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.

Vitamin D är den sammanlagda koncentrationen för metaboliterna vitamin D2 och vitamin D3 och är en tillförlitlig indikator gällande vitamin D-statusen i kroppen. Den huvudsakliga källan till vitamin D är bildandet av metaboliten vitamin D3 i huden med hjälp av ultraviolett B-ljus och vitaminet har bland annat en viktig roll för skelettets utveckling. Koncentrationen kalcidiol (25(OH) vitamin D) kan bestämmas till exempel med hjälp av elektrokemiluminiscens med kompetitiv inbindning. Mätprincipen har flera likheter med kemiluminiscens med den huvudsakliga skillnaden att elektrisk spänning används för att starta reaktionen.   När en analys ska flyttas från ett instrument till ett annat är det viktigt att verifiera att de är överensstämmande gällande bland annat precision. Syftet med projektet var att genomföra en metodjämförelse för analys av kalcidiol i serum (S-Kalcidiol) mellan instrumenten IDS-iSYS och Cobas E801 samt att genomföra en jämförelse av kontroller från företagen Roche Diagnostics och ThermoFisher Scientific och utifrån de erhållna resultaten avgöra vilka som var mest lämpliga att använda vid analys med Cobas E801. Kontrollerna från Roche Diagnostics är optimerade för analysen, men det vore fördelaktigt att kunna använda kontrollerna från ThermoFisher Scientific eftersom de även kan användas för flera andra analyser. Metodjämförelsen gjordes med hjälp av en inomserie- och totalimprecisionsstudie med en låg och en hög kontrollnivå från företagen Roche Diagnostics och ThermoFisher Scientific samt en korrelationsstudie med 20 stycken patientprover bestående av serum.  Inomserie- och totalimprecisionen resulterade i att Cobas E801 hade likvärdig precision med IDS-iSYS. Gällande kontrollerna var endast kontrollerna från Roche Diagnostics inom acceptansintervallen och är därmed lämpligast att använda. Korrelationsstudien resulterade i determinationskoefficienten 0,942, vilket tyder på ett tydligt linjärt samband. Studien visar dock att Cobas E801 har en större spridning när koncentrationen kalcidiol överstiger 50 nmol/L.  Utifrån de erhållna resultaten kan en flytt av analysen från IDS-iSYS till Cobas E801 genomföras för att effektivisera arbetsflödet.

Introdução A preocupação crescente com as possíveis consequências da replicação viral no sistema nervoso central mostra a necessidade da detecção do HIV no compartimento cerebral. O teste de PCR em tempo real desenvolvido pela Abbott, o Abbott m2000 RealTime HIV-1 (m2000rt), quantifica a carga viral do HIV em amostras de sangue com um procedimento efetivo e de baixo custo no nosso país, por isso é adotado como método padrão pelo Ministério da Saúde, mas não é utilizado em amostras de líquor. O ensaio produzido pela Roche, o COBAS TaqMan HIV-1, version 2 (COBAS v2.0), é o método de PCR em tempo real que tem sido amplamente utilizado para detectar a carga viral do HIV no compartimento cerebral. No entanto, esse método ainda não foi validado para esse propósito e seu custo pode ser uma limitação em diversas regiões com baixos recursos. Objetivos Considerando que não há uma metodologia padronizada para essa situação específica (detecção do HIV no líquor), nós conduzimos esse estudo a fim comparar os desempenhos dos testes m2000rt e COBAS v2.0, na tentativa de propôr um método alternativo e com baixos custos ao mais utilizado nesse contexto (COBAS v2.0). Métodos O estudo foi realizado no período de maio de 2015 a julho de 2016. O cálculo do tamanho da amostra foi baseado em dados de um estudo piloto que revelaram ser necessário um número mínimo de 37 amostras, para detectar uma diferença de 0,20 log10 na carga viral, com um coeficiente de correlação de 0,979 e um poder de 90%. Essa equação permitiria uma perda de 10%. As amostras de líquor foram coletadas consecutivamente a partir de 37 pacientes HIV positivos atendidos no Hospital de Clínicas de Porto Alegre/RS. Os métodos foram processados de acordo com o proposto pelo fabricante para utilização com amostras de plasma. Pequenas modificações foram necessárias no teste em estudo (m2000rt) para neutralizar qualquer diferença metodológica e evitar vieses de mensuração: o congelamento das amostras foi realizado a -20ºC até o momento da análise. O ensaio COBAS v2.0 foi utilizado como referência, uma vez que é o método mais utilizado. Foram realizadas análises quantitativas com resultados que estavam dentro da faixa linear em ambos os métodos (n = 18). Para tornar os métodos comparáveis, adotou-se o limite de detecção do ensaio m2000rt