Investigative Techniques Commons^™

A Review Of The Pathogenesis, Clinical Features And Diagnostic Indicators Of The Novel Condition Vaccine-Induced Thrombotic Thrombocytopenia, Ada Fleming

International Undergraduate Journal of Health Sciences

The introduction of vaccination against SARS-CoV-2 infection was a welcome and significant event in the COVID-19 pandemic. While vaccine administration was for the most part successful, it did come with the emergence of a novel condition, Vaccine-Induced Thrombotic Thrombocytopenia (VITT). This condition presents after the administration of adenoviral vector-based vaccines against COVID-19, causing thrombocytopenia and thrombosis in affected individuals. There have been 10 suspected cases of VITT in Ireland. While its mechanism is not fully understood, the condition is characterised by the presence of Platelet Factor 4 (PF4) antibodies. There are several laboratory indicators which may suggest that a patient …

Changing Diagnostic Methods And Increased Detection Of Verotoxigenic Escherichia Coli, Ireland, Thomas Rice, Noreen Quinn, Roy D. Sleator, Brigid Lucey

Department of Biological Sciences Publications

The recent paradigm shift in infectious disease diagnosis from culture-based to molecular-based approaches is exemplified in the findings of a national study assessing the detection of verotoxigenic Escherichia coli infections in Ireland. The methodologic changes have been accompanied by a dramatic increase in detections of non-O157 verotoxigenic E. coli serotypes.

The Analysis And Characterisation Of Rotavirus And Norovirus Circulating In The South Of Ireland, Norma Reidy

Theses

On average more than 3,000 cases of rotavirus diarrhea are detected annually amongst Irish children less than 3 years of age. Rotavirus disease occurs mainly during the winter months, with peak infections detected from February to April. The symptoms of disease are diarrhea and dehydration, with 80% of patients requiring hospitalisation, The mean duration of hospitalisation is 3-4 days, with an average cost per day of €409. Eighteen percent of infections were detected in infants less than 6 months of age, suggesting inadequate maternal derived protection from existing strains, or the existence of novel strains in Ireland with little or …

Validation Of An Immunomagnetic-Electrochemiluminescence Detection System For The Detection Of Escherichia Coli 0157 In Food And Comparison With An Iso Standard Method, Eimear Carney

Theses

Rapid and sensitive methods for detecting Escherichia coli 0157 in contaminated food, water and other biological samples are important in limiting and preventing the spread of this pathogen. Traditional culture based methods for bacterial identification and detection can be time consuming and labour intensive. Modem alternative methods should be faster whilst maintaining sensitivity. Immunomagnetic separation (IMS) methodology has been investigated and appears promising for rapid bacterial detection in unrelated food and environmental samples. In this study, a commercial sensor which combined IMS with electrochemiluminescence (ECL) was evaluated for the detection of E. coli 0157 in unrelated food matrices. Emitted signals …

Applications Of The Atomic Force Microscope Within The Fields Of Haematology And Cytogenetics, Margaret O'Reilly-Crowley

Theses

Atomic force microscopy [AFM] has become a rapidly developing interdisciplinary field of research in recent years. The technique, including instrumentation and specimen preparation methods, is now more sophisticated and its application has increased dramatically. The key advantage of AFM is in providing three-dimensional images of samples imaged. Both size and shape of samples imaged can be obtained allowing their subsequent quantification. Red blood cells [RBCs] and chromosomes were imaged in this work. A brief overview of the current status of biomedical AFM, has been presented in the review.

Measurement of mean cell volume [MCV] of immobilised red blood cells with …

The Molecular Epidemiology Of Rotavirus In Ireland, Fiona O'Halloran

Theses

Between 1997 and 1998, 3,136 cases of rotavirus diarrhoea were detected in Irish children less than 2 years of age. Hospital inpatients accounted for 80% of these infections, with the remainder being diagnosed in general practice. A large percentage of infections were detected in neonates, suggesting a possible inadequate maternal derived protection from existing indigenous strains. This feature suggested the possible existence of ‘novel’ strains circulating in Ireland. No data describing the epidemiology of rotavirus strains in this country currently exists. Furthermore no assessment of the potential health-economic impact or quantitation of potential disease burden was ever undertaken

Three hundred …

The Atomic Force Microscope In The Elucidation Of Leukocyte Abnormality, Derbrenn O'Connor

Theses

The aim of this project was to investigate the novel use of the atomic force microscope (AFM) as a technique in the elucidation of leukocyte abnormality. To this end, comparative AFM imaging and elasticity studies were performed on normal and myeloid leukaemic leukocytes.

