RRC ID |
15958
|
著者 |
Fujita S, Yosizaki K, Ogushi T, Uechi K, Takemori Y, Senda Y.
|
タイトル |
Rapid identification of gram-negative bacteria with and without CTX-M extended-spectrum β-lactamase from positive blood culture bottles by PCR followed by microchip gel electrophoresis.
|
ジャーナル |
J Clin Microbiol
|
Abstract |
We evaluated the usefulness of PCR analysis of the 16S-23S rRNA gene internal transcribed spacer (ITS) and the CTX-M extended-spectrum β-lactamase (ESBL) followed by microchip gel electrophoresis (MGE) for direct identification and CTX-M detection of Gram-negative bacteria (GNB) from positive blood culture bottles. Of 251 GNB isolated from blood cultures containing a single bacterium, 225 (90%) were correctly identified at the species level directly from positive blood culture bottles by comparing the ITS-PCR patterns of the sample strain with those of the control strains. There were no cases of incorrect identification. Limitations encountered included the inability to detect mixed cultures (four bottles) as well as some species (Enterobacter species and Klebsiella oxytoca) demonstrating identical ITS-PCR patterns. A total of 109 ESBL-producing isolates from various clinical materials obtained between January 2005 and December 2008 were examined for bla(CTX-M), bla(SHV), and bla(TEM) genes by PCR and sequences of PCR products. CTX-M ESBL was detected in 105 isolates, and SHV ESBL was detected in two isolates. The remaining two isolates (K. oxytoca) were shown to harbor bla(OXY.) Twenty (19%) of 104 Escherichia coli isolates from blood cultures were suspected to produce ESBL by the combination disk method, and these isolates were shown to harbor CTX-M ESBL by PCR-MGE. The results were obtained within 1.5 h at a calculated cost of $6.50 per specimen. In conclusion, simultaneous detection of ITS length polymorphisms and bla(CTX)-(M) by single PCR followed by MGE is useful for rapid, cost-effective, and reliable species-level identification of CTX-M ESBL-producing GNB responsible for bloodstream infections.
|
巻・号 |
49(4)
|
ページ |
1483-8
|
公開日 |
2011-4-1
|
DOI |
10.1128/JCM.01976-10
|
PII |
JCM.01976-10
|
PMID |
21289149
|
PMC |
PMC3122828
|
MeSH |
Bacteremia / diagnosis*
Bacteremia / microbiology
Bacteriological Techniques / methods*
Blood / microbiology*
DNA, Bacterial / genetics
DNA, Ribosomal Spacer / genetics
Electrophoresis / methods*
Gram-Negative Bacteria / genetics
Gram-Negative Bacteria / isolation & purification*
Gram-Negative Bacterial Infections / diagnosis*
Gram-Negative Bacterial Infections / microbiology
Humans
Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Time Factors
beta-Lactamases / genetics
|
IF |
5.897
|
引用数 |
20
|
WOS 分野
|
MICROBIOLOGY
|
リソース情報 |
一般微生物 |
JCM 1027
JCM 1090
JCM 1233
JCM 1236
JCM 1239
JCM 1277
JCM 1474
JCM 1656
JCM 1668
JCM 1669
JCM 1672
JCM 1675
JCM 1673
JCM 1685
JCM 1975
JCM 1976
JCM 3725
JCM 5490
JCM 5833
JCM 5963
JCM 5967
JCM 5968
JCM 5969
JCM 6156
JCM 6158
JCM 6840
JCM 6841
JCM 6842
JCM 9659 |