Development of a stable positive control to be used for quality assurance of rapid diagnostic tests for malaria.
The objective of this study is to develop and evaluate a simple, cheap, and stable positive control for the quality control and quality assurance (QA) of rapid diagnostic tests (RDT) for the diagnosis of malaria. Plasmodium falciparum in vitro culture of known parasite concentrations was dried on a protein saver card, that is, dried blood spots (DBSs). The cards were stored at temperatures ranging from 27 to 60 degrees C from 1 day up to 6 months. Antigens were subsequently eluted from the card giving final concentrations ranging from 30 000 parasites to 300 parasites/microL and tested for stability against RDT based on the antigens parasite lactate dehydrogenase (pLDH), aldolase, and histidine-rich protein 2 (HRP-2). HRP-2 antigens were stable throughout the whole study and yielded positive results irrespective of parasite concentration, storage duration, or temperature, although band intensity differences could be observed when high parasites were compared with low parasite densities. Aldolase was able to generate positive signals for up to 4 weeks irrespective of the storage conditions. Thereafter, intensities decreased proportionally to increasing temperature and storage duration. Thirty thousand parasites per liter could give a signal up to 16 weeks when stored at a temperature of maximum 45 degrees C. However, densities of 300 parasites/microL were not able to generate a signal during the study. pLDH, the least stable of the 3 antigens, was not able to generate a signal after 1 week of storage. The DBS method yields a very stable positive control for quality control and QA of RDTs based on HRP-2. RDTs based on aldolase may also benefit from this method although to a lesser extent because that particular antigen is less stable in the DBS system.
Koninklijk Instituut voor de Tropen/Royal Tropical Institute, KIT Biomedical Research, Amsterdam, The Netherlands.
Inge
I
Versteeg
Petra F
PF
Mens
D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis; D003955:Diagnostic Tests, Routine / Q000592:standards*; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D016778:Malaria, Falciparum / Q000175:diagnosis*; D010963:Plasmodium falciparum / Q000737:chemistry*; D055550:Protein Stability; D015800:Protozoan Proteins / Q000032:analysis; D011785:Quality Assurance, Health Care / Q000379:methods; D012015:Reference Standards; D013696:Temperature; D013997:Time Factors
D000953:Antigens, Protozoan; C514457:HRP-2 antigen, Plasmodium falciparum; D015800:Protozoan Proteins; D007770:L-Lactate Dehydrogenase; D005634:Fructose-Bisphosphate Aldolase