BLT Stages

Abstract Bachelor Project MLT 2020-2021:  Sensitivity determination of anaerobic germs using Micronaut-S microdilution

We evaluated the performance of the MICRONAUT-S microdilution broth assay, relative to the Antibiotic gradient test of Lyofilchem™, test strip by comparing the minimal inhibitory concentration (MIC) of different anaerobic bacteria against  different antibiotics. At present, according to European Society of Clinical Microbiology and Infectious diseases (EUCAST), there is no reference method for the antibiotic susceptibility assessment of anaerobes. Currently, the most commonly used method, the antibiotic gradient test strip, is laborious, time-consuming, burdensome for interpretation and reproducibility problems, which often hamper routine workflows. The microdilution method could be a great advantage in the routine, in terms of standardization and interpretation of the results. By using both tests simultaneously, the categorical and essential agreement between both methods were assessed. Furthermore, the hand on time, the practical and methodical advantages for both techniques for the lab technicians were determined. When the results of both methods are evaluated against the MIC values from reference isolates, it can be observed that the Micronaut-S showed better agreement than gradient test strip. For this kind of experiment, the repeatability, the reproducibility, the categorical and the essential agreement are above 90 percent (%) as required. When comparing between the two methods and the results versus the known MIC value, the CA is superior with the Micronaut-S than with the gradient strip test. The CA for the Micronaut-S at both ATCC and reference strains is 65.8% and for gradient strip it is 80.7%. Comparison relative to each other is 83.4%. Labor intensity and the hand on time is equal for both, also the purchase price is equivalent. The biggest difference would be in standardization, reading the results and interpretation, for which the Micronaut-S is considerably more straightforward.

 

Patients suffering from certain haematological disorders may undergo a hematopoietic stem cell transplantation. At AZ Sint-Jan, site Bruges, pre- and post-transplant samples are taken and analyzed with a high-throughput sequencing based assay. The resulting data of screening (pre) and follow-up (post) samples are currently analyzed with the ADVYSER software (DEVYSER). This software calculates the host chimerism percentages for follow- up samples based on the sequence data. Drawbacks of the software are the lack of traceability, limited visualization capabilities hampering interpretation efficiency and reliability, and lack of networking possibility (the current access to the software is only possible via one computer in the lab). A new webtool was constructed to solve these disadvantages.

To construct the web-application and visualize the results, several computer languages are used. To construct the GUI a front-end open source toolkit, known as Bootstrap, was used. The HTML code, with styling via Bootstrap, was combined with PHP to upload and process data, and eventually display the results through a web interface. Uploaded data is stored partially in a MySQL database consisting out of two tables (screening and follow-up), and partially in a folder structure. Depending on the sample, data will be written to either the screening table or the follow-up table, and to specific files in the folder structure.

To facilitate interpretation, a preference was to show the different results in a more graphical way, as opposed to the ADVYSER software which provides a HTML text report only. To visualize the results on the webtool, several figures and tables are constructed for each sample type. This was performed via R. Upon file upload, a trigger will activate the R scripts and the figures are constructed.

Taking into account that mistakes may happen when uploading a sample, a function to edit or delete data was implemented. Via a webform the user can easily modify the information of a screening or follow-up sample, or delete an existing sample. To prevent major problems when performing this action, two checkups are implemented. A screening sample can only be modified if no follow-up sample is available. On the other hand, from a set of follow-up samples of a patient, only the most recent one can be modified. These different actions are all performed via HTML and PHP.

For easier navigation and adding more functionality to the web-pages, several Javascript functions were implemented. This was mostly done for showing different alerts and adding lookup tables for easier use of the webtool. According to these available functions, it can be concluded that the most important functions are already available. Yet the webtool is not final. Several aspects still need to be implemented. An example is the integration of an existing log-in system which will improve the traceability.

 

For my internship I was asked to implement a web interface for monitoring quality parameters of Illumina runs. The data of these runs could already be visualized by the free application Illumina sequencing analysis viewer. This application has one major downside, it is not possible to compare values for the quality parameters between the different runs and samples. Being able to perform this on the web interface was the goal.

