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Polymerase chain reaction‐linked single‐strand conformation polymorphism of ribosomal DNA to fingerprint parasites
Author(s) -
Gasser Robin B.,
Monti Jennifer R.,
Zhu Xingquan,
Chilton Neil B.,
Hung GuoChiuan,
Guldberg Per
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150180913
Subject(s) - polymerase chain reaction , single strand conformation polymorphism , dna , ribosomal dna , biology , single strand , genetics , dna profiling , fingerprint (computing) , microbiology and biotechnology , computational biology , gene , computer science , phylogenetics , computer security
To overcome limitations of the morphological identification of some parasites (and their different developmental stages) to species, we have established a polymerase chain reaction‐linked single‐strand conformation polymorphism technique (PCR‐SSCP) utilizing the second internal transcribed spacer (ITS‐2) of ribosomal (r)DNA. This spacer was specifically chosen in the study of strongylid and ascarid nematodes because it is known to provide reliable species markers. The ITS‐2 from individual parasites was amplified by PCR, then denatured and subjected to electrophoresis on a mutation detection enhancement (MDE) (nondenaturing) gel matrix. PCR‐SSCP analysis showed that the single‐strand ITS‐2 patterns produced allowed the accurate identification of species. The method also allowed the display of (low‐level) variation in patterns within some species between different geographical isolates. These findings demonstrate the usefulness of PCR‐SSCP of ITS‐2 for the identification of nematode species and indicate its potential for resolving variation in the ITS‐2 sequence within species.