para ambos (40 cp/mL ou 1,60 log10 cp/mL). Os resultados abaixo do limite de detecção foram apresentados como uma variável categórica, uma vez que não são quantificáveis. O coeficiente de correlação de Pearson foi utilizado para comparar os métodos. A normalidade das variáveis foi então resumida calculando o viés estimado pela diferença média "đ " e o desvio padrão das diferenças realizadas pelo teste t para amostras pareadas. Com base na falta de normalidade dos métodos, o grau de concordância dos resultados das cargas virais de HIV foi analisado pelo índice Kappa. Esse estudo foi aprovado pelo comitê de ética do Hospital de Clínicas de Porto Alegre (RS)/ Brasil, registrado na Plataforma Brasil como sendo CAAE: 35072214.7.0000.532. Conclusão Em conclusão, o teste m2000rt que foi modificado para este ensaio mostrou boa concordância e correlação com o teste mais utilizado nesse contexto e pode ser considerado um método alternativo com resultados semelhantes ao COBAS v2.0 e baixos custos na quantificação da carga viral do HIV no líquor. Sugerimos, principalmente em locais onde este método está prontamente disponível com uma relação custo-benefício aceitável, que o exame m2000rt deva ser realizado. / Introduction Growing concern about possible consequences of viral replication in the central nervous system shows the need for HIV detection in the cerebral compartment. The real time PCR test developed by Abbott, Abbott RealTime m2000 HIV-1 (m2000rt) quantifies HIV viral load in blood samples effectively and with low costs Brazil. It is the standard method by the Brazilian Ministry of Health, but it has never been utilized to measure HIV in cerebrospinal fluid samples. The assay produced by Roche, COBAS TaqMan HIV-1, version 2 (COBAS v2.0) is the real-time PCR method that has been widely used to detect HIV viral load in cerebral compartment. However, this method has not yet been validated for this purpose and its cost may be a limitation in several regions in the world with low resources. Objective Taking under consideration that there was no standard methodology for this specific situation (detecting HIV in cerebrospinal fluid), we conducted this study to compare the performances of the m2000rt and COBAS v2.0 assays, to propose an alternative and low-cost method to more used in this context (COBAS v2.0). Methods The study was conducted from May 2015 to July 2016. The sample size calculation was based on data from a pilot trial that revealed that a minimum of 37 samples would be needed to detect a difference of 0.20 log10 in viral load, with a correlation coefficient of 0.979 and a 90% power. This equation would allow a 10% lost. CSF samples were collected consecutively from 37 HIV-positive patients seen at Hospital de Clínicas, Porto Alegre, RS. Methods were processed according to proposed by the manufacturer for utilization with plasma samples. Small modifications were necessary in the study test (m2000rt) to neutralize any methodological differences, thus avoiding measurement bias: the freezing of samples was carried out at -20ºC until the moment of the analysis. The COBAS v2.0 test was used as a reference since it is the most commonly used method. Quantitative analyzes were performed with results that were within the linear range in both methods (n=18). To make the methods comparable, the detection limit of the m2000rt assay for both (40 cp/mL or 1.60 log10 cp/mL) was adopted. The results below the limit of detection were presented as a categorical variable, since they are not quantifiable. The Pearson correlation coefficient was used to compare methods. The normality of the variables was then summarized calculating the estimated bias by the mean difference "đ" and standard deviation of the differences performed by t test for paired samples. Based on the lack of normality of the methods, the degree of agreement of the HIV viral load results was analyzed by the Kappa index. This study was approved by the Hospital de Clínicas of Porto Alegre (southern Brazil) Ethics Review Board, registered in the Brazil Platform as CAAE: 35072214.7.0000.532. Conclusion In conclusion, the m2000rt test that was modified for this trial showed good agreement and correlation with the most used test in this context and can be considered an alternative method with similar results to COBAS v2.0 and low costs in the HIV viral load quantification in cerebrospinal fluid. We suggest, especially in places where this method is readily available with an acceptable cost-benefit ratio, that the m2000rt exam should be performed.