We worked in R to extract and read all the data for the runs. Using the packages XML and SavR these files could be handled in an R-script. SavR is used to parse and analyse Illumina SAV files, these files contain the quality parameters for one run. In the XML files, general info about the run can be found, such as run number and date. For some parameters extra steps were needed; calculating averages or discarding 0-values for example.

After processing all this data and calculating the needed values, these values are stored in a MySQL database thru the R-script. In the database we make a distinction between runparameters and sampleparameters. For the runs there are 23 different parameters in the database: RunID, Name, Total Reads, PF Reads, PF Percentage, Reads Identified, Q30, Density, PhasingR1, PhasingR2, PhasingR3, PhasingR4, PrephasingR1, PrephasingR2, PrephasingR3, PrephasingR4, Read1Length, Read2Length, Read3Length, Read4Length, Tiles, Date and Instrument. For comparing the samples only 4 parameters are used: RunID, Name, Fraction PF Reads and Reads.

Now that a database is created with all the data, we can call for these values, representing each run, to create plots and make it easy to compare different runs. For each parameter a separate script was created. In general, these scripts call for the info for each run in a loop for the corresponding parameter. After that a plot is made with this data and saved as an image. These images will be used in the web-interface.

For the actual web interface, we worked in html and php(notepad++). The bootstrap framework is used for designing the layout. It includes HTML and CSS based design templates and supports JavaScript plugins. In the php files a lot of interaction happen between the php file itself and the MySQL database.

The web interface contains 4 different start pages. The first one contains all info of the most recent run and comparison with the 20 previous runs. The second one is a list with all run numbers where the most recent one is on top off the page. The third one is the same but for samples. The last page is a list of instruments used between all the runs.

When clicking a run or sample on these pages, you get redirected to a details page for this particular run or sample. On this details page you can find a table with the values for all parameters. Underneath the table, you can find the comparison between the 20 previous samples/runs and the selected sample/run in the form of different graphs. Some graphs have cut-offs where a colour code can be applied. When clicking on an instrument, a list of run numbers is given. All these pages are linked together.

In the end we created a clear and simple web interface for comparing runs and samples. In addition an instrument page was created and the latest run has its own page. The script also runs automatically when a new folder with data is added to the parent folder.

Abstract Bachelor Project 1 MLT 2019-2020:  Validation of the BD BACTEC^TMFX for sterility testing of products from the bone and tissue bank and the hospital pharmacy

The current method of sterility tests at the pharmacy and tissue bank of the AZ Sint-Jan Hospital in Bruges shows a number of shortcomings such as the fact that it’s very labor-intensive. These sterility tests are used to verify the absence of viable and actively multiplying micro-organisms in preparations. This is done to prevent intravenous infections.

The plan is to replace the current manual method with an automated method using the BACTEC^TMFX Blood Culture System.

In this research, different pharmacy preparations (smoflipid/vitalipid, norepinephrine, growth 92 without trace elements and PCA epidural), stem cells and bone product are spiked with different types of micro-organisms. These suspensions are injected into a hemoculture bottle with a sterile syringe and needle. The hemoculture bottle is incubated in the BACTEC system until it’s flagged as positive which means that it can adequately detect growth. After this, micro-organisms are identified in the positive bottles. This identification is done with microscopy (Gram stain), culture and massaspectrometry (MALDI-tof).

In the first project, the bone and tissue bank project, the BACTEC method is compared to the standard plating method. This comparison led to the conclusion that the BACTEC method is faster than the plating method (2.56 vs 11.18 days). The BACTEC system is also the most sensitive and therefore the most accurate to identify C.albicans (53.85 vs 15.38%), as well as A.fumigatus (46.15 vs 38.46%) and Fusarium spp. (38.46 vs 0.00%). For the second project, with pharmacy preparations, the BACTEC system was validated in this study. The BACTEC method was found to be a sensitive method for different types of micro-organisms: S.aureus (100.00%), B.cereus (87.50%), P.aeruginosa (100.00%), C.albicans (87.50%) and A.fumigatus (75.00%). However it appeared to be less sensitive for anaerobes (C.perfringens 43.75% and B.fragilis 50.00%). This is not due to the BACTEC system but is caused by coming into contact with oxygen when making dilutions.