Initially, an evaluation of techniques for the isolation of specific leukocyte (e.g. lymphocytes, granulocytes, etc.) populations from whole blood or leukocyte concentrates, was carried out. It was found that a silanised glass surface failed to select and immobilise a leukocyte population. However, centrifugation of whole blood in an Isopaque-ficoll gradient solution yielded a viable mixed leukocyte population …

Evaluation Of The Role Of Fructosamine As An Indicator Of Diabetic Control, Katherine Hooley

Theses

Diabetes mellitus, a common metabolic disease, is characterised by excess circulating glucose concentrations. Such a hyperglycaemic status provides the stimulus for irreversible glycation (non-enzymatically post-translational) of structural and functional proteins. Protein glycation, although a cause of complication (neuropathy, micro/macrovascular disease), provides a time-averaged index of diabetes status and control. In particular, glycated plasma proteins analysis, i.e., the fructosamine assay, has received favourable response due to its speed, simplicity, low cost, and ease of automation. However, lack of standardisation and a universally-accepted calibrant has limited technical and subsequent clinical confidence in fructosamine analysis. In response, a comprehensive technical evaluation (including novel …

Development Of A Differential Assay To Selectively Measure Α-Amylase Isoenzymes., Miriam O'Connor

Theses

α-Amylase is a heterogeneous polypeptide, primarily found in the Salivary and Pancreatic glands, giving rise to two isoenzymes, namely Salivary- and Pancreatic-type Amylases. Total α-Amylase activity measurements are routinely carried out in hospital laboratories to aid in the diagnosis of diseases of the pancreas and in the investigation of pancreatic function in patients with severe abdominal pains. With the development and wide availability of modem chromogenic oligosaccharide substrates specific for Amylase, assays have become more sensitive, rapid and easier to use.

Differential Amylase Isoenzyme assays ware developed to distinguish isoenzyme levels. Several methods were developed for isoenzyme determination. Of these. …

Development Of An Elisa Method For The Measurement Of Cd44 Expression In Human Serum, Ciara Hurley

Theses

CD44 is a cell surface glycoprotein (mol. wt. =85-95 kDa) which has been implicated in lymphocyte-HEV interactions and the process of lymphocyte homing. CD44 is a complex glycoprotein which undergoes extensive alternative splicing to generate various CD44 isoforms. CD44 functions include the delivery of an activation signal to T-cells, binding to hyaluronate, mediation of lymphocyte homing, and the spread of metastatic cells. CD44 has been particularly implicated in the dissemination of non- Hodgkin's lymphoma and as such may serve as a useful prognostic marker of disease progression. CD44 measurement in serum may provide a rapid, inexpensive and relatively non-invasive prognostic …

Development And Validation Of An Improved Ultrasensitive Assay For Cardiac Troponin I (Ctni), William G. Blake

Theses

Cardiac troponin I (cTnl) has been reported to be a highly specific marker for myocardial injury. In patients suffering from acute coronary syndromes, a sensitive serum marker which is closely related to the extent of myocardial damage which provides prognostic data and which can be measured rapidly is desirable. In this thesis a “sandwich” enzyme immunoassay was developed to measure cTnI. The developed assay was reproducible with the inter assay precision ranging from 4.8-14% and the intra assay precision ranging from 3.1-9.5%. The limit of detection of the assay was calculated as 0.385 ng/ml. The upper reference range (non-parametric 95% …

Methicillin Resistant Staphylococcus Aureus (Mrsa): Molecular Detection And Dna Fingerprinting, Leslie Cotter

Theses

Throughout the 1990s, the incidence of methicillin-resistant Staphylococcus aureus has increased in many health care areas including acute and chronic care facilities, outpatient clinics and in the community. Once introduced into a health care environment, this nosocomial pathogen can spread rapidly and as MRSA are resistant to multiple antibiotics, treatment is often difficult. Therefore effective infection control measures are required to prevent cross-infection and further spread of endemic strains.

In this study, a sensitive and specific triplex-PCR assay was designed for MRSA detection, wherein three genes, the methicillin resistance gene (mecA). femA and the extracellular thermonuclease gene (rmc) were simultaneously …

Development Of An Atomic Force Microscopic Immunosensor For The Ferritin Antigen, Brendan O'Connell

Theses

The adsorption conditions for a range of different immunoreactants to both polystyrene and ultrasmooth silanised silicon surfaces are optimised. The conditions studied were concentration of protein, time and temperature of incubation, pH, molarity and ionic strength of the buffer. The anti-ferritin surface was selected as our model system. This immunosensor was used to develop and evaluate a millispot ELISA for ferritin. The millispot ferritin ELISA developed utilises a sample volume of 6pl and a sample incubation time of 10 minutes. Precision and accuracy studies show the assay to have a dynamic assay range of 7.8-lOOng/ml ferritin. The millispot ELISA detected …

Development And Validation Of Flow-Injection (Continuous-Flow) Elisa Techniques., Declan Gerard Spillane

Theses

A flow-through enzyme-linked immunosorbent assay was developed based on affinity chromatography using the determination of ferritin in serum as a model system. In this method, samples and standards are introduced to separate columns containing immobilised anti-ferritin antibody, and antigen bound by solid-phase antibody is subsequently detected using an anti-ferritin-alkaline phosphatase conjugate. To detect immobilised label, p-nitrophenyl phosphate is added and product is developed in the column at room temperature. Following elution of product from the column, the absorbance is measured and the columns are regenerated using a low pH elution. The final developed system requires approximately 1.5 h for the …