Nucleic Acid Amplification Free-QCM-DNA Biosensor for Burkholderia pseudomallei Detection
Issued Date
2023-12-01
Resource Type
ISSN
03438651
eISSN
14320991
Scopus ID
2-s2.0-85174518623
Journal Title
Current Microbiology
Volume
80
Issue
12
Rights Holder(s)
SCOPUS
Bibliographic Citation
Current Microbiology Vol.80 No.12 (2023)
Suggested Citation
Suvanasuthi R., Cheewasatheinchaiyaporn T., Wat-aksorn K., Promptmas C. Nucleic Acid Amplification Free-QCM-DNA Biosensor for Burkholderia pseudomallei Detection. Current Microbiology Vol.80 No.12 (2023). doi:10.1007/s00284-023-03490-y Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/90823
Title
Nucleic Acid Amplification Free-QCM-DNA Biosensor for Burkholderia pseudomallei Detection
Author's Affiliation
Other Contributor(s)
Abstract
Burkholderia pseudomallei is a gram-negative bacterium that causes the infectious disease melioidosis, a disease that can still be fatal despite appropriate treatment. The bacterium contains the gene clusters for the type III secretion system (TTSS), which are essential for its pathogenicity. This gene was often employed for accurate diagnosis through the laborious process of gene amplification. This work intends to develop a quartz crystal microbalance (QCM)-based TTSS gene detection method without gene amplification approaches to simplify the diagnosis process. In this study, it was demonstrated that a 540 bp sequence flanked by BglI restriction sites within the TTSS1 on the B. pseudomallei genome is an effective target for specific detection of the bacteria. After cultivation and genome extraction, the bacteria can be detected by digesting its genome with BglI in which the TTSS1 fragment is detected by a QCM-DNA biosensor, eliminating the need for nucleic acid amplification. A specific probe designed to bind to the TTSSI fragment was utilized as the receptor on the QCM-DNA biosensor which provided the ability to detect the fragment. The limit of detection of the QCM-DNA biosensor was 0.4 µM of the synthetic DNA target oligonucleotide. The system was also capable of specifically detecting the BglI digested-DNA fragment of B. pseudomallei species with significantly higher signal than B. thailandensis. This study provides evidence for an effective QCM-DNA biosensor that can identify B. pseudomallei without the need for nucleic acid amplification.