

Rapid Diagnostic Tests for Malaria Parasites

2002

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SUMMARY Malaria presents a diagnostic challenge to laboratories in most countries. Endemic malaria, population movements, and travelers all contribute to presenting the laboratory with diagnostic problems for which it may have little expertise available. Drug resistance and genetic variation has altered many accepted morphological appearances of malaria species, and new technology has given an opportunity to review available procedures. Concurrently the World Health Organization has opened a dialogue with scientists, clinicians, and manufacturers on the realistic possibilities for developing accurate, sensitive, and cost-effective rapid diagnostic tests for malaria, capable of detecting 100 parasites/μl from all species and with a semiquantitative measurement for monitoring successful drug treatment. New technology has to be compared with an accepted Rgold standardS that makes comparisons of sensitivity and specificity between different methods. The majority of malaria is found in countries where cost-effectiveness is an important factor and ease of performance and training is a major consideration. Most new technology for malaria diagnosis incorporates immunochromatographic capture procedures, with conjugated monoclonal antibodies providing the indicator of infection. Preferred targeted antigens are those which are abundant in all asexual and sexual stages of the parasite and are currently centered on detection of HRP-2 from Plasmodium falciparum and parasite-specific lactate dehydrogenase or Plasmodium aldolase from the parasite glycolytic pathway found in all species. Clinical studies allow effective comparisons between different formats, and the reality of nonmicroscopic diagnoses of malaria is considered.

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“…The principle behind the assay is the contrast between host erythrocytes, which lack DNA and RNA, and the malaria parasites, which do not and which are thus readily stained with dyes that show enhanced fluorescence in the presence of nucleic acids (8). Fluorescence-based in vitro antimalarial assays have been developed previously (1,12) but have required complex, multistep protocols or additional equipment not amenable to rapid, high-throughput use.…”

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Simple and Inexpensive Fluorescence-Based Technique for High-Throughput Antimalarial Drug Screening

Smilkstein

Sriwilaijaroen

Kelly³

et al. 2004

Antimicrob Agents Chemother

1,039

945

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Radioisotopic assays involve expense, multistep protocols, equipment, and radioactivity safety requirements which are problematic in high-throughput drug testing. This study reports an alternative, simple, robust, inexpensive, one-step fluorescence assay for use in antimalarial drug screening. Parasite growth is determined by using SYBR Green I, a dye with marked fluorescence enhancement upon contact with Plasmodium DNA. A side-by-side comparison of this fluorescence assay and a standard radioisotopic method was performed by testing known antimalarial agents against Plasmodium falciparum strain D6. Both assay methods were used to determine the effective concentration of drug that resulted in a 50% reduction in the observed counts (EC 50 ) after 48 h of parasite growth in the presence of each drug. The EC 50 s of chloroquine, quinine, mefloquine, artemisinin, and 3,6-bis--(N,N-diethylamino)-amyloxyxanthone were similar or identical by both techniques. The results obtained with this new fluorescence assay suggest that it may be an ideal method for highthroughput antimalarial drug screening.The global scope of malaria and the spread of drug-resistant Plasmodium falciparum make the need for improved therapy undeniable (4). Assessment of both existing drugs and new antimalarials, alone or in combination, requires reliable methods for high-throughput testing. For decades, antimalarial drug effects have been measured in vitro by quantifying parasite uptake of radioactive substrates as a measure of growth and viability in the presence of the test drug (2, 3). [3 H]hypoxanthine is the most widely used radiolabel, but because it requires purine starvation prior to the assay, the [ 3 H]ethanolamine assay is favored by our laboratory. While these methods are accurate and reliable, they rely on relatively expensive radioisotopes and multistep procedures that become increasingly problematic and impractical as the volume of testing increases. We report on the development of an alternative, fluorescencebased in vitro assay for use for the high-throughput screening of the activities of drugs against P. falciparum. The goals for the assay included simplicity, cost savings, robust performance, applicability to automated analysis, and speed.The principle behind the assay is the contrast between host erythrocytes, which lack DNA and RNA, and the malaria parasites, which do not and which are thus readily stained with dyes that show enhanced fluorescence in the presence of nucleic acids (8). Fluorescence-based in vitro antimalarial assays have been developed previously (1, 12) but have required complex, multistep protocols or additional equipment not amenable to rapid, high-throughput use. This report establishes and validates a facile, one-step method that uses the same principle. MATERIALS AND METHODSChloroquine diphosphate, quinine sulfate, artemisinin, and saponin were purchased from Sigma Aldrich Chemical Company (St. Louis, Mo.); mefloquine was obtained from Walter Reed Army Institute for Research (Silver Spring, Md.); and 3,6-b...

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Detection of Plasmodium falciparum Infection with the Fluorescent Dye, Benzothiocarboxypurine

Cited by 33 publications

References 0 publications

Saving Private Ryan: The Indian Scenario (Rapid Diagnosis of Malaria at Regimental Aid Post)

Saving Private Ryan: The Indian Scenario (Rapid Diagnosis of Malaria at Regimental Aid Post)

Rapid Diagnostic Tests for Malaria Parasites

Simple and Inexpensive Fluorescence-Based Technique for High-Throughput Antimalarial Drug Screening

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