PCR Primers for Trematode detection

Up to now a multitude of molecular markers have been developed for detection or identification of trematode species. Many base on the internal transcribed spacers of ribosomal DNA (ITS). These sequences are quite similar in the genome of related species, which makes it very easy to analyze them and design primers for distinguishing between certain species. Other attempts base on neutral tandem repeated sequences, that are not under selective pressure. They are present in very high copy numbers which offers the possibility to detect traces of DNA of a certain species. They also evolve more rapidly than ITS sequences, offering a greater chance to find species specific differences.
In cooperation with Joseph Hamburger (Haddash Medical School, Jerusalem, Israel) we started to look for tandem repeated sequences, that are suitable to design specific PCR primers for the detection of larval stages in snail and plankton samples. A specific detection will open avenues for ecological research on trematode larval stages in the field, where many organisms hinder microscopic examination. We already developed primers for Trichobilharzia spp. and E. caproni. Together with the primers of Hamburger et al. they are now employed for the analysis of samples from semi-filed experiments on the transmission and ecology of Trichobilharzia ocellata, Schistosoma mansoni and Echinostoma caproni miracidia.

Primers for the amplification of neutral tandem repeated sequences of trematodes.
Forward Primer    Reverse Primer    Species    Author   
GACGACAATAGTTGGGGTG GAATGGTATACGAGCTTGCC Echinostoma caproni HERTEL J. et al., 2003
CTGAATCTCTCGTTTGTTC GTTCCAGGTGAGTCTCTCTA Opisthorchis viverrini WONGRATANACHEEWIN S. et al., 2001
GATCTCACCTATCAGACGAAAC TCACAACGATACGATACGACCAAC Schistosoma haematobium HAMBURGER J. et al., 2001
GAAAATCGTTGTATCTCCG GGTGACCTGCCTAAAAATAC Schistosoma mansoni HAMBURGER J. et al., 1998
GTGACTTGCTACAGGTTGG GAATGGTATACGAGCTTGCC Trichobilharzia spp. HERTEL J. et al., 2002
GACGACAATAGTTGGGGTG GAATGGTATACGAGCTTGCC Trichobilharzia spp. HERTEL J. et al., 2002

On the job: Wilfried Haas, Jan Hertel.

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