Real-time polymerase chain reaction for rapid detection of genes encoding SHV extended-spectrum β-lactamases.
OBJECTIVE
This study aimed to develop an improved method for the detection of bacterial SHV-type extended-spectrum β-lactamases (ESBLs).


METHODS
Our method was based on real-time polymerase chain reaction (PCR) in which the amplification of the product was monitored with a ﬂuorescent probe. This method enabled the detection of bla SHV genes with high degrees of sensitivity and speciﬁcity.


RESULTS
Based on ESBL phenotyping methods and bla gene DNA sequencing, we identified 240 bla genes from 662 Enterobacteriaceae isolated from clinical culture specimens. Of these 240 isolates, 26 had the bla SHV-28 genotype and three had the bla SHV-1 genotype. With our new real-time PCR assay, we detected 29 out of 29 bla SHV genes in ESBL-producing isolates.


CONCLUSIONS
This method represents a powerful tool for epidemiological studies of SHV ESBLs. Furthermore, it has potential for use in diagnostic microbiology.
Department of Pathology & Laboratory Medicine, Zayed Military Hospital, PO Box 3740, Abu Dhabi, United Arab Emirates. uaenow@eim.ae
M S
MS
Alfaresi
A A
AA
Elkoush
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004755:Enterobacteriaceae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D004756:Enterobacteriaceae Infections / Q000453:epidemiology / Q000382:microbiology; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D005456:Fluorescent Dyes; D005838:Genotype; D006801:Humans; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D008826:Microbial Sensitivity Tests; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013997:Time Factors; D001618:beta-Lactamases / Q000235:genetics*
D000900:Anti-Bacterial Agents; D005456:Fluorescent Dyes; C054882:beta-lactamase PIT-2; D001618:beta-Lactamases