alexa PCR-based Method for Rapid and Minimized Electrochemical Detection of mecA Gene of Methicillin-resistant Staphylococcus aureus and Methicillinresistant Staphylococcus epidermidis
ISSN: 2327-5146

General Medicine: Open Access
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Research Article

PCR-based Method for Rapid and Minimized Electrochemical Detection of mecA Gene of Methicillin-resistant Staphylococcus aureus and Methicillinresistant Staphylococcus epidermidis

Tomohiko Ikeuchi 1, Masafumi Seki 2,3*, Yukihiro Akeda4, Norihisa Yamamoto2, Shigeto Hamaguchi2, Tomoya Hirose5, Keiichiro Yamanaka1,Masato Saito1, Kazunori Tomono1 and Eiichi Tamiya1
1Department of Applied Physics, Graduate School of Engineering, Osaka, Japan
2Division of Infection Control and Prevention, Osaka University Hospital, Osaka, Japan
3Division of Respiratory Medicine and Infection Control, Tohoku Pharmaceutical University Hospital, Sendai, Japan
4Laboratory of Clinical Research on Infectious Diseases, Research Institute for Microbial Diseases, Osaka, Japan
5Department of Traumatology and Acute Critical Medicine, Osaka University Hospital, Osaka University, Suita City, Osaka, Japan
Corresponding Author : Masafumi Seki
M.D., Ph.D., Division of Respiratory
Medicine and Infection Control, Tohoku Pharmaceutical University Hospital
1-12-1 Fukumuro, Miyagino, Sendai, 983-8512, Japan
Tel: +81-22-259-1221
Fax: +81-22- 259-0507
E-mail: [email protected]
Received November 14, 2015; Accepted November 30, 2015; Published December 07, 2015
Citation: Ikeuchi T, Seki M, Akeda Y, Yamamoto N, Hamaguchi S, et al. (2015) PCR-based Method for Rapid and Minimized Electrochemical Detection of mecA Gene of Methicillin-resistant Staphylococcus aureus and Methicillin-resistant Staphylococcus epidermidis. Gen Med (Los Angel) 3: 215. doi:10.4172/2327-5146.1000215
Copyright: © 2015 Ikeuchi T, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important pathogens that cause nosocomial infections. However, microbiological culture techniques take a few days to yield results; therefore, a simple, cost-effective, and rapid detection system is required for screening for MRSA and related bacteria: Methicillinresistant Staphylococcus epidermidis (MRSE) carriers during the hospital admissions process. In this study, we described the simplified method using by one-time use and screen-printed carbon electrodes, relied upon current quantification of Hoechst dyes which bound with DNA amplified via polymerase chain reaction (PCR) targeted for MRSA mecA gene. Amount of DNA-bound Hoechst molecules were measured by the hand-held potentiostat within two minutes. We found that the peak of a Hoechst-mediated current depended upon the number of MRSA cells, and successfully distinguished between carriers and a non-carrier based on nasal swabs from the patients. This method required only 10 μL for application, and the results could be obtained within total 60 min from sample collection when a minimum of 1 × 103 MRSA cells was present. These results suggested that this minimized technique has the potential to become a useful system of active surveillance for MRSA/MRSE carriers.

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