Scattering nephelometry is a particle characterisation method applicable to fluid suspensions containing impurities. Solutions derived by the method feature particle classification by size (diameter), volume or texture as well as continuous on-line and in-situ monitoring, The replacement of turbidimeters with nephelometers in many existing turbidity applications could result in suppression of side effects caused by limitations and uncontrolled parameter drifts and satisfaction of problem-defined constraints at virtually no change in implementation cost. A major issue of nephelometric model design is the selection of a mathematical tool suitable for the modelling of the data analysis system.

This thesis investigates two methods of optical analysis of multiphase fluids. These two methods are nephelometry and the statistical analysis of scattered light intensity fluctuations. Nephelometry is an established technique for investigating particulate suspensions. In this work the basic technique is combined with neural network processing to develop a system capable of automatically distinguishing and quantifYing different suspensions, in particular suspensions of oil. Evidence obtained in this study suggests that neural networks can distinguish the light scattering from suspensions of different size distributions and produce a more accurate estimate of volume fraction than commonly used turbidity measurements. Non-Gaussian, fluctuating light intensities arise from the scattering of light from a varying population of suspended particles. Successful measurement of these intensity fluctuations makes feasible new instrumentation based on the statistical behaviour of the detected signal. Analyses that could prove possible include particle number, size, type and flow characteristics. Photon counting methods have traditionally been used to measure fluctuations from random media but the lower cost of analogue pin diodes makes them the preferred choice of detector if they can be applied usefully. A method of quantifYing the effect of noise from the diode detectors and removing it from the statistics of the fluctuations is developed from a model of the pin diode detectors. Experimental results show that detector noise can be compensated for in the analysis of scattered light fluctuations. Results also indicate that the model used to describe the scattering process is valid and that further work can lead to a practical instrument for the study of suspensions.

Levels of Immunoglobulin G (IgG)-subclasses are analyzed when patients have reoccurring infections and in order to follow the treatment of IgG4 related disease. The measured quantity of IgG-total can be within the reference interval even if the patient has a deficiency in one of the IgG-subclasses. Due to this Sundsvall’s hospital plans to begin analyzing IgG-subclasses. The aim was to verify the performance level of the analysis IgG-subclasses (IgG1-4) with Siemens Atellica NEPH 630. The method was verified by evaluating the method’s precision and by comparing the sum of IgG-subclasses with the quantity of IgG-total analyzed on Cobas c502. In addition, the reference intervals provided by Siemens were evaluated using samples from blood donors. The study evaluated whether there was a correlation between infection with SARS-CoV2 and a deficiency in IgG-subclasses. The verification began by performing quality control twice daily over a period of four weeks. The IgG-subclasses test was performed on blood donor samples with Siemens Atellica NEPH 630 for evaluation of the reference values. The coefficient of variation was less than 6 % for all four subclasses. The reference values for IgG1, IgG2, IgG3, and IgG4 are all in alliance with the reference values provided by Siemens. The sum of IgG-subclasses corresponded well with IgG-total with a correlation value (R) 0.82. No correlation was found between IgG deficiency and infection with SARS-CoV2. The validation of the analysis of IgG-subclasses was successful with quality measurements within the supplier’s intervals. No adjustments will be needed for the reference intervals.

The complement components C3 and C4 are part of the complement system. The complement system has many tasks to perform in the body and consists of several proteins that all have unique tasks. C3 and C4 are examined, for example, if a deficiency is suspected, in the presence of autoimmune diseases or in the case of recurring bacterial infections. When verifying the analysis and after analyzing of external controls, it has been shown that the reference range is falsely high, especially for C3. The study therefore aimed to verify the analysis of C3 and C4 on Atellica NEPH 630 and establish a correct reference range for respective analysis. The method was verified by evaluating the precision of the method and by comparing the concentration of C3 and C4 measured on Cobas 6000 module C500 with the concentration measured on Atellica NEP 630. In addition, the reference intervals provided by Siemens were evaluated using samples from blood donors and from other healthy individuals. The verification began by performing quality controls on three different levels before analysis of samples. The coefficient of variation was less than 5 % for all control levels for both complement analysis. Reference values ​​for C3 were not in alliance with the reference values ​​provided by Siemens. A new proposed range for C3 is 0.48- 1.37 g/L, while the recommended range can be used for C4.

