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Volume 19, Number 6—June 2013

Synopsis

New Delhi Metallo-β-Lactamase–producing Enterobacteriaceae, United States

J. Kamile RasheedComments to Author , Brandon Kitchel, Wenming Zhu, Karen F. Anderson, Nancye C. Clark, Mary Jane Ferraro, Patrice Savard, Romney M. Humphries, Alexander J. Kallen, and Brandi M. Limbago
Author affiliations: Centers for Disease Control and Prevention, Atlanta, Georgia, USA (J.K. Rasheed, B. Kitchel, W. Zhu, K.F. Anderson, N.C. Clark, A.J. Kallen, B.M. Limbago); Massachusetts General Hospital, Boston, Massachusetts, USA (M.J. Ferraro); Johns Hopkins University, Baltimore, Maryland, USA (P. Savard); Johns Hopkins Health System, Baltimore (P. Savard); University of California Los Angeles David Geffen School of Medicine, Los Angeles, California, USA (R.M. Humphries)

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Table 2

Sequences of primers and probes used for identification of NDM–producing isolates, United States, April 2009–March 2011*

Assay Primers and probes Sequence, 5′ → 3′
Real-time PCR:NDM/KPC screen
NDM, forward primer GAC CGC CCA GAT CCT CAA
NDM, Reverse primer CGC GAC CGG CAG GTT
NDM, probe (HEX)† TG GAT CAA GCA GGA GAT
KPC, forward primer GGC CGC CGT GCA ATA C
KPC, reverse primer GCC GCC CAA CTC CTT CA
KPC, probe (FAM)† TG ATA ACG CCG CCG CCA ATT TGT
16S, forward primer TGG AGC ATG TGG TTT AAT TCG A
16S, reverse primer TGC GGG ACT TAA CCC AAC A
16S, probe (CY5)†
CA CGA GCT GAC GAC AR‡C CAT GCA
DNA sequence analysis§
NDM-1 forward ACT CGT CGC AAA GCC CAG
NDM-1 reverse
CTC ATG TTT GAA TTC GCC C
Internal DNA sequencing primers
NDM-2F ACA AGA TGG GCG GTA TGG A
NDM-2R
CGT CCA TAC CGC CCA TCT
DIG-labeled probe synthesis NDM-F1 GAA TTG CCC AAT ATT ATG CAC C
NDM-R1 AGC GCA GCT TGT CGG CCA TG

*NDM, New Delhi metallo-β-lactamase; KPC, Klebsiella pneumoniae carbapenemase; DIG, digoxigenin.
†NDM probes were labeled with HEX, KPC probes were labeled with FAM, and 16S probes were labeled with CY5 at their 5′ ends. Each contained a black hole quencher at the 3′ end.
‡R, A or G (International Union of Biochemistry codes for DNA bases).
§Amplification using primers NDM-1 forward and NDM-1 reverse, both located outside the coding region of blaNDM, results in a 1,013-bp product.

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