Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2003-2-7
pubmed:abstractText
We developed and optimized a new modified amplified fragment length polymorphism (AFLP) typing method to obtain a multibanding fingerprint that can be separated by agarose gel electrophoresis. Both to maximize the discriminatory power and to facilitate the computer-assisted analysis, bacterial DNA was digested with four different restriction enzymes. After ligation of adaptors to the DNA fragments, PCR testing of various single primers was performed. Two single primers that gave optimal results with regard to band resolution and discriminatory power were selected and combined. The computer-assisted analysis of fingerprint patterns was performed with Pearson's product-moment correlation values of densitometric curves, without assigning bands to peaks. Thus, the analysis is not subject to human interpretation errors. With this method, we investigated two outbreaks of multiresistant Klebsiella pneumoniae in an intensive care unit and various sporadic isolates of K. pneumoniae and Klebsiella oxytoca. Cluster analysis of isolates analyzed in different experiments and on different gels showed that fingerprint patterns clustered correctly according to subspecies or to the outbreaks. Multienzyme multiplex PCR AFLP revealed that the first outbreak was caused by two different types of strains. Outbreak two was caused by yet another strain of K. pneumoniae. In conclusion, the typing method used here is easy to perform and highly reproducible, and due to generation of complex banding patterns, it has a higher discriminatory power. Furthermore, the multienzyme multiplex PCR fingerprints are easy to analyze, and a reliable database can be stored in the computer to facilitate comparison of future isolates of Klebsiella spp. The method can be performed in every clinical microbiology laboratory.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-10488158, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-10523569, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-10584467, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11015382, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11049703, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11117641, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11117642, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11121492, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11161899, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11209841, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-11526149, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-7501463, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-7665634, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-8880485, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-8880498, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-9630406, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-9749400, http://linkedlifedata.com/resource/pubmed/commentcorrection/12574286-9767057
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0095-1137
pubmed:author
pubmed:issnType
Print
pubmed:volume
41
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
798-802
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2003
pubmed:articleTitle
Use of multienzyme multiplex PCR amplified fragment length polymorphism typing in analysis of outbreaks of multiresistant Klebsiella pneumoniae in an intensive care unit.
pubmed:affiliation
Laboratory of Medical Microbiology, St Elisabeth Hospital, Tilburg, The Netherlands. a.vd.zee@elisabeth.nl
pubmed:publicationType
Journal Article