Reumatoid artrit (RA) är den vanligaste autoimmuna sjukdomen och prevalensen är 0.5-1.0% av populationen i industriella länder. Diagnos av RA sker bland annat genom analys av markörerna reumatoid faktor (RF) samt antikroppar mot cykliskt citrullinerad peptid (anti-CCP). Nefelometrisk metod samt CMIA har båda varit huvudmetoder för markörerna RF och anti-CCP respektive vid utredning av RA på Laboratoriemedicin på Länssjukhuset Ryhov i Jönköping. Införskaffning av instrumentet Phadia 250 har gjort det möjligt att sammanställa analyserna för markörerna på samma enhet. Syftet med studien var att verifiera EliA-metoden för anti-CCP samt RF på instrumentet Phadia 250 för att kunna ersätta nuvarande analysmetoder för de båda RA-markörerna. Bestämning av cut-off, mellanliggande precision, inomserieprecision samt överensstämmelse med tidigare metod utfördes. Totalt 115 prover (70 blodgivare, 30 patientprover och 15 konsekutiva prover) användes. En korrelation utfördes för CMIA respektive nefelometriska metoden med EliA-metoden samt en kategoriöverensstämmelse för nefelometrisk metod, CMIA och EliA-metoden. God korrelation erhölls för anti-CCP mellan CMIA och EliA-metoden (r=0.953, p=0.001) samt för RF mellan nefelometriska metoden och EliA-metoden (r=0.835, p=0.048). Analys av samtliga markörer bör inkluderas som screening för RA för att upptäcka sjukdomen. Metoden EliA tillät analys av båda markörer och verifieringen möjliggjorde övergången till EliA-metoden för Laboratoriemedicin i Jönköping. / Rheumatoid arthritis (RA) is the most common autoimmune disease and the prevalence is 0.5-1.0% among the population in industrial countries. Diagnosis of RA is based partially on detection of the autoantibodies rheumatoid factor (RF) and anti-cyclic citrullinated peptide antibodies (anti-CCP). Nephelometry and CMIA have been the main methods for detection of the antibodies at Laboratoriemedicin at the County Hospital Ryhov in Jönköping. The purpose of this study was to verify the EliA-method for anti-CCP and RF on Phadia 250 in order to replace the current methods with the EliA-method. Determination of cut-off, intermediate precision, within-run precision and consistency with the previous method was performed on a total of 115 samples (70 blood-donors, 30 patient samples and 15 consecutive samples). A correlation between CMIA and the nephelometric method with EliA-method was performed and a cathegorical correspondance was done to assess the accordance between the previous methods with the EliA-method. A good correlation was obtained for anti-CCP between CMIA and the EliA-method (r=0.953, p=0.001) and RF obtained good correlation between the nephelometric method and the EliA-method (r=0.835, p=0.048). Analysis of both markers simultaneously has been recommended and the verification enabled the transition to the EliA-method on Phadia 250 for Laboratoriemedicin in Jönköping.

