The use of Multiple Displacement Amplification to increase the detection and genotyping of Trypanosoma samples immobilised on FTA filters.

Morrison, L.J., McCormack, G., Sweeney, L., Likeufack, A.C.L., Truc, P., Turner, C.M.R., Tait, A. and MacLeod, A. (2007) The use of Multiple Displacement Amplification to increase the detection and genotyping of Trypanosoma samples immobilised on FTA filters. American Journal of Tropical Medicine and Hygiene, 76(6), pp. 1132-1137.

Abstract

Whole genome amplification methods are a recently developed tool for amplifying DNA from limited template. We report its application in trypanosome infections, characterized by low parasitemias. Multiple displacement amplification (MDA) amplifies DNA with a simple in vitro step and was evaluated on mouse blood samples on FTA filter cards with known numbers of Trypanosoma brucei parasites. The data showed a 20-fold increase in the number of PCRs possible per sample, using primers diagnostic for the multicopy ribosomal ITS region or 177-bp repeats, and a 20-fold increase in sensitivity over nested PCR against a single-copy microsatellite. Using MDA for microsatellite genotyping caused allele dropout at low DNA concentrations, which was overcome by pooling multiple MDA reactions. The validity of using MDA was established with samples from Human African Trypanosomiasis patients. The use of MDA allows maximal use of finite DNA samples and may prove a valuable tool in studies where multiple reactions are necessary, such as population genetic analyses.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: MacLeod, Professor Annette and Morrison, Dr Lesley
• Authors: Morrison, L.J., McCormack, G., Sweeney, L., Likeufack, A.C.L., Truc, P., Turner, C.M.R., Tait, A., and MacLeod, A.
• Subjects: Q Science > QR Microbiology > QR355 Virology
• College/School: College of Medical Veterinary and Life Sciences; College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: American Journal of Tropical Medicine and Hygiene
• Publisher: American Society of Tropical Medicine and Hygiene
• ISSN: 0002-9637
• Copyright Holders: Copyright © 2007 American Society of Tropical Medicine and Hygiene
• First Published: First published in American Journal of Tropical Medicine and Hygiene 76(6):1132-1137
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher