The design and evaluation of a shaped filter collection device to sample and store defined volume dried blood spots from finger pricks

Malaria Journal

Methodology

James Oliver¹  Frank Tully¹  Spencer D Polley²  Peter L Chiodini³  Iveth J González⁴  Mark D Perkins⁴  David Bell⁵ 

[1] 42 Technology Limited, Meadow Lane, PE27 4LG, St. Ives, Cambridgeshire, UK;Department of Clinical Parasitology, Hospital for Tropical Diseases, Mortimer Market, Capper St, University College London NHS Foundation Trust, WC1E 6JB, London, UK;Department of Clinical Parasitology, Hospital for Tropical Diseases, Mortimer Market, Capper St, University College London NHS Foundation Trust, WC1E 6JB, London, UK;London School of Hygiene & Tropical Medicine, Keppel Street, WC1E 7HT, London, UK;FIND (Foundation for Innovative New Diagnostics), Campus Biotech, 9 Chemin des Mines, 1202, Geneva, Switzerland;Global Good, Intellectual Ventures Lab, 1555 132nd Ave NE, 98005, Bellevue, WA, USA;

关键词: Dried blood spot;    Malaria;    Sampling;    Shaped filter paper;    DNA extraction;    PCR;    LAMP;   

DOI  :  10.1186/s12936-015-0558-x

 received in 2014-09-17, accepted in 2015-01-10,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundDried blood spots are a common medium for collecting patient blood prior to testing for malaria by molecular methods. A new shaped filter device for the quick and simple collection of a designated volume of patient blood has been designed and tested against conventional blood spots for accuracy and precision.MethodsShaped filter devices were laser cut from Whatman GB003 paper to absorb a 20 μl blood volume. These devices were used to sample Plasmodium falciparum infected blood and the volume absorbed was measured volumetrically. Conventional blood spots were made by pipetting 20 μl of the same blood onto Whatman 3MM paper. DNA was extracted from both types of dried blood spot using Qiagen DNA blood mini or Chelex extraction for real-time PCR analysis, and PURE extraction for malaria LAMP testing.ResultsThe shaped filter devices collected a mean volume of 21.1 μl of blood, with a coefficient of variance of 8.1%. When used for DNA extraction by Chelex and Qiagen methodologies the mean number of international standard units of P. falciparum DNA recovered per μl of the eluate was 53.1 (95% CI: 49.4 to 56.7) and 32.7 (95% CI: 28.8 to 36.6), respectively for the shaped filter device, and 54.6 (95% CI: 52.1 to 57.1) and 12.0 (95% CI: 9.9 to 14.1), respectively for the 3MM blood spots. Qiagen extraction of 200 μl of whole infected blood yielded 853.6 international standard units of P. falciparum DNA per μl of eluate.ConclusionsA shaped filter device provides a simple way to quickly sample and store a defined volume of blood without the need for any additional measuring devices. Resultant dried blood spots may be employed for DNA extraction using a variety of technologies for nucleic acid amplification without the need for repeated cleaning of scissors or punches to prevent cross contamination of samples and results are comparable to traditional DBS.

【 授权许可】

CC BY   
© Polley et al.; licensee BioMed Central. 2015