Limit of detection of LAMP reactions using the boil and spin DNA extraction method.

Abstract

Representative amplifications experiments using serial dilutions of samples of known parasitaemia (quantified using the WHO International Standard as calibrator).Quantification was achieved by extracting DNA from a parasite dilution series made in whole blood using a Qiacube and DNA blood mini kit (Qiagen, De Hilden). Extracted DNA was then quantified using the Shokoples real time PCR [28] against a within run standard curve created from DNA extracted from the WHO international standard using the same methodology. Parasite count per microLitre was calculated given an initial parasite burden of 10% in the WHO standard and an assumed red cell count of 5 x106 red cells per microLitre. Mean time to turbidity in minutes as follows: 10,000 p/μL = 15.8; 1,000 p/μL = 15.9; 100 p/μL = 16.7; 10 p/μL = 18.2; 1 p/μL = 24.5.</p