It can be concluded that the BACTEC system is a much easier and faster method with less hands on time, than the classic plating method when it’s used with suspensions that include bone product or stem cells. The method is very sensitive with pharmacy preparations as well. The current manual method can therefore be replaced by the automatic BACTEC method in the pharmacy and tissue bank of the AZ Sint-Jan Hospital in Bruges.

 

Abstract Bachelor Project 2 MLT 2019-2020:  Evaluation of automated tissue dissociation for flow cytometric analyses

The purpose of this research is to evaluate a new device (the MediMachine II) for automated tissue dissociation for flow cytometric analyses.

The current method for automated tissue dissociation is the GentleMacs Dissociator. Although already in use for several years, two main problems do occur. Sometimes an extra population is seen after an analysis, but there is no explanation for which cells this population indicates. The second problem is that sometimes malignant populations cannot be found with flow cytometric analysis. For these reasons, an alternative device will be evaluated (the MediMachine II). This device could be a solution for avoiding the two problems with the GentleMacs Dissociator.

To evaluate the device some parameters are examined. These are the correlation and difference in viability. These parameters will also be carried out with the manual method because it is known that with this method the least cells are destroyed. Also, the imprecision and the shelf life of a sample will be examined an as additional tests a price comparison will be made and the difference in ease of use will be examined. These tests will be done on lymph nodes and will be analyzed with the BD FacsCanto II flowcytometer.

The results of the tests show that the MediMachine II had higher correlation results for the analysis of CD19+. This can be important for analyzing B-cell chronic lymphoproliferative diseases. The difference in viability of the cells between the devices is notable. The stopping-gate of 5000 lymphocytes is more achieved with the MediMachine II than with the GentleMacs Dissociator. This may be due to the mechanical processing with the GentleMacs Dissociator, causing more destroyed cells. The imprecision is good with only a few values outside the limits. This is due to the heterogenicity of the tissue. Analyzing a lymph node is best within 24 hours. After that, the results can no longer be considered reliable. From the previous tests the MediMachine II is better but looking at the price and the ease of use, the GentleMacs Dissociator is better. The MediMachine II is more expensive and requires a manual and automatic processing step.

The results of the evaluation show that the viability of the cells is better with the MediMachine II and the manual method. This may be the reason that maligned populations with the GentleMacs Dissociator are more difficult to perceive. This parameter is an advantage for the MediMachine II. The switch to the MediMachine II will also depend on the other parameters because it’s more expensive and less easy to use.

 

Abstract Bachelor Project 3 MLT 2019-2020:  Evaluation of the use of IHC to detect MMR deficiency / micro satellite unstable tumors, in particular colorectal carcinomas

Lynch syndrome is an autosomal hereditary disorder that is the result of a mutation in one of the MMR-proteins (MLH1, PMS2, MSH2 and MSH6). Patients with Lynch syndrome have 25 to 70% chance to developing colon cancer. The aim of this research is to evaluate the correlation between micro satellite instability (MSI) and immunohistochemistry (IHC). There is also an evaluation of the reliability of a two- antibody panel.

MMR deficiency can be detected by immunohistochemical staining and / or MSI analysis. This study is about immunohistochemical staining. These stains are performed on the MSI panel of the Roche Ventana device. The indirect detection method is the method most frequently used on the Ventana.