Introdução A preocupação crescente com as possíveis consequências da replicação viral no sistema nervoso central mostra a necessidade da detecção do HIV no compartimento cerebral. O teste de PCR em tempo real desenvolvido pela Abbott, o Abbott m2000 RealTime HIV-1 (m2000rt), quantifica a carga viral do HIV em amostras de sangue com um procedimento efetivo e de baixo custo no nosso país, por isso é adotado como método padrão pelo Ministério da Saúde, mas não é utilizado em amostras de líquor. O ensaio produzido pela Roche, o COBAS TaqMan HIV-1, version 2 (COBAS v2.0), é o método de PCR em tempo real que tem sido amplamente utilizado para detectar a carga viral do HIV no compartimento cerebral. No entanto, esse método ainda não foi validado para esse propósito e seu custo pode ser uma limitação em diversas regiões com baixos recursos. Objetivos Considerando que não há uma metodologia padronizada para essa situação específica (detecção do HIV no líquor), nós conduzimos esse estudo a fim comparar os desempenhos dos testes m2000rt e COBAS v2.0, na tentativa de propôr um método alternativo e com baixos custos ao mais utilizado nesse contexto (COBAS v2.0). Métodos O estudo foi realizado no período de maio de 2015 a julho de 2016. O cálculo do tamanho da amostra foi baseado em dados de um estudo piloto que revelaram ser necessário um número mínimo de 37 amostras, para detectar uma diferença de 0,20 log10 na carga viral, com um coeficiente de correlação de 0,979 e um poder de 90%. Essa equação permitiria uma perda de 10%. As amostras de líquor foram coletadas consecutivamente a partir de 37 pacientes HIV positivos atendidos no Hospital de Clínicas de Porto Alegre/RS. Os métodos foram processados de acordo com o proposto pelo fabricante para utilização com amostras de plasma. Pequenas modificações foram necessárias no teste em estudo (m2000rt) para neutralizar qualquer diferença metodológica e evitar vieses de mensuração: o congelamento das amostras foi realizado a -20ºC até o momento da análise. O ensaio COBAS v2.0 foi utilizado como referência, uma vez que é o método mais utilizado. Foram realizadas análises quantitativas com resultados que estavam dentro da faixa linear em ambos os métodos (n = 18). Para tornar os métodos comparáveis, adotou-se o limite de detecção do ensaio m2000rt para ambos (40 cp/mL ou 1,60 log10 cp/mL). Os resultados abaixo do limite de detecção foram apresentados como uma variável categórica, uma vez que não são quantificáveis. O coeficiente de correlação de Pearson foi utilizado para comparar os métodos. A normalidade das variáveis foi então resumida calculando o viés estimado pela diferença média "đ " e o desvio padrão das diferenças realizadas pelo teste t para amostras pareadas. Com base na falta de normalidade dos métodos, o grau de concordância dos resultados das cargas virais de HIV foi analisado pelo índice Kappa. Esse estudo foi aprovado pelo comitê de ética do Hospital de Clínicas de Porto Alegre (RS)/ Brasil, registrado na Plataforma Brasil como sendo CAAE: 35072214.7.0000.532. Conclusão Em conclusão, o teste m2000rt que foi modificado para este ensaio mostrou boa concordância e correlação com o teste mais utilizado nesse contexto e pode ser considerado um método alternativo com resultados semelhantes ao COBAS v2.0 e baixos custos na quantificação da carga viral do HIV no líquor. Sugerimos, principalmente em locais onde este método está prontamente disponível com uma relação custo-benefício aceitável, que o exame m2000rt deva ser realizado. / Introduction Growing concern about possible consequences of viral replication in the central nervous system shows the need for HIV detection in the cerebral compartment. The real time PCR test developed by Abbott, Abbott RealTime m2000 HIV-1 (m2000rt) quantifies HIV viral load in blood samples effectively and with low costs Brazil. It is the standard method by the Brazilian Ministry of Health, but it has never been utilized to measure HIV in cerebrospinal fluid samples. The assay produced by Roche, COBAS TaqMan HIV-1, version 2 (COBAS v2.0) is the real-time PCR method that has been widely used to detect HIV viral load in cerebral compartment. However, this method has not yet been validated for this purpose and its cost may be a limitation in several regions in the world with low resources. Objective Taking under consideration that there was no standard methodology for this specific situation (detecting HIV in cerebrospinal fluid), we conducted this study to compare the performances of the m2000rt and COBAS v2.0 assays, to propose an alternative and low-cost method to more used in this context (COBAS v2.0). Methods The study was conducted from May 2015 to July 2016. The sample size calculation was based on data from a pilot trial that revealed that a minimum of 37 samples would be needed to detect a difference of 0.20 log10 in viral load, with a correlation coefficient of 0.979 and a 90% power. This equation would allow a 10% lost. CSF samples were collected consecutively from 37 HIV-positive patients seen at Hospital de Clínicas, Porto Alegre, RS. Methods were processed according to proposed by the manufacturer for utilization with plasma samples. Small modifications were necessary in the study test (m2000rt) to neutralize any methodological differences, thus avoiding measurement bias: the freezing of samples was carried out at -20ºC until the moment of the analysis. The COBAS v2.0 test was used as a reference since it is the most commonly used method. Quantitative analyzes were performed with results that were within the linear range in both methods (n=18). To make the methods comparable, the detection limit of the m2000rt assay for both (40 cp/mL or 1.60 log10 cp/mL) was adopted. The results below the limit of detection were presented as a categorical variable, since they are not quantifiable. The Pearson correlation coefficient was used to compare methods. The normality of the variables was then summarized calculating the estimated bias by the mean difference "đ" and standard deviation of the differences performed by t test for paired samples. Based on the lack of normality of the methods, the degree of agreement of the HIV viral load results was analyzed by the Kappa index. This study was approved by the Hospital de Clínicas of Porto Alegre (southern Brazil) Ethics Review Board, registered in the Brazil Platform as CAAE: 35072214.7.0000.532. Conclusion In conclusion, the m2000rt test that was modified for this trial showed good agreement and correlation with the most used test in this context and can be considered an alternative method with similar results to COBAS v2.0 and low costs in the HIV viral load quantification in cerebrospinal fluid. We suggest, especially in places where this method is readily available with an acceptable cost-benefit ratio, that the m2000rt exam should be performed.