A adesão de plaquetas nas paredes dos vasos sanguíneos e subsequente agregação são eventos cruciais tanto na hemorragia e quanto na trombose. A hiperagregação (agregação excessiva) das plaquetas pode causar a formação de um trombo e a posterior oclusão dos vasos sanguíneos levando a um processo isquêmico. A terapia antiplaquetária com ácido acetilsalicílico reduz em até 25% o risco de infartos do miocárdio não-fatais, acidentes vasculares cerebrais isquêmicos ou mortes de causa vascular em pacientes de alto risco, independentemente do sexo ou idade. Para avaliar laboratorialmente a eficácia dessa terapêutica, o método mais utilizado é o teste de agregação plaquetária, que pode ser feito em duas metodologias, a turbidimétrica e a por impedância elétrica. Com esse cenário, o objetivo do estudo é comparar essas duas metodologias para a monitorização do tratamento. Para isso foram utilizadas amostras de sangue de 30 pacientes adultos (média de 42 anos) de ambos os sexos que fazem uso regular do fármaco e analisadas através das duas metodologias, tendo seus resultados analisados e comparados. Os resultados do estudo mostraram pouca diferença estatística significante (p<0,05) entre os dois métodos nos principais agentes estimulantes, indicando que o método de impedância elétrica pode ser utilizado em rotina laboratorial em substituição, ou em complementação, com a metodologia turbidimétrica para monitorar a terapia antiplaquetária a base de ácido acetilsalicílico / The adhesion of platelets in the blood vessel wall and subsequent aggregation are crucial events in both bleeding and thrombosis. The hyperaggregation (excessive aggregation) of platelets can cause the formation of a thrombus and subsequent occlusion of blood vessels leading to an ischemic process. Antiplatelet therapy with acetylsalicylic acyd reduces by 25% the risk of myocardial infarctions, non-fatal strokes or deaths from vascular causes in patients at high risk, regardless of sex or age. To laboratory evaluation of the effectiveness of this therapy, the method most used is the platelet aggregation test, which can be done in two methods, the turbidimetric and electrical impedance. With this scenario, the objective of the study is to compare these two methodologies for monitoring the treatment. For such purpose, blood samples from 30 adult patients (mean 42 years) of both sexes who make regular use of the drug was analyzed using the two methodologies, and their results was analyzed and compared. The studys results showed little difference statistically significant (p <0.05) between the two methods with the main stimulant agents, indicating that the method of electrical impedance can be used in routine laboratory instead or complementing the methodology of turbidimetric for monitor antiplatelet therapy by acetylsalicylic acyd

A adesão de plaquetas nas paredes dos vasos sanguíneos e subsequente agregação são eventos cruciais tanto na hemorragia e quanto na trombose. A hiperagregação (agregação excessiva) das plaquetas pode causar a formação de um trombo e a posterior oclusão dos vasos sanguíneos levando a um processo isquêmico. A terapia antiplaquetária com ácido acetilsalicílico reduz em até 25% o risco de infartos do miocárdio não-fatais, acidentes vasculares cerebrais isquêmicos ou mortes de causa vascular em pacientes de alto risco, independentemente do sexo ou idade. Para avaliar laboratorialmente a eficácia dessa terapêutica, o método mais utilizado é o teste de agregação plaquetária, que pode ser feito em duas metodologias, a turbidimétrica e a por impedância elétrica. Com esse cenário, o objetivo do estudo é comparar essas duas metodologias para a monitorização do tratamento. Para isso foram utilizadas amostras de sangue de 30 pacientes adultos (média de 42 anos) de ambos os sexos que fazem uso regular do fármaco e analisadas através das duas metodologias, tendo seus resultados analisados e comparados. Os resultados do estudo mostraram pouca diferença estatística significante (p<0,05) entre os dois métodos nos principais agentes estimulantes, indicando que o método de impedância elétrica pode ser utilizado em rotina laboratorial em substituição, ou em complementação, com a metodologia turbidimétrica para monitorar a terapia antiplaquetária a base de ácido acetilsalicílico / The adhesion of platelets in the blood vessel wall and subsequent aggregation are crucial events in both bleeding and thrombosis. The hyperaggregation (excessive aggregation) of platelets can cause the formation of a thrombus and subsequent occlusion of blood vessels leading to an ischemic process. Antiplatelet therapy with acetylsalicylic acyd reduces by 25% the risk of myocardial infarctions, non-fatal strokes or deaths from vascular causes in patients at high risk, regardless of sex or age. To laboratory evaluation of the effectiveness of this therapy, the method most used is the platelet aggregation test, which can be done in two methods, the turbidimetric and electrical impedance. With this scenario, the objective of the study is to compare these two methodologies for monitoring the treatment. For such purpose, blood samples from 30 adult patients (mean 42 years) of both sexes who make regular use of the drug was analyzed using the two methodologies, and their results was analyzed and compared. The studys results showed little difference statistically significant (p <0.05) between the two methods with the main stimulant agents, indicating that the method of electrical impedance can be used in routine laboratory instead or complementing the methodology of turbidimetric for monitor antiplatelet therapy by acetylsalicylic acyd

