PCR-Dipstick Chromatography for Differential Detection of Carbapenemase Genes Directly in Stool Specimens

Rathina Kumar Shanmugakani; Yukihiro Akeda; Norihisa Yamamoto; Noriko Sakamoto; Hideharu Hagiya; Hisao Yoshida; Dan Takeuchi; Yo Sugawara; Takuya Kodera; Mitsuo Kawase; Warawut Laolerd; Narong Chaihongsa; Pitak Santanirand; Yoshikazu Ishii; Shigeyuki Hamada; Kazunori Tomono

doi:10.1128/AAC.00067-17

PCR-Dipstick Chromatography for Differential Detection of Carbapenemase Genes Directly in Stool Specimens

Antimicrob Agents Chemother. 2017 May 24;61(6):e00067-17. doi: 10.1128/AAC.00067-17. Print 2017 Jun.

Authors

Rathina Kumar Shanmugakani^{1

2

3}, Yukihiro Akeda^{4

2

3}, Norihisa Yamamoto^{1

2

3}, Noriko Sakamoto³, Hideharu Hagiya^{1

2

3}, Hisao Yoshida^{1

2

3}, Dan Takeuchi³, Yo Sugawara³, Takuya Kodera⁵, Mitsuo Kawase^{5

6}, Warawut Laolerd⁷, Narong Chaihongsa⁷, Pitak Santanirand⁷, Yoshikazu Ishii⁸, Shigeyuki Hamada³, Kazunori Tomono^{1

2}

Affiliations

¹ Department of Infection Control and Prevention, Graduate School of Medicine, Osaka University, Osaka, Japan.
² Division of Infection Control and Prevention, Osaka University Hospital, Osaka, Japan.
³ Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
⁴ Department of Infection Control and Prevention, Graduate School of Medicine, Osaka University, Osaka, Japan [email protected].
⁵ Tohoku Bio-Array, Sendai, Japan.
⁶ Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan.
⁷ Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
⁸ Department of Microbiology and Infectious Diseases, Toho University School of Medicine, Tokyo, Japan.

Abstract

A PCR-dipstick chromatography technique was designed and evaluated for differential identification of bla_NDM, bla_KPC, bla_IMP, and bla_OXA-48 carbapenemase genes directly in stool specimens within 2 h. It is a DNA-DNA hybridization-based detection system where PCR products can be easily interpreted by visual observation without electrophoresis. The PCR-dipstick showed high sensitivity (93.3%) and specificity (99.1%) in directly detecting carbapenemase genes in stool specimens compared with multiplex PCR for genomic DNA of the isolates from those stool specimens.

Keywords: Enterobacteriaceae; carbapenemase; molecular diagnostics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics*
Carbapenems / pharmacology
Feces / microbiology*
Humans
Molecular Diagnostic Techniques / methods*
Multiplex Polymerase Chain Reaction / methods*
beta-Lactamases / genetics*

Substances

Bacterial Proteins
Carbapenems
beta-Lactamases
carbapenemase