Introdução A preocupação crescente com as possíveis consequências da replicação viral no sistema nervoso central mostra a necessidade da detecção do HIV no compartimento cerebral. O teste de PCR em tempo real desenvolvido pela Abbott, o Abbott m2000 RealTime HIV-1 (m2000rt), quantifica a carga viral do HIV em amostras de sangue com um procedimento efetivo e de baixo custo no nosso país, por isso é adotado como método padrão pelo Ministério da Saúde, mas não é utilizado em amostras de líquor. O ensaio produzido pela Roche, o COBAS TaqMan HIV-1, version 2 (COBAS v2.0), é o método de PCR em tempo real que tem sido amplamente utilizado para detectar a carga viral do HIV no compartimento cerebral. No entanto, esse método ainda não foi validado para esse propósito e seu custo pode ser uma limitação em diversas regiões com baixos recursos. Objetivos Considerando que não há uma metodologia padronizada para essa situação específica (detecção do HIV no líquor), nós conduzimos esse estudo a fim comparar os desempenhos dos testes m2000rt e COBAS v2.0, na tentativa de propôr um método alternativo e com baixos custos ao mais utilizado nesse contexto (COBAS v2.0). Métodos O estudo foi realizado no período de maio de 2015 a julho de 2016. O cálculo do tamanho da amostra foi baseado em dados de um estudo piloto que revelaram ser necessário um número mínimo de 37 amostras, para detectar uma diferença de 0,20 log10 na carga viral, com um coeficiente de correlação de 0,979 e um poder de 90%. Essa equação permitiria uma perda de 10%. As amostras de líquor foram coletadas consecutivamente a partir de 37 pacientes HIV positivos atendidos no Hospital de Clínicas de Porto Alegre/RS. Os métodos foram processados de acordo com o proposto pelo fabricante para utilização com amostras de plasma. Pequenas modificações foram necessárias no teste em estudo (m2000rt) para neutralizar qualquer diferença metodológica e evitar vieses de mensuração: o congelamento das amostras foi realizado a -20ºC até o momento da análise. O ensaio COBAS v2.0 foi utilizado como referência, uma vez que é o método mais utilizado. Foram realizadas análises quantitativas com resultados que estavam dentro da faixa linear em ambos os métodos (n = 18). Para tornar os métodos comparáveis, adotou-se o limite de detecção do ensaio m2000rt para ambos (40 cp/mL ou 1,60 log10 cp/mL). Os resultados abaixo do limite de detecção foram apresentados como uma variável categórica, uma vez que não são quantificáveis. O coeficiente de correlação de Pearson foi utilizado para comparar os métodos. A normalidade das variáveis foi então resumida calculando o viés estimado pela diferença média "đ " e o desvio padrão das diferenças realizadas pelo teste t para amostras pareadas. Com base na falta de normalidade dos métodos, o grau de concordância dos resultados das cargas virais de HIV foi analisado pelo índice Kappa. Esse estudo foi aprovado pelo comitê de ética do Hospital de Clínicas de Porto Alegre (RS)/ Brasil, registrado na Plataforma Brasil como sendo CAAE: 35072214.7.0000.532. Conclusão Em conclusão, o teste m2000rt que foi modificado para este ensaio mostrou boa concordância e correlação com o teste mais utilizado nesse contexto e pode ser considerado um método alternativo com resultados semelhantes ao COBAS v2.0 e baixos custos na quantificação da carga viral do HIV no líquor. Sugerimos, principalmente em locais onde este método está prontamente disponível