There is a 98,37% correlation between MSI and IHC. There is 0,00% possibility of false positive results and a 1,63% possibility of false negative results. When there are ‘possible’ results, these tissue sections should be checked for any misinterpretation of the IHC because these tissue sections show abnormal expression. The Shia et al study confirmed that a two-antibody panel is as reliable as a four-antibody panel. Isolated loss of PMS2 and MSH6 gives a greater indication of Lynch syndrome than MLH1 and MSH2.

There is a good correlation between MSI and IHC. Both techniques will continue to be applied at AZ Sint-Jan in Bruges. From this study there can be concluded that the IHC is reliable if an MSI analysis confirms colorectal carcinomas. The four-antibody panel is still used. In the future, a two-antibody panel can be introduced in the lab.

 

Punching was performed from FFPE tissue from oropharyngeal tumors. The DNA was extracted using the MagCore, then diluted and placed on the Cobas. The results were compared with those obtained from the p16 IHC and HPV ISH, tested on the Benchmark Ultra from Ventana (Roche). Two methods for measuring DNA concentration were compared, showing that the Qubit is more reliable than the NanoDrop. By comparing different diluents, it was decided that the extract should be diluted in 1 ml of preservcyt or 1 ml of 50% ethanol. It was tested on two samples whether the extraction step with the MagCore could be omitted and could be extracted manually instead, but this must be further tested with more samples. The quality of the DNA was tested on the basis of a QuantiMIZE run. This test showed that the DNA from the oropharynx was damaged or too fragmented. Discrepant results were obtained for two of the ten HPV HR+ samples. Two of the ten HPV HR+ samples gave an invalid value. Some of the other 6 samples were positive for HPR16, but invalid for other HR HPV and HPV18. Invalid results were all obtained for the four oropharynx LR+ samples. The invalid values can be explained by the fragmentation of the DNA.

 

Abstract Bachelor Project 2 MLT 2018-2019: Reducing the turnaround time of methicillin-resistant Staphylococcus aureus screening

Abstract Bachelor Project 3 MLT 2018-2019: Screening of organ-specific autoantibodies in liver diseases: a comparison of three liver/stomach/kidney substrates from the rat and the mouse with classical versus automated fluorescence microscopy

 

In Belgium, until recently, there was no uniform, agreed-upon algorithm to determine the biological significance of a variant. In the Molecular Hematology lab at AZ Sint-Jan BruggeOostende, the significance of a variant is currently determined by a combination of 6 parameters: frequency in the COSMIC database, not frequently occurring in dbSNP/gnomAD/1000Genomes, SIFT classification, Polyphen classification, variant allele frequency (VAF) and whether or not the variant is located in a sequence encoding a conserved domain. Recently, a national work group has agreed upon an algorithm for the biological classification of variants. Based on these new rules, the biological class of a variant will be determined by the following parameters: frequency in the COSMIC database, classification as determined by SIFT, MutationTaster, JAX CKB, MDA, MCG, CIViCdb, ClinVar, OncoKB and Varsome, gnomAD frequency and evidence as available through PubMed. Each variant can be classified using this data according to a scoring algorithm. The goal of this is to make the process uniform across all hospitals in Belgium, so that the biological significance of a variant is the same in every hospital. The goal of the traineeship was to build a platform for storage, management and analysis of variants detected by high-throughput sequencing and the additional parameters required for biological classification. The three main pillars of this platform are 1. a MySQL database to store the variant data generated by the Illumina MiSeq, along with required parameters and metadata, 2. R scripts to parse the variant data files and annotate the variants in these files using information retrieved from online and offline databases, 3. a dynamic web tool written in HTML/CSS/JS/PHP which queries the MySQL database.

 

Consequently, the blueprint of the database was determined by these new requirements. The main database consists of 7 tables: (i) a panel table which has the basic data of the different panels of genes; (ii) a run table which contains all the particulars from each time an experiment is run; (iii) a sample table which incorporates the necessary information of all the samples within the runs; (iv) a patient table which holds the patient details of the samples; (v) a variant table which stores all the variants that are detected by highthroughput sequencing; (vi) a variant-sample table which functions as a cross reference table between the sample and variant table; (vii) and finally a version table which shows when the annotation data of the linked databases was last updated. Additionally, there are 7 trace tables which log all the changes that are made to the 7 main tables, one for each, based on MySQL triggers. This is to ensure traceability of both automated and manual changes. Lastly, there are three tables for the ClinVar data: a ClinVar table with all the ClinVar information considered relevant, a PubMed table with the PubMed documentation associated with a variant and a ClinVar-PubMed table which functions as a cross-reference table between the former two.

