Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
2010-5-10
pubmed:abstractText
The second internal transcribed spacer (ITS2) of ribosomal DNA (rDNA) is used as a genetic marker to identify trichostrongylid nematodes. However, it is often difficult to amplify by polymerase chain reaction (PCR) the ITS2 rDNA of a single trichostrongylid nematode larva or egg. A nested PCR (nPCR) assay was, therefore, developed to amplify the ITS2 from individual trichostrongylid nematode larvae. The results show that the ITS2 rDNA of a significantly greater proportion of individual larvae was amplified using nPCR compared with a standard PCR. There was also no need to column-purify the genomic DNA before nPCR, which is more time and cost effective for studies involving large sample sizes. The amplicons produced from the secondary phase of the nPCR were subjected to single-strand conformation polymorphism analyses and DNA sequencing to confirm the species identity of the larvae used in the current study as Ostertagia gruehneri. The nPCR assay was also used to amplify the ITS2 from individual trichostrongylid eggs. The ability to amplify the ITS2 rDNA from large numbers of individual nematode eggs and larvae has important implications for diagnostic testing and for conducting epidemiological studies on these parasites of veterinary importance.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
1040-6387
pubmed:author
pubmed:issnType
Print
pubmed:volume
22
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
433-7
pubmed:meshHeading
pubmed-meshheading:20453221-Animals, pubmed-meshheading:20453221-Cattle, pubmed-meshheading:20453221-Cattle Diseases, pubmed-meshheading:20453221-DNA, Ribosomal, pubmed-meshheading:20453221-Female, pubmed-meshheading:20453221-Gene Amplification, pubmed-meshheading:20453221-Goat Diseases, pubmed-meshheading:20453221-Goats, pubmed-meshheading:20453221-Helminthiasis, pubmed-meshheading:20453221-Larva, pubmed-meshheading:20453221-Ostertagia, pubmed-meshheading:20453221-Ovum, pubmed-meshheading:20453221-Polymerase Chain Reaction, pubmed-meshheading:20453221-Polymorphism, Restriction Fragment Length, pubmed-meshheading:20453221-RNA, Ribosomal, pubmed-meshheading:20453221-Sheep, pubmed-meshheading:20453221-Sheep Diseases, pubmed-meshheading:20453221-Swine, pubmed-meshheading:20453221-Swine Diseases, pubmed-meshheading:20453221-Transcription, Genetic, pubmed-meshheading:20453221-Trichostrongyloidea
pubmed:year
2010
pubmed:articleTitle
Amplification of the second internal transcribed spacer ribosomal DNA of individual trichostrongylid nematode larvae by nested polymerase chain reaction.
pubmed:affiliation
Department of Biology, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E2, Canada. Canada.neil.chilton@usask.ca
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't