com uma relação custo-benefício aceitável, que o exame m2000rt deva ser realizado. / Introduction Growing concern about possible consequences of viral replication in the central nervous system shows the need for HIV detection in the cerebral compartment. The real time PCR test developed by Abbott, Abbott RealTime m2000 HIV-1 (m2000rt) quantifies HIV viral load in blood samples effectively and with low costs Brazil. It is the standard method by the Brazilian Ministry of Health, but it has never been utilized to measure HIV in cerebrospinal fluid samples. The assay produced by Roche, COBAS TaqMan HIV-1, version 2 (COBAS v2.0) is the real-time PCR method that has been widely used to detect HIV viral load in cerebral compartment. However, this method has not yet been validated for this purpose and its cost may be a limitation in several regions in the world with low resources. Objective Taking under consideration that there was no standard methodology for this specific situation (detecting HIV in cerebrospinal fluid), we conducted this study to compare the performances of the m2000rt and COBAS v2.0 assays, to propose an alternative and low-cost method to more used in this context (COBAS v2.0). Methods The study was conducted from May 2015 to July 2016. The sample size calculation was based on data from a pilot trial that revealed that a minimum of 37 samples would be needed to detect a difference of 0.20 log10 in viral load, with a correlation coefficient of 0.979 and a 90% power. This equation would allow a 10% lost. CSF samples were collected consecutively from 37 HIV-positive patients seen at Hospital de Clínicas, Porto Alegre, RS. Methods were processed according to proposed by the manufacturer for utilization with plasma samples. Small modifications were necessary in the study test (m2000rt) to neutralize any methodological differences, thus avoiding measurement bias: the freezing of samples was carried out at -20ºC until the moment of the analysis. The COBAS v2.0 test was used as a reference since it is the most commonly used method. Quantitative analyzes were performed with results that were within the linear range in both methods (n=18). To make the methods comparable, the detection limit of the m2000rt assay for both (40 cp/mL or 1.60 log10 cp/mL) was adopted. The results below the limit of detection were presented as a categorical variable, since they are not quantifiable. The Pearson correlation coefficient was used to compare methods. The normality of the variables was then summarized calculating the estimated bias by the mean difference "đ" and standard deviation of the differences performed by t test for paired samples. Based on the lack of normality of the methods, the degree of agreement of the HIV viral load results was analyzed by the Kappa index. This study was approved by the Hospital de Clínicas of Porto Alegre (southern Brazil) Ethics Review Board, registered in the Brazil Platform as CAAE: 35072214.7.0000.532. Conclusion In conclusion, the m2000rt test that was modified for this trial showed good agreement and correlation with the most used test in this context and can be considered an alternative method with similar results to COBAS v2.0 and low costs in the HIV viral load quantification in cerebrospinal fluid. We suggest, especially in places where this method is readily available with an acceptable cost-benefit ratio, that the m2000rt exam should be performed.

