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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
The best practice for preparation of samples from FTA
®
cards for diagnosis of blood borne infections using African trypanosomes as a model system
Parasites and Vectors, Volume 4, No. 1, Article 68, Year 2011
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Description
Background: Diagnosis of blood borne infectious diseases relies primarily on the detection of the causative agent in the blood sample. Molecular techniques offer sensitive and specific tools for this although considerable difficulties exist when using these approaches in the field environment. In large scale epidemiological studies, FTA®cards are becoming increasingly popular for the rapid collection and archiving of a large number of samples. However, there are some difficulties in the downstream processing of these cards which is essential for the accurate diagnosis of infection. Here we describe recommendations for the best practice approach for sample processing from FTA®cards for the molecular diagnosis of trypanosomiasis using PCR. Results: A comparison of five techniques was made. Detection from directly applied whole blood was less sensitive (35.6%) than whole blood which was subsequently eluted from the cards using Chelex®100 (56.4%). Better apparent sensitivity was achieved when blood was lysed prior to application on the FTA cards (73.3%) although this was not significant. This did not improve with subsequent elution using Chelex®100 (73.3%) and was not significantly different from direct DNA extraction from blood in the field (68.3%). Conclusions: Based on these results, the degree of effort required for each of these techniques and the difficulty of DNA extraction under field conditions, we recommend that blood is transferred onto FTA cards whole followed by elution in Chelex®100 as the best approach. © 2011 Ahmed et al; licensee BioMed Central Ltd.
Authors & Co-Authors
Ahmed, Heba A.
United Kingdom, Edinburgh
University of Edinburgh, College of Medicine and Veterinary Medicine
Egypt, Zagazig
Faculty of Veterinary Medicine
MacLeod, Ewan Thomas
United Kingdom, Edinburgh
University of Edinburgh, College of Medicine and Veterinary Medicine
Hide, Geoff
United Kingdom, Salford
University of Salford
Welburn, Susan Christina
United Kingdom, Edinburgh
University of Edinburgh, College of Medicine and Veterinary Medicine
Picozzi, Kim
United Kingdom, Edinburgh
University of Edinburgh, College of Medicine and Veterinary Medicine
Statistics
Citations: 89
Authors: 5
Affiliations: 3
Identifiers
Doi:
10.1186/1756-3305-4-68
e-ISSN:
17563305
Research Areas
Genetics And Genomics