 

Using R scripts, the variant data files can be parsed and the important information can be extracted and inserted into the database using RMySQL. Other R scripts are then sourced to update the detected variants with additional annotation data from the genomic and other databases listed above. At this time, it is completed for COSMIC, dbSNP, SIFT, Polyphen, JAX CKB, ClinVar, gnomAD and PubMed. The majority of this information is retrieved through the Ensembl Variant Effect Predictor API. The NCBI Eutils API is used to retrieve ClinVar and PubMed data. gnomAD and JAX CKB data are downloaded and subsequently parsed. In the past, all the important variant data was manually curated in an Excel file. This old data also had to be inserted into the database. A separate R parsing script was created to extract all the useful information from the Excel table.

 

The web tool allows for the management and analysis of the variants stored in the database. It is mainly written in HTML, CSS and PHP, but also has some JavaScript for specific tasks. MySQLi prepared statements are used inside PHP to query the database. This results in a set of interconnected pages featuring the data from all the tables in the database which can be navigated through the navigation bar and the action buttons. The styling of the interface is created using Bootstrap.

 

The benefit of this endeavor is that the information of the variant data files is now automatically parsed and stored in a database instead of it being manually inserted by a lab technician or a clinical biologist into an Excel table. This saves time and reduces mistakes due to manual curation, the storage system is more reliable and allows for tracing changes, and the web interface makes it easier to manage and analyze the variant data. 

 

Abstract bachelorproef 2017-2018: Co-testing of high risk human papillomavirus with the Cobas 4800 instrument

Cervical Cancer is one of the most common cancers in Belgium with 1414 new cases detected in 2011. The cancer is associated with the human papillomavirus (HPV) which is sexually transmitted. The traditional detection of cervical cancer is only based on the morphological image of the cells, but if the virus is latently present in the cells then is a morphological image unable to detect the virus. The advantages are an early detection of an HPV infection to start an early treatment, a better follow-up for the patient.

The aim of this thesis is to show the importance of co-testing. Co-testing is making a smear and colored with a Papanicoloau staining next to an HPV-testing on a sample with the Cobas 4800 from Roche independently of the result of the Papanicoloau staining.

If the smear is negative, the woman will be adviced to making every three years a new smear. Co-testing will also test for the HPV-DNA and when the sample contains HPV-DNA, the woman will be adviced to come back after one year for follow-up.

In this first part of the study is a global image about human papillomavirus, how works the traditional screening and what is co-testing.

In this second part of the study is tested the precision, reproducibility and a comparison of the positive NILM-samples between the Cobas 4800 and the validated MTA method.

The results in the study showed that 9,7% from the negative samples in the screening has a HPV-infection. There are 468 samples tested independent of the results of the screening (NILM, LSIL and HSIL). There is a 100% sensitivity and specificity with samples of the external quality control. Also, are there 93% of the HSIL samples positive for HPV tested with the Cobas 4800.

The Cobas 4800 is a good method for co-testing. When the comparison is tested between the Cobas and the MTA is the Cobas more sensitive for detect lower concentrations of HPV-DNA. The correctness is also good and is tested with external quality controls and with HSIL-samples.

 

Myelodysplastic syndrome (MDS) are a group of clonal haematopoietic stem cell diseases characterized by peripheral cytopenia(s) and dysplastic morphology. The diagnosis relies on bone marrow (BM) cytomorphology and cytogenetics. However, recognition of dysplasia in BM smears can be challenging and cytogenetic abnormalities are not always found. On the other hand, the dysplastic features and cytogenetic abnormalities can also be found in conditions other than MDS and in healthy persons. So the diagnosis of MDS is not always straightforward. This thesis aims to evaluate the value of flow cytometry in the diagnosis of MDS.

