Development and validation of a multiplex PCR assay for identification of the epidemic ST-258/512 KPC-producing Klebsiella pneumoniae clone

Amos Adler*, Efrat Khabra, Inna Chmelnitsky, Panagiota Giakkoupi, Alkiviadis Vatopoulos, Amy J. Mathers, Anthony J. Yeh, Costi D. Sifri, Giulia De Angelis, Evelina Tacconelli, Maria Virginia Villegas, John Quinn, Yehuda Carmeli

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Pages (from-to)12-15
Number of pages4
JournalDiagnostic Microbiology and Infectious Disease
Issue number1
StatePublished - Jan 2014
Externally publishedYes


FundersFunder number
Seventh Framework Programme241796
European Commission


    • Epidemic clone
    • KPC
    • Typing


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