Cancerassocierade antigen (CA) används vid diagnostisering, prognosbestämning och för att följa behandling vid cancer. Exempel på CA är CA 15–3, CA 19–9 och CA 125. Syftet med arbetet var att undersöka hållbarheten för blodprover förvarade i primärrör som skulle analyseras för CA 15–3, CA 19–9 och CA 125 och jämföra med nuvarande metod på klinisk kemi, Oskarshamn, Region Kalmar län. Enligt nuvarande metod överförs plasman till nya provrör innan analys. Metoden som användes var att dubbelprov togs och det ena förvarades i primärrör och analyserades på Roche Cobas 411® tre, fem och tio dagar efter provtagning. Det andra provet överfördes till nytt provrör inför analys. Den procentuella skillnaden mellan analys av CA i primärrör och överfört prov beräknades. För alla tre analyserna var den procentuella skillnaden störst hos de lägsta koncentrationerna av CA. Medelvärde för analyserna av CA, standardavvikelse (SD) och variationskoefficient beräknades. SD för CA jämfördes mot kontrollerna MAS Omni Immune Pro 1 och MAS Omni Immune Pro 3s SD. Provernas SD var mindre för kontrollerna förutom för CA 19–9 >1100 kIE/L. I den gruppen analyserades dock för få analyser för att utföra statistisk analys och gav därmed ingen slutsats hur CA 19–9 påverkades av de olika förvaringarna. För övriga analyser och för CA 19–9 i lägre koncentrationer kunde slutsatsen dras att prover kunde förvaras i primärrör i tio dagar utan att analysresultatet påverkades. / Cancer-associated antigens (CA) are used for diagnosis, establish prognosis and to monitor treatment of cancer. Three CA are CA 15-3, CA 19-9 and CA 125. The aim of this study was to evaluate the stability of CA 15-3, CA 19-9 and CA 125 blood samples in primary tubes coated with Litium-heparin gel and compare the results with the method used at Klinisk Kemi Oskarshamn, Region Kalmar where the samples today are transferred to new tubes before analysis. In this study two samples were collected from each patient, one was stored and analyzed according to the current method. The other sample was stored in the primary tube and analyzed three, five and ten days after collection. Roche Cobas 411 instrument was used for analysis. The mean value for the samples in primary tubes was compared to the current method and were calculated in percent. The difference was more distinct when the concentration of the analyte was low. The mean value, standard deviation (SD) and variation coefficient were calculated for each analyte. The SD of the samples were compared to the SD of the controls IM 1 and IM3. All the obtained SD from the samples were lower than the control SD, except for CA 19-9 > 1100 kIE/L. Unfortunately, there were only three test results in the concentration interval and therefore no statistic conclusion could be made. The conclusion for the other analytes CA 15-3, CA 125 and CA 19-9 in lower concentrations was that the storage in primary tubed up to ten days did not affect the test results.