First there was an evaluation of a flow cytometric score, the Ogata score. The four parameters used in this score are: myeloblasts(%), B-cell progenitors (%), myeloblast CD45 expression, and side scatter of granulocytes. These parameters can be determined with the Lymphoid Screening Tube (LST) that is analysed in every bone marrow sample of a patient without a history of a haematological disorder in AZ Sint-Jan. So, the Ogata score can be determined without an extra cost.

The second method was an extended Ogata score with CD56 expression on the myeloblasts and the monocytes (%). This parameters can also be determined with the LST.

For these methods a total of 37 MDS samples (22 low risk MDS and 15 high risk MDS) and 76 control patients with cytopenia were retrospectively analyzed.

For the third method a new MDS-panel was designed based on literature, in this panel the following parameters were used: CD45, CD15, CD56, CD34, CD11b, CD10, CD19 and CD7 each with a different function. For this prospective evaluation four MDS samples and four control samples were used.

The sensitivity of the Ogata score was 65% and the specificity 92%. For the extended Ogata score a sensitivity of 81% and specificity of 54% was found but their was a problem with an aspecific connection between CD56 and kappa on fluorochroom PE, result in false positive results. With the new MDS panel there wasn’t an added value with the Ogata score to distinguish MDS cases from control cases.

Flow cytometry can be used based on the standard Ogata score as a diagnostic tool for MDS. Because of the high specificity, the diagnosis can be suggested when a positive score is found. Moreover, the analysis can be performed without an extra cost.

Abstract bachelorproef 2 2016-2017: Validation Optilite® CH50 Reagent on Optilite with reference method Wako® Autokit CH50 on P-Modular

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Abstract bachelorproef 3 2016-2017: Is the Idylla device of Biocartis effective for routine use in detecting EGFR / KRAS / BRAF mutations?

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Abstract bachelorproef 1 2015-2016: Evaluation and comparison of four different analyzers for the quantification of HbA1c

To evaluate and compare four commercially available HbA1c analyzers (Menarini HA-8180, Bio-Rad D-100, Tosoh G8 and Sebia Capillarys 2 flex piercing) with respect to (1) correlation against the current laboratory HbA1c analyzer (Menarini HA-8160) and (2) imprecision against the HbA1c analytical goal of the coefficient of variation ≤ 2.9%.

Glycated hemoglobin (HbA1c) has become the preferred method for monitoring diabetes over the last decade. Various commercially available analyzers have been developed for measuring HbA1c in a routine laboratory. The analyzers should deliver fast and accurate results providing both the practitioner and the patient with the information for an appropriate diagnosis, treatment and follow-up of the patient.

The validation and comparison of the four analyzers are performed over a period of 2 weeks. During this period a series of standardized tests are executed and operator observations are recorded. The accuracy of each analyzer, including the laboratory analyzer, is assessed by using the calibrated SKML controls for both a low and a high HbA1c sample. The maximum allowed mean bias is 2 mmol/mol. The repeatability is performed by the consecutive measurement of ten low and ten high samples on each analyzer, whereas the reproducibility is executed by measuring ten identical both low and high samples twice a day during five consecutive days. For both criteria the calculated coefficient of variation CV must be maximum 2.9 %. The linearity is obtained by measuring test samples with linear increasing concentrations of 30 to 80 mmol/mol. A minimum R² of 0.95 is considered to pass this test. In order to verify the correlation of the new analyzers compared to the current laboratory analyzer (Menarini HA-8160) a series of 128 samples, including five hemoglobinopathy samples, is measured. Calculations and graphs of the linear regression with correlation coefficient r, the Intercept A, slope B at a confidence level of 95%, are performed using Passing Bablok regression and Bland Altman plot. Carry-over tests to check how efficiently the analyzer can manage eventual contamination by a previous high HbA1c sample on the next normal samples are executed. Lastly the functionality and the observations made while testing, in particular the features, the complexity, the required preparations, the quality of the reports, the software and the user interface are evaluated.

