Development and validation of a multiplex PCR assay for identification of the epidemic ST-258/512 KPC-producing Klebsiella pneumoniae clone.
The Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-KP) sequence type (ST)-258/512 clone is the dominant clone by which KPC has disseminated worldwide. Standard typing methods are time-consuming and are therefore impractical for identification of this clone in the course of an outbreak. Through comparative genomic study, we have previously identified several presumably unique genes of this clone: 1) PILV-like protein (pilv-l), 2) transposase, IS66-family (is-66), and a 3) phage-related protein (prp). Our aims were to 1) test for the presence of these genes using a multiplex PCR in a large, multinational collection of KPC-KP isolates and to 2) validate this assay as a typing method for the identification of the ST-258/512 clone. KPC-KP isolates (n=160) that included both ST-258/512 (group A, n=114) and non-ST-258 (group B, n=46) strains were collected from the following countries: Greece, 20; Israel, 93; Italy, 19; USA, 25; and Colombia, 3. Group B included 30 different STs from various lineages. The pilv-l gene was present in 111/114 of ST-258 isolates, including all of the KPC-negative isolates resulting in a sensitivity of 97%. Using primers for a unique ST-258 pilv-l allele resulted in a specificity of 100%. The sensitivity values of is-66 and prp genes for detecting KPC-KP ST-258 were 83 and 89%, respectively, and the specificity values were 67 and 93%, respectively. PCR for the unique pilv-l ST-258 allele provides a reliable tool for rapid detection of the ST-258 clone. This method can be helpful both in the setting of an outbreak and in a large-scale survey of KPC-KP strains.
Division of Epidemiology, Tel-Aviv Sourasky Medical Center, Tel-Aviv, Israel. Electronic address: amosa@tasmc.health.gov.il.
Amos
A
Adler
Division of Epidemiology, Tel-Aviv Sourasky Medical Center, Tel-Aviv, Israel.
Efrat
E
Khabra
Division of Epidemiology, Tel-Aviv Sourasky Medical Center, Tel-Aviv, Israel.
Inna
I
Chmelnitsky
Department of Microbiology, National School of Public Health, Athens, Greece.
Panagiota
P
Giakkoupi
Department of Microbiology, National School of Public Health, Athens, Greece.
Alkiviadis
A
Vatopoulos
Division of Infectious Diseases and International Health, University of Virginia Health System, Charlottesville, VA, USA.
Amy J
AJ
Mathers
Division of Infectious Diseases and International Health, University of Virginia Health System, Charlottesville, VA, USA.
Anthony J
AJ
Yeh
Division of Infectious Diseases and International Health, University of Virginia Health System, Charlottesville, VA, USA.
Costi D
CD
Sifri
Division of Infectious Diseases, Università Cattolica Sacro Cuore, Rome, Italy.
Giulia
G
De Angelis
Division of Infectious Diseases, Università Cattolica Sacro Cuore, Rome, Italy; Department of Internal Medicine I, Medizinische Klinik, Universitätsklinikum Tübingen, Tübingen, Germany.
Evelina
E
Tacconelli
International Center for Medical Research and Training, Cali, Colombia.
Maria-Virginia
MV
Villegas
International Center for Medical Research and Training, Cali, Colombia.
John
J
Quinn
Division of Epidemiology, Tel-Aviv Sourasky Medical Center, Tel-Aviv, Israel.
Yehuda
Y
Carmeli
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007710:Klebsiella Infections / Q000382:microbiology*; D007711:Klebsiella pneumoniae / Q000145:classification* / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D017720:Molecular Epidemiology / Q000379:methods; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D001618:beta-Lactamases; C063912:carbapenemase
Epidemic clone; KPC; Typing