Background: As a part of a national health control program, Statens VeterinärmedicinskaAnstalt performs diagnostics to screen flocks for certain pathogens causing high mortality,morbidity and/or serious economical losses. There are several viruses in the programincluding IBDV (infectious bursal disease virus), IBV (infectious bronchitis virus) and NDV(Newcastle disease virus). Method: 96 serum samples were collected from different poultryflocks in Sweden and analyzed by ELISA, which are currently used in the health controlprogram as well as by a commercial prototype of a multiplex immunoassay manufactured byLuminex Corp., which is currently under evaluation at the United States Department ofAgriculture USDA. This 4-plex assay detects antibodies for the three above-mentionedviruses as well as antibodies of avian reovirus. In the context of this study the ELISAs run inroutine diagnostics as well as a REO ELISA were used as the standard for comparison.Result: The antibody concentration in serum from vaccinated chickens was high while theantibody concentration level in serum from not vaccinated chickens was low.

The problem of finding targeted medicine is a central problem in chemotherapy. From this point of view the ubiquitin-proteasome system is a highly promising object in the pharmaceutical approach. Proteasome plays a critical role in cellular protein degradation, cell cycle and apoptosis regulation.Proteasome inhibitors are substances blocking the actions of proteasome. Cancer cells are more sensitive to inhibition of the ubiquitin-proteasome system than normal cells. Therefore proteasome inhibitors have the potential to be successfully used in the cancer treatment.The study aimed to test various substances to identify possible proteasome inhibitors with the IncuCyteTM FLR image system and fluorometric microculture cytotoxicity assay.

The ability to detect protein-based biomarkers, which are linked to different diseases like colorectal cancer, is very important as a diagnostic tool. Usually complex biological samples like blood are studied which will contribute to different technical issues when performing an assay. The aim with the project is to optimize and develop the high throughput protein detection technique Proximity Extension Assay, PEA, into a 96-plex panel, in hopes of discovering an expression profile for colorectal cancer. PEA was developed by Olink Bioscience and allows specific proteins in a sample to be quantitatively transformed into nucleic acid sequences that are subsequently detected and quantified with real-time PCR. Two proximity probes containing oligonucleotide sequences bind pairwise to target protein and when brought in proximity, a DNA polymerase will extend a hybridization arm from one probe over to the second forming a double-stranded DNA sequence that can serve as a template in real-time PCR.

Influenza is a viral infection that affects global health and economy with its endemic and sometimes pandemic spread. Rapid detection of Influenza viruses enables antiviral use and can bring financial savings. It is also essential for the global surveillance of prevalent Influenza strains. RT-PCR is considered the most specific and sensitive method for detection of Influenza, but Influenza mutates at a high rate and it is therefore crucial that RT-PCR methods are updated regularly.In 2014, Cepheid released their Xpert Flu/RSV XC assay, which can detect Influenza A and B and RSV by multiplex RT-PCR in approximately one hour. The aim of this study was to evaluate this assay at Laboratoriemedicin Västernorrland by using the laboratory?s previous PCR assay for detection of Influenza viruses as reference method.Real-time RT-PCR was used to compare Xpert Flu/RSV XC to the reference method.

Food-poisoning is a major health problem and an estimated half a million Swedes are food-poisoned annually, with acute gastroenteritis as a consequence. One of the major causes of contaminated foods is related to food- and waterborne viruses. To be able to trace back the source of contaminant, the method of detecting viruses must be specific and sensitive. No standardized method for detecting foods for sapovirus exists today. The aim of the work described in this bachelor thesis is to implement and opti-mize a real-time RT-PCR method for the detection of all genogroups of human sapovirus in foods.

Background: As a part of a national health control program, Statens VeterinärmedicinskaAnstalt performs diagnostics to screen flocks for certain pathogens causing high mortality,morbidity and/or serious economical losses. There are several viruses in the programincluding IBDV (infectious bursal disease virus), IBV (infectious bronchitis virus) and NDV(Newcastle disease virus). Method: 96 serum samples were collected from different poultryflocks in Sweden and analyzed by ELISA, which are currently used in the health controlprogram as well as by a commercial prototype of a multiplex immunoassay manufactured byLuminex Corp., which is currently under evaluation at the United States Department ofAgriculture USDA. This 4-plex assay detects antibodies for the three above-mentionedviruses as well as antibodies of avian reovirus. In the context of this study the ELISAs run inroutine diagnostics as well as a REO ELISA were used as the standard for comparison.Result: The antibody concentration in serum from vaccinated chickens was high while theantibody concentration level in serum from not vaccinated chickens was low.

Background: As a part of a national health control program, Statens VeterinärmedicinskaAnstalt performs diagnostics to screen flocks for certain pathogens causing high mortality,morbidity and/or serious economical losses. There are several viruses in the programincluding IBDV (infectious bursal disease virus), IBV (infectious bronchitis virus) and NDV(Newcastle disease virus). Method: 96 serum samples were collected from different poultryflocks in Sweden and analyzed by ELISA, which are currently used in the health controlprogram as well as by a commercial prototype of a multiplex immunoassay manufactured byLuminex Corp., which is currently under evaluation at the United States Department ofAgriculture USDA. This 4-plex assay detects antibodies for the three above-mentionedviruses as well as antibodies of avian reovirus. In the context of this study the ELISAs run inroutine diagnostics as well as a REO ELISA were used as the standard for comparison.Result: The antibody concentration in serum from vaccinated chickens was high while theantibody concentration level in serum from not vaccinated chickens was low.