The coefficient of variation is acceptable for all four analyzers, although the CVs of the repeatability and the reproducibility for both the Menarini HA-8180 and the Tosoh G8 analyzers are significantly lower than for the two other instruments. In respect to linearity only Tosoh G8 (R² = 0.947) is slightly below the target. The Capillarys of Sebia shows the best linearity (R² = 0.9988). Passing-Bablok regression executed on the four analyzers compared with the current Menarini HA-8160 shows only for Sebia an intercept A outside the confidence interval of 95%. Carry-over does not appear on either analyzer.

The analyzer with the best user interface, the least complexity and generating the fastest and the best reports is the Bio-Rad D-100 analyzer.  

The Menarini HA-8180 is the best choice considering all criteria with acceptable precision, good linearity and acceptable agreement with the central laboratory results. The more only this instrument provides the feature of obtaining the HbA2 concentration simultaneously with the HbA1c measurement. Also in regard to simplicity of operation, fast and reliable reporting, the Menarini HA-8180 scores high.

 

In the AZ Sint-Jan Hospital Campus Brugge an HPLC system (Menarini HA-8160) is used for analysis of different types of hemoglobin: HbA1c, HbA0, HbA2, HbF and hemoglobin variants. With this system different clinical manifestations can be detected such as monitoring of patients with diabetes by the quantification of HbA1c and detection of hemoglobinopathies (thalassemias and hemoglobin variants) by quantification of HbA2, HbF and any hemoglobin variants. After thirteen years of duty, this system needs to be replaced, therefore an evaluation of three new HPLC systems and one capillary electrophoresis is performed on demo systems to decide what system would replace the currently used one. In this thesis the evaluation of the four systems for analysis of hemoglobinopathies is displayed. The three HPLC systems are Menarini HA8180T, TOSOH G8 and Biorad D-100. The capillary electrophoresis system is Sebia Capillarys. These where tested on quality control, between run, intra run, correlation, linearity, carry-over and advantages and pitfalls for using them. During these tests 64 blood samples where used, these contained samples with α-and β-thalassemia’s, and hemoglobin variants such as HbA/S, HbS/S, HbA/E, HbA/D-punjab, HbA/C, HbA/Lepore, HbA/J-baltimore and HbA/Muravera. The results showed that every system had its pitfalls and its advantages, although there were no major deviations with clinical significance. This means that in the end every system has successfully passed the evaluation.

Worldwide there were more than 500.000 new diagnoses of cervical cancer in 2012. With a mortality of about 50%, 250.000 women die of this disease annually. Over 99% of these cancers are caused by high risk human papilloma viruses. The quicker the treatment, the better the prognosis. Therefore, the early detection of the virus is very important.
The main purpose of this study is to compare the Cepheid Xpert HPV-assay and the Hologic Cervista HPV-HR test. Both methods test for high risk HPV on cervical PAP-smears. The comparison include hands-on time, runtime, verification and workload.
The Cepheid company has provided 30 HPV high risk screening tests. From our recent files 30 PAP-samples were selected upon which Cervista HPV high risk tests were already performed. The 30 samples include five ASC-US, five NILM, five LSIL and five HSIL-samples. The reproducibility will be tested by testing a proven HPV-positive sample three times on the Cervista MTA and six times on the GeneXpert. The different modules of the GeneXpert will be tested separately.
The HPV-typing of both methods have about the same result. The MTA has a higher sensitivity, sometimes generating false positive results. Because the GeneXpert is single-sample based, the results are immediately available, but generate a higher cost per test. The choice between the two methods depend on the amount of samples that need to be tested. The more samples, the better the MTA is for a technique. In case of only a few samples a day the GeneXpert is preferred, but with a higher cost.