Development Of Enzyme-Linked Dna-Based Electrochemical Biosensor For Rapid Detection Of Acinetobacter baumannii

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dc.contributor.authorKanapathy, Swarnaletchumi
dc.date.accessioned2017-09-07T07:23:45Z
dc.date.available2017-09-07T07:23:45Z
dc.date.issued2015-02
dc.description.abstractAcinetobacter baumannii has been recognized as an emerging nosocomial pathogen and well known as a multi-drug resistant bacterium. It has also been identified as an important cause of morbidity and mortality in hospitals, especially among critically ill patients. Early diagnosis of infection caused by A. baumannii is a major strategy in controlling the infection caused by this pathogen. Current identification of this bacterium is by conventional culture method and biochemical tests, which takes about 2 to 7 days to produce results. These routine procedures are time consuming and labour intensive. Hence, there is a need for a new rapid, sensitive, specific and economical test that would allow rapid management of A. baumannii infections. Currently, polymerase chain reaction (PCR) has been widely used in diagnostic field for the detection of blaOXA-51-like gene in A. baumannii. Nevertheless, the detection of amplified product using agarose gel analysis has its own drawback such as time consuming and exposure to hazardous element like ethidium bromide and UV light. Therefore, the objective of this study is to develop an enzyme-based electrochemical genosensor for asymmetric PCR (aPCR) amplicons detection of blaOXA-51-like gene in A. baumannii. The biotin labelled DNA probe was immobilized on streptavidin modified screen-printed carbon electrode (SPCE) and the complementary ssDNA target with fluorescein labelled was hybridize to the probe. The amperometric current response was detected with peroxidase conjugated anti-fluorescein antibody. The limit of detection of the developed assay using synthetic oligomer was found to be 0.008 μM. The analytical sensitivity of the assay at genomic level and bacterial cell level was 0.5 pg and 103 CFU/ml, respectively. The diagnostic performance of the assay showed to be 100% specific and sensitive in detection of A. baumannii. Based on accelerated stability performance, this enzyme-based genosensor was estimated to be stable for approximately 1.2 years when stored at 4 °C. In conclusion, the results of this study showed that the blaOXA-51-like gene is specific for A. baumannii and the developed electrochemical genosensor will be useful in providing rapid treatment and management to the infected patients.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/4524
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectDevelopment enzyme-linked DNAen_US
dc.subjectfor rapid detection of acinetobacter baumanniien_US
dc.titleDevelopment Of Enzyme-Linked Dna-Based Electrochemical Biosensor For Rapid Detection Of Acinetobacter baumanniien_US
dc.typeThesisen_US
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