A cistatina C sérica tem sido apontada como um marcador de filtração glomerular. Neste trabalho realizamos a validação de um método específico e automatizado, a imunonefelometria, mensurando os níveis séricos de cistatina C através do nefelômetro da empresa Behring (BN II). O ensaio perfaz o intervalo de referência de 0.23-7.25 mg/L, até sete vezes acima dos limites considerados normais. A imprecisão intra e interensaio foram de 8,73% and 5,38% , respectively. A recuperação analítica de cistatina C após adição de controle foi entre 86,7 % e 98% (média 92,3%). A estabilidade da cistatina C a temperatura ambiente, sob refrigeração e sob congelamento foi testada. A perda mais significativa foi encontrada nas amostras armazenadas sob temperatura ambiente, onde foi perdido até 10% da concentração inicial. Nós encontramos CV de 14,79 % para sensibilidade analítica. Durante todo o processo nós comparamos os resultados com o controle de qualidade e obtivemos bons resultados. Depois destes testes, nós comparamos as correlações em 3 grupos de pacientes transplantados renais sob diferentes esquemas de imunossupressão (n=197) [azatioprina (n=36), micofenolato mofetil (n=131) ou sirolimus (n=30)], entre as equações de estimativa de filtração glomerular( Cockroft Gault, Nankivell e MDRD) e cistatina C sérica ou creatinina sérica. Nós concluimos que o ensaio nefelométrico cistatina C pode perfeitamente ser adequado à nossa rotina laboratorial e as correlações entre creatinina sérica e as diferentes equações de estimativa de filtração glomerular são melhores do que quando comparamos as mesmas à cistatina C nos 3 grupos independentemente da terapia imunossupressora utilizada. / Serum cystatin C has been suggested as a marker of glomerular filtration rate (GFR). We describe a validation of an automated and rapid particle-enhanced nephelometric immunoassay (PENIA) for measuring serum cystatin C on the Behring nephelometer (BN II). The assay covers the range 0.23-7.25 mg/L, up to seven times the upper limit of normal. The intra- and interassay imprecision are 8,73% and 5,38% , respectively. The analytical recovery by cystatin C addition between 86,7% and 98% (mean 92,3%). The estability of cistatyn C room temperature, refrigerator temperature and frozen temperature was tested. The higher lost was when we stored sample in a room temperature, when we can lost until 10% of initial concentration. We found CV of 14,79 % for analytical sensibility. During all the process we compare the results with a quality control and we obtained good results. After this validation, we have compared the correlation, in 3 different patients groups after renal transplant (n=197) were using different imunossupressors [azatioprine (n=36), micophenolic acid (n=131) or sirolimus (n=30), between glomerular filtration equations (Cockroft Gault, Nankivell and MDRD) and cystatin C or creatinine serum levels. We concluded cystatin C assay may be perfectly used in our laboratory and the correlation between serum creatinine and glomerular filtration equations are better then cystatin C at the same groups independent of imunosupressor therapy.

Les moisissures d’altération sont à l’origine de pertes alimentaires et économiques importantes et certaines espèces peuvent présenter un danger pour la santé humaine et animale avec la production de mycotoxines. Dans ce contexte, la maîtrise de la qualité et de la sécurité des aliments passe par une bonne connaissance des espèces impliquées. Cette connaissance repose sur une identification fiable et rapide et l’obtention d’informations sur les facteurs abiotiques impactant leur développement, tels que les conservateurs, largement utilisés dans l’industrie. Dans ce cadre, les objectifs de thèse étaient de développer l’utilisation de la spectrométrie de masse MALDI-TOF pour l’identification des moisissures et d’évaluer son application à la résolution de complexe d’espèces et au typage, et enfin d’évaluer la néphélométrie laser pour mesurer en haut-débit leur croissance en présence de conservateurs. Dans un premier temps, une base de données robuste a été construite avec près de 6500 spectres correspondant à 136 espèces fongiques. Dans un deuxième temps, la technique MALDI-TOF a été appliquée avec succès à la différenciation de 23 espèces du complexe Aspergillus section Flavi et a permis de différencier des isolats de Penicillium roqueforti appartenant à 3 populations génétiquement différenciées. Enfin, la néphélométrie laser a permis un suivi haut-débit de la croissance de 14 espèces fongiques d’altération en présence de 3 conservateurs et ainsi d’obtenir des informations sur les concentrations minimales inhibitrices de ces derniers. Ces travaux ont démontré l’applicabilité de techniques alternatives permettant d’identifier et de caractériser les moisissures d’altération. / Spoilage fungi represent a major cause of food and economic losses and certain species, which may produce mycotoxins, may also pose a threat to human and animal health. Thus, food safety and quality management relies notably on a good knowledge of the involved species. This knowledge is notably based on their fast and reliable identification and on the study of abiotic factors affecting their growth such as food preservatives, which are commonly used in the food industry. In this context, the objectives of this PhD. thesis were to develop MALDI-TOF mass spectrometry for mold identification and to evaluate its potential for species complex differentiation and strain typing, and finally, to evaluate the use of laser nephelometry to monitor fungal growth in the presence of food preservatives.First, a robust database was developed with 6500 spectra corresponding to 136 spoilage fungi. Then, MALDI-TOF MS was successfully applied to differentiate 23 species of Aspergillus section Flavi and Penicillium roqueforti isolates belonging to 3 genetically distinct populations.Finally, in 14 fungal species, laser nephelometry allowed a high-throughput monitoring of their growth after exposition to 3 different food preservatives and the determination of their associated minimal inhibitory concentrations.Overall, the obtained results demonstrate the usefulness of alternative techniques for identification and characterization of food spoilage fungi.