Abstract Tripeptidyl-peptidase II (TPPII), is present in most eukaryotic cells. It cuts tripeptides from the N-terminus of peptides and is especially important for degrading peptides longer than 15 amino acids. TPPII also tailors long peptides into suitable substrates for the enzymes which transport and produce the peptides that MHC I present. Increased levels of TPPII have also been found in certain cancer cells, thus it is of interest to determine if TPPII could be used as a tumour marker.The aim of this study was to optimise and evaluate 3 different methods for quantifying TPPII. Western blot, enzyme-linked immunosorbent assay (ELISA) and fluorophore-linked immunosorbent assay (FLISA) protocols were optimised regarding incubation times and antibody dilutions.

Background: Hepatitis E (HEV) is a small, non-enveloped virus that belongs to the viral genus Hepevirus. HEV is a positive sense single-stranded RNA virus and there is insufficient information regarding its replication. This is mainly because the virus has low capacity to grow in normally used cell cultures. Many specific strand assay detection studies have been done in order to understand more about HEV replication. Unfortunately, these assays have the disadvantage of giving false positive results.Aim: The aim of this project was to improve the positive strand assay to increase specificity and eliminate false positivity which is due to high sensitivity of the polymerase chain reaction (PCR).

The analysis of acute phase protein serum amyloid A (SAA) has recently been brought into clinical use in veterinary medicine. Some of the difficulties with incorporating the SAA method in clinical practice have been the expensive and rather large equipment required for the method. Due to these difficulties only larger clinics can afford to use the SAA analysis.The company Equinostic has recently developed a smaller instrument that costs one-tenth of a larger instrument. The instrument is named EVA1 and has so far only been used to analyze SAA in horses.The aim of this study was to investigate if the EVA1 instrument could be used to analyze SAA in cats. This study included 24 serum samples from cat, which were first analyzed twice on the EVA1 instrument and then sent to the Strömsholm Referral Animal Hospital in Sweden where they reanalyzed the samples using a validated reference method.

The Annexin-V/propidium iodide (AV/PI) assay detects early membrane changes in spermatozoa and viability using flow cytometry. It is an objective method that evaluates a much larger number of single spermatozoa compared with the usual estimation of motility, the most commonly used method for routine sperm evaluation. In the present study four ejaculates from four stallions were used in a split-sample design to investigate whether and how the number of intact spermatozoa is affected over time and after incubation at different temperatures, either those recorded during shipment of stallion semen for AI (9 or 5°C) or room temperature (20°C). The Annexin-V results did not reflect the motility results that were monitored in parallel. While motility decreased over time, the number of spermatozoa with intact membranes (viable) remained unchanged for at least 21 hours following a short period of initial instability.

A plant's defensive ability against herbivore attack can be influenced by many different factors, one of them being domestication. During human selection to improve the yield of plants, the resistance against herbivore attack can have been lowered, due to a trade-off for use of resources between these traits in the plants. In this thesis I investigated the effect of domestication on resistance against herbivorous insects in cotton plants. I used different varieties and species of wild and domesticated cotton in three different experiments. A feeding assay was conducted, the mortality, development time and pupal weight of larvae of the Egyptian cotton leaf worm Spodoptera littoralis was studied. In a second experiment the preference of the root-feeding beetle Agriotes spp.

Serum thymidine kinase 1 (TK1) activity is used as a tumor marker in disease monitoring in veterinary and human medicine. TK1, an intracellular enzyme, is involved in a salvage pathway of DNA precursor synthesis. TK1 is used in DNA precursor production by catalyzing the transfer of the gamma-phosphate-group from a phosphate-donor to the 5?- hydroxyl-group of thymidine forming thymidine-monophosphate. Nucleoside monophosphosphates are finally converted into thymidine-triphosphates. TK1 activity significantly rises in the G1 and the S phase of the cell cycle.

Cardiac troponin I (cTnI) is considered a specific and a sensitive biomarker of cardiac disease, and due to a high inter-species sequence homology; human cTnI assays can often be used in veterinary medicine. The main aim of this study was to perform an initial evaluation of a human cTnI test, the Meritas Troponin I test, for measurement of equine plasma cTnI, in order to investigate whether the test could be a possible subject for a more comprehensive validation. The hypothesis was that the test could measure equine cTnI, as there is a good homology between equine cTnI and human cTnI. The evaluation included a dilution parallelism, an intra-assay precision study and measurement of plasma cTnI healthy horses. In the dilution parallelism and the intra-assay precision study, equine plasma with previously established high cTnI concentration, according to another cTnI assay, was used. In addition, cTnI concentration was measured in plasma, collected from 19 horses, without signs of disease at physical and ECG examinations. The obtained curve in the dilution parallelism was linear, indicating that the Meritas Troponin I test can be used to measure different concentrations of equine cTnI in plasma.

The genus Campylobacter is globally recognised as the leading bacterial cause of human foodborne gastroenteritis. Every year around 8000 Swedes are infected by Campylobacter. Most people are infected by thermotolerant Campylobacter species, commonly C. jejuni and C. coli.