Development of BKV-quantum dot nanoparticles (QDS) based on lateral flow chromatographic test strip
1
Issued Date
2024
Copyright Date
2019
Resource Type
Language
eng
File Type
application/pdf
No. of Pages/File Size
xvi, 108 leaves : ill.
Access Rights
open access
Rights
ผลงานนี้เป็นลิขสิทธิ์ของมหาวิทยาลัยมหิดล ขอสงวนไว้สำหรับเพื่อการศึกษาเท่านั้น ต้องอ้างอิงแหล่งที่มา ห้ามดัดแปลงเนื้อหา และห้ามนำไปใช้เพื่อการค้า
Rights Holder(s)
Mahidol University
Bibliographic Citation
Thesis (M.Sc. (Materials Science and Engineering))--Mahidol University, 2019
Suggested Citation
Tanapon Sioloetwong Development of BKV-quantum dot nanoparticles (QDS) based on lateral flow chromatographic test strip. Thesis (M.Sc. (Materials Science and Engineering))--Mahidol University, 2019. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/92179
Title
Development of BKV-quantum dot nanoparticles (QDS) based on lateral flow chromatographic test strip
Author(s)
Abstract
BK virus (BKV) is a virus in the polyomavirus family that have a characteristic structure such as an icosahedral shape and a non-envelop capsid structure. This virus can develop into inflammatory diseases such as pneumonia, nephritis, etc. In clinical manifestation, BKV associated nephropathy (BKVAN) was a mostly founded patient which affect the kidney transplantation from donor to recipient. Because BKVAN can cause allograft dysfunction and allograft failure. Nowadays, the screening test of BKVAN was required more quantity of viruses, more time, and more steps. Therefore, lateral flow technology has been developed to point-of-care for reduced detection time and cost. Lateral flow assay (LFAs) is widely used in the biosensor technique due to easily used, rapidly test, low cost, high sensitivity, high specificity, etc. However, LFAs has a limited to detect in qualitative analysis. Usually, gold nanoparticles (AuNPs) is used as the conventional labeling particles in LFAs, because AuNPs have excellent stability and excellent optical property. In contrast, AuNPs cannot readout signals in quantitative analysis. Therefore, the alternative particle is considerable. Quantum Dots (QDs) is the alternative particle that has unique optical properties such as strong photostability, high quantum yield fluorescence, and tunable emission wavelength. Consequently, QDs is a choice for applied in quantitative analysis. In this work, QDs-DNA was used as a labeling reporter for detecting LAMP-BKV products based on lateral flow assay. The results divided into two parts: (1) synthesis of QDs-DNA and (2) applying QDs-DNA to detect LAMP-BKV based on LFAs. For the synthesis of labeling reporter, the optical properties showed the different maximum emission wavelengths of CdSe QDs (lambda(em)=590 nm), CdSe/CdZnS CSQDs (lambda(em)=637 nm), and QDs -COOH (lambda(em)= 629 nm). From the emission results, CSQDs and QDs-COOH affected the quantum confinement effect when compared with core CdSe QDs. The size calculation of CdSe QDs and CSQDs was 3.4 nm and 11.7 nm, respectively. Also, CSQDs showed a high quantum yield efficiency at 73%. The miscibility of QDs-COOH showed the hydrophilic property after surface treatment to the COOH group. Then, FTIR spectra showed a relative wavenumber with the carboxylic group, such as hydroxyl group ((approximately equal to) 3,300 cm-1), carbonyl group ((approximately equal to) 1650 cm-1), and coordination of Cd2+ and S2- of carboxylic precursor ((approximately equal to) 500cm-1). The zeta potential of QDs-COOH was optimized into -30 mV for improving the particle stability by DLS measurement. QDs-DNA presented a characteristic peak at 260 nm which referred to the electron delocalization of nucleosides. The reduction of the fluorescent intensity of QDs-DNA confirmed the DNA immobilization because of losing QDs during DNA immobilization and transferring electrons from QDs to DNA. Then, QDs-DNA exhibited the most massive weight compared with QDs-COOH and DNA probe by gel electrophoresis. For optimization of QDs-DNA and BKV LAMP product based on lateral flow assay, the suitable of particle flowability and particle absorbability were 200 µL of CMF buffer with Tween 20. The optimum interfering reaction was 50 µL of 100 nM of QDs-DNA. Then, the optimum concentration of streptavidin loading was 10-5 M of both test line and control line. The sensitivity of LFAs was observed by adjusting the volume ratios of QDs-DNA and LAMP-BKV. The result showed that signals were disappeared on both lines, although the volume ratios of the LAMP-BKV product would be changed. In summary, the improvement solution of QDs based on LFAs detection with LAMP-BKV product will suggest promoting in 4 priorities: (1) The signal intensity on test line will be evaluated the concentration of streptavidin for increasing the signal of detection, because many works of literature applied the high concentration of streptavidin to detect by dispenser machine which can reduce the interfering reaction with high concentration of protein loading; (2) The DNA hybridization should apply the hybridization temperature and time to raise signal on control line; (3) The alignment of LFAs testing and membrane types will be changed for applying temperature on ground test and improving the hybridization time, respectively: (4) The manufacturing of protein lining will apply to use a dispenser machine due to controlling the protein bandwidth for detection. In summary, the improvement of QDs-LFAs can dissolve the limitation of detection which include bio-recognition interaction, lining of protein alignment and reproducibility by dispenser manufacturing
Description
Materials Science and Engineering (Mahidol University 2019)
Degree Name
Master of Science
Degree Level
Master's degree
Degree Department
Faculty of Science
Degree Discipline
Materials Science and Engineering
Degree Grantor(s)
Mahidol University