Linked Life Data

11874894

Source:http://linkedlifedata.com/resource/pubmed/id/11874894

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2002-3-4
pubmed:abstractText
Two PCR protocols targeting the 18S rRNA gene of Cryptococcus neoformans were established, compared, and evaluated in murine cryptococcal meningitis. One protocol was designed as a nested PCR to be performed in conventional block thermal cyclers. The other protocol was designed as a quantitative single-round PCR adapted to LightCycler technology. One hundred brain homogenates and dilutions originating from 20 ICR mice treated with different azoles were examined. A fungal burden of 3 x 10(1) to 2.9 x 10(4) CFU per mg of brain tissue was determined by quantitative culture. Specific PCR products were amplified by the conventional and the LightCycler methods in 86 and 87 samples, respectively, with products identified by DNA sequencing and real-time fluorescence detection. An analytical sensitivity of 1 CFU of C. neoformans per mg of brain tissue and less than 10 CFU per volume used for extraction was observed for both PCR protocols, while homogenates of 70 organs from mice infected with other fungi were PCR negative. Specificity testing was performed with genomic DNA from 31 hymenomycetous fungal species and from the ustilaginomycetous yeast Malassezia furfur, which are phylogenetically related to C. neoformans. Twenty-four strains, including species of human skin flora like M. furfur and Trichosporon spp., were PCR negative. Amplification was observed with Cryptococcus amylolentus, Filobasidiella depauperata, Cryptococcus laurentii, and five species unrelated to clinical specimens. LightCycler PCR products from F. depauperata and Trichosporon faecale could be clearly discriminated by melting curve analysis. The sensitive and specific nested PCR assay as well as the rapid and quantitative LightCycler PCR assay might be useful for the diagnosis and monitoring of human cryptococcal infections.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-10655350, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-10747127, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-10770733, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-10843082, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-10921955, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-10937778, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-10948820, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-11283078, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-11283097, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-7650214, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-8126194, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-8873884, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-8897191, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-9145879, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-9738078, http://linkedlifedata.com/resource/pubmed/commentcorrection/11874894-9774618
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
1071-412X
pubmed:author
pubmed:issnType
Print
pubmed:volume
9
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
461-9
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2002
pubmed:articleTitle
Detection of Cryptococcus neoformans DNA in tissue samples by nested and real-time PCR assays.
pubmed:affiliation
Institute for Tropical Medicine, University Hospital Tübingen, Germany. ralf.bialek@med.uni-tuebingen.de
pubmed:publicationType
Journal Article