Biotin är ett vitamin som finns naturligt i livsmedel och det dagliga intaget nås via födan. Höga doser biotintillskott samt höga doser biotin i läkemedel, kan leda till biotininterferens i kliniska immunokemiska analyser. Roche Diagnostics® vill införa nya reagens med högre tröskel för biotininterferens för att minska risken för biotininterferens vid analys av patientplasma. Därför var syftet med studien att metodverifiera fyra nya reagens från Roche Diagnostics® som används vid diagnostisering och behandling av hjärtsvikt, sepsis, och prostatacancer. De fyra reagensen, Elecsys® proBNP II, Elecsys® BRAHMS PCT, Elecsys® total PSA samt Elecsys® free PSA metodverifierades för att användas på Cobas® e801. Studiematerialet bestod av 20 patientprover av litiumheparinplasma per reagens (totalt 80 patientprover). Resultatet av verifieringen av Elecsys® proBNP II visade en korrelation till det befintliga reagenset på r = 0,9998 och Bland-Altman analys visade en spridning av resultaten på < 10 %; inomserieprecisionsstudien gav CV 1,56 %. Elecsys® BRAHMS PCT hade en korrelation på r = 0,9997 och Bland-Altman analysen visade en spridning på > 10 %; inomserieprecisionsstudien gav CV 1,70 %. För Elecsys® total PSA och free PSA fanns korrelationen till det befintliga reagenset på r = 1 respektive 0,9997 och Bland- Altman analysen visade en spridning på < 10 % hos båda reagensen. Inomserieprecisionsstudien gav CV 0,44 % respektive CV 2,67 %. Resultaten för samtliga reagens uppvisar god korrelation till det befintliga reagenset och en hög mätnoggrannhet vilket talar för att de fyra nya reagensen kan tas i bruk. / Biotin is naturally found in foods, and we obtain this vitamin through our daily diet. Biotin supplements as well as high doses of biotin in drugs can lead to biotin interference in clinical immunochemical analyzes. Therefore, the purpose of this study was to methodically verify four new reagents from Roche Diagnostics® with a higher threshold for biotin interference, used in the diagnosis and treatment of heart faliure, sepsis and prostate cancer. The four reagents, Elecsys® proBNP II, Elecsys® BRAHMS PCT, Elecsys® total PSA and Elecsys® free PSA were method-verified for use on Cobas® e801. The study material consisted of 20 patient samples of lithium heparin plasma per reagent. In total 80 samples were analyzed.The result of the verification of Elecsys® proBNP II showed a correlation to the existing reagent of r = 0.9998 and Bland-Altman analysis showed a distribution of the results of <10 %. The withinseries precision study yielded CV 1.56 %. Elecsys® BRAHMS PCT had a correlation of r = 0.9997 and the Bland-Altman analysis showed a distribution of > 10 %. The withinseries precision study gave CV 1.70 %. For Elecsys® total PSA and free PSA, the correlation to the existing reagent was r = 1 and 0.9997, respectively, and the Bland-Altman analysis showed a distribution of <10 % in both reagents. The withinseries precision study yielded CV 0.44 % and CV 2.67 % respectively.The results for all reagents show a good correlation to the existing reagent and a high accuracy of measurement, which indicates that the four new reagents can be used.

