Ann Lab Med.  2012 Sep;32(5):359-361. 10.3343/alm.2012.32.5.359.

Multiplex PCR for Rapid Detection of Genes Encoding Class A Carbapenemases

• Hong SS
• Kim K
• Huh JY
• Jung B
• Kang MS
• Hong SG

• KMID: 1387351
• DOI: http://doi.org/10.3343/alm.2012.32.5.359

Abstract

In recent years, there have been increasing reports of KPC-producing Klebsiella pneumoniae in Korea. The modified Hodge test can be used as a phenotypic screening test for class A carbapenamase (CAC)-producing clinical isolates; however, it does not distinguish between carbapenemase types. The confirmation of type of CAC is important to ensure optimal therapy and to prevent transmission. This study applied a novel multiplex PCR assay to detect and differentiate CAC genes in a single reaction. Four primer pairs were designed to amplify fragments encoding 4 CAC families (SME, IMI/NMC-A, KPC, and GES). The multiplex PCR detected all genes tested for 4 CAC families that could be differentiated by fragment size according to gene type. This multiplex PCR offers a simple and useful approach for detecting and distinguishing CAC genes in carbapenem-resistant strains that are metallo-beta-lactamase nonproducers.

Keyword

Carbapenemase; Multiplex PCR; KPC; GES

MeSH Terms

Bacterial Proteins/*genetics/metabolism
DNA Primers/metabolism
Databases, Genetic
Humans
Klebsiella Infections/microbiology
Klebsiella pneumoniae/genetics/isolation & purification/metabolism
*Multiplex Polymerase Chain Reaction
beta-Lactamases/*genetics/metabolism

Figure

• 

  Fig. 1 Results of multiplex PCR for class A carbapenemase (CAC)-producing strains. Multiplex PCR products were separated on a 2% agarose gel. Lanes 1 and 14 show the 100-bp DNA ladder; lane 2, the PCR product of the negative control (distilled water); lanes 3 and 4, KPC-type enzyme-producing strains; lane 5, SME-type; lanes 6 and 7, NMC-A and IMI-type, respectively; lanes 8-13, GES-type. The amplified product from each PCR is indicated on the right, and the size of the marker in base pairs is shown on the left.

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