Establishment of the 1st World Health Organization International Standard for Plasmodium falciparum DNA for nucleic acid amplification technique (NAT)-based assays.
BACKGROUND
In order to harmonize results for the detection and quantification of Plasmodium falciparum DNA by nucleic acid amplification technique (NAT)-based assays, a World Health Organization (WHO) collaborative study was performed, evaluating a series of candidate standard preparations.


METHODS
Fourteen laboratories from 10 different countries participated in the collaborative study. Four candidate preparations based upon blood samples parasitaemic for P. falciparum were evaluated in the study. Sample AA was lyophilized, whilst samples BB, CC and DD were liquid/frozen preparations. The candidate standards were tested by each laboratory at a range of dilutions in four independent assays, using both qualitative and quantitative NAT-based assays. The results were collated and analysed statistically.


RESULTS
Twenty sets of data were returned from the participating laboratories and used to determine the mean P. falciparum DNA content for each sample. The mean log10 "equivalents"/ml were 8.51 for sample AA, 8.45 for sample BB, 8.35 for sample CC, and 5.51 for sample DD. The freeze-dried preparation AA, was examined by accelerated thermal degradation studies and found to be highly stable.


CONCLUSIONS
On the basis of the collaborative study, the freeze-dried material, AA (NIBSC code No. 04/176) was established as the 1st WHO International Standard for P. falciparum DNA NAT-based assays and has been assigned a potency of 10(9) International Units (IU) per ml. Each vial contains 5 x 10(8) IU, equivalent to 0.5 ml of material after reconstitution.
Padley
David J
DJ
National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. dpadley@nibsc.ac.uk
Heath
Alan B
AB
Sutherland
Colin
C
Chiodini
Peter L
PL
Baylis
Sally A
SA
D000818:Animals; D001681:Biological Assay / Q000379:methods / Q000592:standards; D019411:Clinical Laboratory Techniques / Q000592:standards; D016054:DNA, Protozoan / Q000235:genetics*; D007391:International Cooperation; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000592:standards*; D010963:Plasmodium falciparum / Q000235:genetics*; D014944:World Health Organization
D016054:DNA, Protozoan