Etylenglykol (etan-1,2-diol) är en dihydroxyalkohol som är en komponent i kylarvätska och andra frost- och kylskyddsmedel. Förtäring av etylenglykol leder till allvarliga skador och i värsta fall död utan behandling. I Sverige år 2020 fanns det endast ett tiotal laboratorier som erbjöd analys av etylenglykol dygnet runt. Detta leder till att prover ofta behöver skickas till större laboratorier med taxi vilket i sin tur leder till en försenad diagnosticering av patienten. Syftet med examensarbetet var att verifiera en enzymatisk kolorimetrisk metod för analys av etylenglykol i plasma på instrumentet Roche Cobas® 8000 modul c502. Reagenset som verifierades var DiscretPak™ Ethylene Glycol Reagents från företaget Catachem. Verifieringen gjordes med avseende på total precision (repeterbarhet), inomserieprecision och linjäritet. Resultaten jämfördes med analys på gaskromatograf. Provmaterialet bestod av patientprover av litiumheparinplasma,  patientprover av serum och kontrollprover från Equalis. Resultatet som erhölls vid verifieringen visade på linjär korrelation mellan den enzymatiska metoden och GC-analys. En negativ bias observerades dock i jämförelse med analys på gaskromatograf. Utvärdering av repeterbarhet gav CV 4,6% vid 9,0 mmol/L och 3,66% vid 40,0 mmol/L. Inomserieprecisionstudie gav CV 14,7% vid 1 mmol/L, 5,2% vid 4 mmol/L och 1,4% vid 17 mmol/L. Precisionen är viktigast vid de lägre koncentrationerna. Insättning av behandling med antidot är aktuellt vid 4 mmol/L. Utvärdering av linjäritet visade på ett starkt linjärt samband hos analysen vid koncentrationer <50 mmol/L. Vid koncentrationer >50 mmol/L fanns ett linjärt samband men en minskad överensstämmelse mellan den beräknade och den uppmätta koncentrationen observerades. Metodverifieringen ansågs vara godkänd för kliniskt bruk och analysen kommer att införas i analyssortimentet hos Klinisk Kemi i Växjö. / Ethylene glycol is an alcohol that is a common component in antifreeze. Ingestion of ethylene glycol will, without treatment, lead to severe organ damage and in worst-case death. In 2020 there was only a few laboratories in Sweden that offered analysis of ethylene glycol all hours of the day and week. This means that samples often need to be transported to laboratories at larger hospitals which leads delayed diagnosis of the patient. The purpose of this study was to verify an enzymatic method for analysis of ethylene glycol in plasma on the instrument Roche Cobas® 8000 module c502. The reagent that was used was DiscretPak™ Ethylene Glycol Reagents from Catachem. The study included evaluation of total precision, within-run precision, linearity, and a comparison with analysis with gas chromatography (GC). The sample material consisted of patient samples of plasma, patient samples of serum and quality controls from Equalis. The result of the study showed linear correlation between the enzymatic method and analysis with GC. A negative bias was observed in comparison to analysis with GC. The coefficient of variation (CV) for total precision was 4,7% at 9,0 mmol/L and 3,7% at 40,0 mmol/L. The CV for within-run was 14,7% at 1 mmol/L, 5,2% at 4 mmol/L and 1,4% at 17 mmol/L. The precision of the method is most important at lower concentrations. The evaluation of linearity showed linear correlation at concentrations <50 mmol/L with. A linear correlation was observed at concentrations >50 mmol/L, although the agreement with the calculated concentrations decreased. The method verification was successful as the results were deemed acceptable for clinical use.

<p>Rapporten laddas upp av lärare om detta godkänns av handledare.</p>

The pre-analytical phase is known for being the most important step in the laboratory process to reach reliable test results. If handling, transport or preparation of the sample is performed incorrectly the results can deviate from the true value. Today, sampling tubes contains various additives to stabilize concentration levels. The aim of this study was to test a new sampling tube containing fluoride/citrate for glucose, lactate and homocysteine. It was also of interest to evaluate the stability of those three analytes in lithium-heparin, sodium-fluoride/potassium oxalate and fluoride/citrate tubes. To perform the sustainability study, a method verification was done for homocysteine in plasma. The study was performed in a hospital laboratory on the routine instrument Roche Cobas 6000 analyzer. Blood was drawn from 20 patients and was analyzed at the hospital laboratory in Gävle. The blood samples were transported frozen to the laboratory in Hudiksvall and were used in the method verification. For the sustainability study, blood was drawn from 10 healthy volunteers in lithium-heparin, sodium-fluoride/potassium oxalate and fluoride/citrate tubes. The method verification was approved. The results showed that glucose was stable for up to 72 hours in Vacuette Glycaemia tube with fluoride/citrate and this tube also gave more accurate results. Lactate and homocysteine were also stable in fluoride/citrate, but needs further studies. All three analytes were more stable if the sample tubes were centrifuged as soon as possible after blood collection. Fluoride/citrate tubes were stable without centrifugation directly.