Urinary tract infection diagnostic
A urinary tract infection, horizontal technology, applied in the direction of disease diagnosis, medical automated diagnosis, instruments, etc.
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Embodiment 1
[0281] Example 1 - Selection of biomarkers
[0282] Samples were collected as part of an I4I-funded project in which a total of 189 urine samples from adult women presenting with suspected uncomplicated UTI were comprehensively analyzed for 60 biomarkers.
[0283] There are two goals:
[0284] i. Define a panel of biomarkers to differentiate between negative and positive cultures;
[0285] ii. Select the biomarker that provides the best discrimination between suspected UTI samples and samples from recovered patients (healthy).
[0286] To differentiate between patients presenting with different types of infection, binary logistic regression was used to predict which biomarkers were significant for the outcome on their own without interaction with other parameters. Significant markers in this analysis were then further analyzed using R language Stuttgart Neural Network Simulation (RSNNS) (Stuttgart Neural Network Simulation in R), where self-significant biomarkers were weight...
Embodiment 2
[0303] Example 2 - Development of a Lateral Flow Assay
[0304] For lateral flow development, three selected biomarkers, HNE, MMP8 and cystatin C, were employed. Using a standard lateral flow format, three lateral flow assays were developed and optimized, packaged into single-well plastic housings supplied by Bibby sterylin.
[0305] HNE lateral flow
[0306] Preparation of materials: Capture line (anti-HNE BSA-PEG Fab (Alere San Diego, cat. no. 01241)) and control line (BSA-biotin) were immobilized at 1 mg / ml in PBS+1% sucrose using an Imagene Isoflow dispenser on a nitrocellulose membrane (Sartorius, CN140). Membranes were then dried in a tunnel dryer (Hedinair) at 60°C and stored with desiccant until use.
[0307] Gold conjugates were prepared according to known experimental procedures. Anti-HNE BSA-PEG Fab (Alere San Diego, Cat. No. 01871) was conjugated to 40 nm gold colloids to a final concentration of 15 μg / ml in suspension buffer of 20 mM MES, pH 5.3. After 10 m...
Embodiment 3
[0325] Example 3 - Statistical Analysis and Algorithm Development
[0326] In the Cardiff POETIC study (Bates 2014), a total of 424 samples were collected: 204 were Visit 1 samples and 220 were Visit 2 samples (not all of these were matched samples). In total, there are 93 matching Visit 1 and Visit 2 samples.
[0327] Sample information is available for a total of 194 samples: 175 samples are Visit 1 samples and 19 samples are Visit 2 samples. Of the 93 matched samples, 81 Visit 1 samples had incidental clinical information.
[0328] Of the 81 Visit 1 samples, 32 samples were UTI positive and 49 samples were UTI negative. Status based on microbiological results:
[0329] No growth = no growth found
[0330] · No significant growth = growth of pure isolate 5
[0331] · Mixed 2 kinds of microorganisms (orgs) = 2 kinds of microorganisms in 10 5 cfu / mL or more growth
[0332] Mixed > 2 microorganisms = 3 or more isolates with 10 5 cfu / mL or more growth
[0333] · POS = p...
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