PCR-Based Method for Rapid and Minimized Electrochemical Detection of mecA Gene of Methicillin-Resistant Staphylococcus aureus and Methicillin-Resistant Staphylococcus epidermis
Journal Title: Global Journal of Infectious Diseases and Clinical Research - Year 2016, Vol 2, Issue 1
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: Methicillin-resistant 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 isolates, 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 isolates was present. These results suggested that this minimized technique has the potential to become a useful system of active surveillance for MRSA/MRSE carriers.
Authors and Affiliations
Ikeuchi Tomohiko, Seki Masafumi, Akeda Yukihiro, Yamamoto Norihisa, Hamaguchi Shigeto, Hirose Tomoya, Yamanaka Keiichiro, Saito Masato, Tomono Kazunori, Tamiya Eiichi
Keywords
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- EP ID EP334758
- DOI 10.17352/2455-5363.000007
- Views 102
- Downloads 0
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