Detection and quantification of COMP neoepitope in equine saliva: A biomarker for detection of early stages of Osteoarthritis
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Typ
Examensarbete för masterexamen
Program
Biotechnology (MPBIO), MSc
Publicerad
2021
Författare
Lord, Moa
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
Osteoarthritis (OA) is a chronic in
ammatory joint disease, with painful destructive processes
locally in the joint. It is not possible to diagnose the early destructive biochemical
events within the joint that leads to pain and subsequent lameness. Many horses with
local joint pain due to early OA with a low-grade in
ammation are used today in the
equestrian industry. This disease develops over time and is usually not diagnosed until the
end stages, where irreversible damage and clinical symptoms can be noticed. A biomarker
for early matrix degradation of articular cartilage is needed as a diagnostic tool for early
diagnosis of OA. A speci c neoepitope of cartilage oligomeric matrix protein (COMP) have
previously been identi ed and shown to increase in synovial
uid and serum from horses
with acute lameness. The aim of this thesis was to detect and quantify the speci c COMP
neoepitope in saliva sampled from healthy horses and horses with OA. This aim included
evaluation of sample collection and preparation, sample collection throughout the day and
validation of the capillary western blot Wes™by inhibition ELISA. Salivette®, a speci c
collection device developed for salivary collection, was used to collect the saliva. Saliva
sampled on the horse tongue, no food intake within one hour, obtained lower background
noise, showed higher chemiluminescence signal and more de ned peaks compared to saliva
sampled close to food intake, when analysed with Wes™. In Wes™, the COMP neoepitope
could be detected at the apparent molecular weight of 58 kDa. The concentration of the
COMP neoepitope was determined with inhibition ELISA developed for quanti cation of
COMP neoepitope in synovial
uid and serum from horses. The highest concentration
of COMP neoepitope was quanti ed in a horse recently diagnosed with OA, compared to
saliva from healthy horses. This indicates that saliva from horses can be used for detection
and quanti cation of COMP neoepitope. However, this thesis included a limited amount
of horses and they were not clinically examined by a veterinarian prior to saliva sampling.
A larger group of well-de ned healthy horses and horses with OA is required to draw
statistical conclusions. Quanti cation of COMP neoepitope in saliva is a step toward the
future goal of developing a point of care diagnostics, which can be an accessible tool for
diagnosis of OA, monitoring the horse during rehabilitation and also be used in everyday
training of athletic horses.
Beskrivning
Ämne/nyckelord
Osteoarthritis , Biomarker , COMP neoepitope , Saliva , Wes™ , Inhibition ELISA