Japanese Journal of Veterinary Research;Volume 66 Number 4

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Universal nematode detection by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) of purified nematode nucleic acids

Yoshino, Tomoo;Asakawa, Mitsuhiko;Osa, Yuichi;Kaneko, Masami;Endoh, Daiji

Permalink : http://hdl.handle.net/2115/72025
JaLCDOI : 10.14943/jjvr.66.4.311
KEYWORDS : degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR);inverted repeat;molecular typing PCR

Abstract

In this study, we have described a modified degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) method for molecular typing of nematodes collected from wild birds. To design the DOP-PCR, we selected 50 nonamers as DOP-motifs referring the data from a nematode genome. Inverted repeats of nonamers with a 100-1,500 base interval in the reported nematode genomes on the selection of the 50 nonamers. In these nonamers, 5 nonamers showed to create ladder pattern by DOP-PCR on 9, 6, 1, 2 and 3 species of morphologically identified adult nematodes, respectively. Eleven species of nematodes were distinguishable by combining the results of the two primers. It was suggested that the nematode species could be distinguished by DOP-PCR with a combination of 2 nonamers.

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