A cistatina C sérica tem sido apontada como um marcador de filtração glomerular. Neste trabalho realizamos a validação de um método específico e automatizado, a imunonefelometria, mensurando os níveis séricos de cistatina C através do nefelômetro da empresa Behring (BN II). O ensaio perfaz o intervalo de referência de 0.23-7.25 mg/L, até sete vezes acima dos limites considerados normais. A imprecisão intra e interensaio foram de 8,73% and 5,38% , respectively. A recuperação analítica de cistatina C após adição de controle foi entre 86,7 % e 98% (média 92,3%). A estabilidade da cistatina C a temperatura ambiente, sob refrigeração e sob congelamento foi testada. A perda mais significativa foi encontrada nas amostras armazenadas sob temperatura ambiente, onde foi perdido até 10% da concentração inicial. Nós encontramos CV de 14,79 % para sensibilidade analítica. Durante todo o processo nós comparamos os resultados com o controle de qualidade e obtivemos bons resultados. Depois destes testes, nós comparamos as correlações em 3 grupos de pacientes transplantados renais sob diferentes esquemas de imunossupressão (n=197) [azatioprina (n=36), micofenolato mofetil (n=131) ou sirolimus (n=30)], entre as equações de estimativa de filtração glomerular( Cockroft Gault, Nankivell e MDRD) e cistatina C sérica ou creatinina sérica. Nós concluimos que o ensaio nefelométrico cistatina C pode perfeitamente ser adequado à nossa rotina laboratorial e as correlações entre creatinina sérica e as diferentes equações de estimativa de filtração glomerular são melhores do que quando comparamos as mesmas à cistatina C nos 3 grupos independentemente da terapia imunossupressora utilizada. / Serum cystatin C has been suggested as a marker of glomerular filtration rate (GFR). We describe a validation of an automated and rapid particle-enhanced nephelometric immunoassay (PENIA) for measuring serum cystatin C on the Behring nephelometer (BN II). The assay covers the range 0.23-7.25 mg/L, up to seven times the upper limit of normal. The intra- and interassay imprecision are 8,73% and 5,38% , respectively. The analytical recovery by cystatin C addition between 86,7% and 98% (mean 92,3%). The estability of cistatyn C room temperature, refrigerator temperature and frozen temperature was tested. The higher lost was when we stored sample in a room temperature, when we can lost until 10% of initial concentration. We found CV of 14,79 % for analytical sensibility. During all the process we compare the results with a quality control and we obtained good results. After this validation, we have compared the correlation, in 3 different patients groups after renal transplant (n=197) were using different imunossupressors [azatioprine (n=36), micophenolic acid (n=131) or sirolimus (n=30), between glomerular filtration equations (Cockroft Gault, Nankivell and MDRD) and cystatin C or creatinine serum levels. We concluded cystatin C assay may be perfectly used in our laboratory and the correlation between serum creatinine and glomerular filtration equations are better then cystatin C at the same groups independent of imunosupressor therapy.