From: Plasmodium vivax molecular diagnostics in community surveys: pitfalls and solutions
Requirement for publication | Experimental information to be reported |
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Imperative | Sampling details of finger prick or venous blood sampling; e.g. type of filter paper (treated or not), microtainer/tubes (heparin, EDTA), storage solution (RNAprotect/Trizol) |
Description of extraction method; e.g. extracted blood volume, spin columns, chelex, type of DNAse treatment of purified RNA | |
Resuspension/elution volume for extracted DNA or RNA | |
Description of molecular target; e.g. Gene ID, amplicon size, primer and probe sequences | |
Reagent concentrations and total reaction volume; e.g. concentrations of primer/probe, template volume added to amplification reaction | |
Standard used for quantification; e.g. parasite trendline (with stage composition), NIBSC WHO reference standard, plasmid (linearized/supercoiled) | |
Clear definition of quantification results; e.g. method used for conversion of copy numbers into parasites/µL blood, clear denominator for “template copy number/µL whole blood or DNA” | |
Assay performance parameters; e.g. specificity, assay LOD, PCR efficiency | |
Optional | Storage conditions and time prior to extraction, e.g. of filter papers or whole blood, temperature, desiccant, particularly for filter papers |
Reproducibility; e.g. results from duplicates or triplicates | |
Comparison to LM; e.g. correlation of parasite counts/µL blood to copy numbers/µL blood |