MSc in Biotechnology
Dissertation Project – 2nd Cycle
Aprovado pela CC do MBiotec em 20-05-2015
Student Name: Cindy Duarte de Oliveira
Student email address: cd.oliveira@campus.fct.unl.pt
No.44697
Supervisor(s): Professor Pedro Viana Baptista
Supervisor(s) email address: pmvb@fct.unl.pt
Lab/Institution: CIGMH, Departamento de Ciências da Vida, FCT, UNL
Title: Gold Nanoparticles: Development of a rapid molecular test for detection of pathogenic agents in
Food Industry
BACKGROUND
Each year, millions of people get sick from contaminated food (poultry, beef, pork, eggs, milk, seafood,
nut products, and fresh produce). Symptoms of food poisoning include upset stomach, abdominal
cramps, nausea and vomiting, diarrhea, fever, and dehydration. These are caused by pathogenic agents
such as Salmonella, Staphylococcus aureus and Clostridium. The traditional tests which are used to
detect these pathogens are expensive, time-consuming and labor-intensive. Thus, it is necessary the
development of rapid and sensitive detection methods for the food industry in order to control the
There
tjh and prevent the outbreaks of these pathogens. Recently, a novel nucleic acid amplification
spread
assay called loop-mediated isothermal amplification (LAMP) has been adapted to this type of detection
for being rapid, specific and simpler than others. In the last decade gold nanoparticles have also been
used in molecular tests due to its unique properties. Metallic nanoparticles have high values of
dispersion and absorption in UV/ visible range which allow them to have high optic sensibility in optical
detection methods. Thus, they are a great choice in colorimetric detection tests.
OBJECTIVES
This Project aims to develop a rapid, sensitive and low cost molecular test for detection of pathogenic
agents in Food Industry. Based on gold nanoparticles and their colorimetric properties, they will be
functionalized with thiol-modified DNA oligonucleotides (Gold nanoprobes), allowing the detection of
specific DNA targets amplified by LAMP.
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MSc in Biotechnology
Dissertation Project – 2nd Cycle
PROJECT DESCRIPTION
The Project will involve:
I)
In silico design of probes with the ideal characteristics (sequence, hybridization) to detect
the intended targets;
II)
Synthesis and functionalization of AuNPs with thiolated oligonucleotides;
III)
Characterization of Au-nanoprobes for non-cross-linking based DNA detection;
IV)
LAMP design and optimization for DNA retrieved from potential pathogenic agents (S.
aureus, Salmonella sp….);
V)
LAMP coupled with Au-nanoprobe performance assessment;
VI)
Validation against PCR gold-standard (collaboration with SME);
VII)
Thesis writing.
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