Differentiation of Entamoeba histolytica and Entamoeba dispar Cysts Using Polymerase Chain Reaction on DNA Isolated from Faeces with Spin Columns |
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Authors: | J J Verweij J Blotkamp E A T Brienen A Aguirre A M Polderman |
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Institution: | (1) Department of Parasitology, Leiden University Medical Centre, PO Box 9600, 2300 RC Leiden, The Netherlands e-mail: j.j.verweij@lumc.nl, NL;(2) Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK, GB |
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Abstract: | Since Entamoeba histolytica and Entamoeba dispar were formally recognized as two different species at the World Health Organization (WHO)/Pan American Health Organization
(PAHO)/United Nations Educational, Scientific and Cultural Organization (UNESCO) meeting in Mexico City in 1997, the specific
differentiation of the two morphologically identical species would seem relevant in clinical diagnosis. Several polymerase
chain reaction (PCR)-based methods have been described and used successfully, but methods for DNA isolation from cysts in
stool samples are time-consuming and problematic due to inhibitory factors in faeces. The use of the slightly modified QIAamp
tissue method (Qiagen, Germany) for DNA isolation was evaluated in 657 unpreserved faecal samples from cases of suspected Entamoeba histolytica/Entamoeba dispar infection. In only 1.7% of the cases was PCR hampered by inhibitors present in the faeces. The DNA isolation procedure was
found to be rapid, simple and one that could easily be implemented in a routine diagnostic setting. In 98.8% of Entamoeba histolytica/Entamoeba dispar cyst-positive faecal samples, the true identity of the cysts could be determined using PCR specific for Entamoeba histolytica and Entamoeba dispar, respectively. |
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