Magnetic nanoparticle based PCR assay for detection of Campylobacter species in food samples and dielectrophoretic impedance measurement for DNA detection
5
Issued Date
2023
Copyright Date
2014
Language
eng
File Type
application/pdf
No. of Pages/File Size
xiii, 82 leaves : ill.
Access Rights
restricted access
Rights Holder(s)
Mahidol University
Bibliographic Citation
Thesis (Ph.D. (Medical Technology))--Mahidol University, 2014
Suggested Citation
Wuttichote Jansaento Magnetic nanoparticle based PCR assay for detection of Campylobacter species in food samples and dielectrophoretic impedance measurement for DNA detection. Thesis (Ph.D. (Medical Technology))--Mahidol University, 2014. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/89502
Title
Magnetic nanoparticle based PCR assay for detection of Campylobacter species in food samples and dielectrophoretic impedance measurement for DNA detection
Alternative Title(s)
การพัฒนาวิธี PCR โดยใช้ magnetic nanoparticle เพื่อตรวจหา Campylobacter species ในตัวอย่างอาหารและการใช้เทคนิค dielectrophoretic impedance ในการตรวจวัดดีเอ็นเอ
Author(s)
Abstract
Campylobacter species are major foodborne pathogens, causing bacterial gastroenteritis worldwide. Poultry are considered to be important reservoirs of Campylobacter transmission and campylobacteriosis commonly relates to the consumption of undercooked chicken products. This study successfully developed a magneto-PCR-enzyme linked gene assay for the detection of thermophilic Campylobacter from chicken skin. The forward primers immobilized on magnetic nanoparticles captured Campylobacter DNA to be amplified by PCR reaction and the PCR products were detected by streptavidin-horseradish peroxidase and its substrate. The method demonstrated 100% specificity and high sensitivity with one picogram detection of Campylobacter DNA. All positive results derived from the detection in naturally contaminated samples were in accordance with the results analyzed by a conventional cultural method and PCR assay. In this study, the dielectrophoretic impedance measurement (DEPIM) method has been used for the detection of PCR and LAMP products by measuring an increment of the conductance. Dielectrophoresis is a motion of the polarized particles in spatially non-uniform electric fields. The DEP- trapping process of DNA, generated from PCR or LAMP products, was not visualized by a fluorescence microscopy. However, in an AC electric field of 150 kHz, the conductance increased when applying the undiluted PCR product of 187 base pairs. After optimizing conditions for detecting LAMP product, only undiluted and ten-fold diluted LAMP products significantly demonstrated the conductance increment. This preliminary study showed the possibility of applying The DEPIM method for detection of PCR and LAMP products.
Degree Name
Doctor of Philosophy
Degree Level
Doctoral Degree
Degree Department
Faculty of Medical Technology
Degree Discipline
Medical Technology
Degree Grantor(s)
Mahidol University