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AZ Damiaan Oostende

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Abstract Bachelor Project MLT 2020-2021: Validation of a new kit to detect a Mycoplasma pneumoniae infection, serology

AZ Damian Ostend wants to use new ELISA kits for the detection of Mycoplasma pneumoniae antibodies. This research compares the results of new EUROIMMUN kits with the known results of their current kits of Labsystems Diagnostics. There are no clear-cut symptoms that are specific for a M. pneumoniae infection which makes diagnosing very difficult. It is important to detect an infection because there are specific antibiotics for treatment. The aim of this research is a validation of the new kit to find out if the new kit is as efficient as their current kit.

There were 46 samples previously analyzed with the old kits, that were now analyzed with the new kits. These results were then compared to each other. The experiment also tested the specificity of the kits by analyzing different samples, that were positive for other infections, with the new IgM kit. Finally, the sensitivity was tested with a PCR of nasal swabs of several patients. In each run, there were controls that were tracked and compared with the other runs by calculating the mean, standard deviation, coefficient of variation, bias and the total error. Fifteen out of 46 showed discordant results in comparison to the known results of the current kit. The discordant results were false negative or false positive due to the age of the patients, cross-reactivity or the insensitivity of the new kit. The sensitivity of the IgM kit is 73,33 % and of the IgG kit 85,19 %. The specificity of the IgM kit is 90,32 % and of the IgG kits 100,00 %. The result of the PCR test is that all the samples tested negative for Mycoplasma pneumoniae DNA.

The lab has some demands for the tracked controls. The total error has to be less than 50 % and the coefficient of variation may be maximum a third of the total error, around 16,67 %. The control of IgM has a total error of 2,18 % and IgG 8,42 %. The coefficient of variation of the IgM control is 0,92 % and of the IgG control is 3,43 %.

The lab chooses to use the kits of Labsystems Diagnostics, because of the bad cross-reactivity with the EUROIMMUN kit. The kit of EUROIMMUN showed three cases of cross-reactivity and the kit of Labsystems Diagnostics none.


Abstract Bachelor Project MLT 2019-2020: Legionella pneumophila antigeen detectie op urine

Legionella pneumophila is a bacteria that can cause serious diseases in humans such as the Veteran disease and the Pontiac fever. It is therefore important to diagnose this bacteria quickly before irreversible damage occurs to the patient. It is not possible yet to determine the bacteria through urine antigen-detection in the AZ Damiaan lab. The aim of this study is therefore to compare three kits for the determination of Legionella pneumophila in urine. The kit that scores best will then be routinely used by the lab.

The tested kits were BinaxNow (Abbott), ImmuView (International Medical) and CerTest (Selinion). The three kits were tested for specificity, sensitivity, PPV, NPV and accuracy. This was done using 20 positive samples. The kits were also compared based on the price-quality ratio, inclusion of the controls, ease of use, logistics and distribution.

The BinaxNow kit and the ImmuView kit both achieved high sensitivity and high specificity, while the Certest kit achieved a high sensitivity, but a low specificity. The Certest kit is the cheapest, but if the other criteria are compared, the BinaxNow kit comes out best. The kit uses a device to read samples, is easy to use, includes a positive and negative control as standard and the company already supplies other kits to the lab.

The Abbott BinaxNow kit scored best on both statisticall and on other criteria tested. This kit will therefore be included in the routine of the lab

Abstract Bachelor Project MLT 2018-2019: Analytical and clinical evaluation of ‘Elecsys Active B12’ on the Cobas 6000 Device
Vitamin B12 deficiency is common in mixed patient populations and can cause megaloblastic anemia and irreversible neurological problems. However, there is no single marker which can reliably diagnose B12 deficiency. Furthermore, controversy exists regarding the best diagnostic approach. The aim of this study is to evaluate whether active B12 is a more reliable indicator for vitamin B12 status than total B12. First, this study evaluates the analytical characteristics of the kit ‘Elecsys active B12’. It is found that the intrarun precision, the interrun precision, the bias, the total error and the linearity of the aforementioned kit meets the specified criteria of Ricos and Sciensano. Second, this study investigates the clinical usefulness of active B12 as a first- line screening procedure for vitamin B12 deficiency. Active and total vitamin B12 were measured in eighty-eight serum samples (Cobas 6000; Roche). In the grey zone of active B12 (25-75 pmol/l) and/or total B12 (150-400 ng/l), MMA was tested by LC-MS as a reference marker (> 40,1 µg/l). Critical evaluation of the optimal cut-off values in the grey zones indicates that active B12 (cut-off 50 pmol/l) is not more sensitive than total B12 (cut-off 245 ng/l) (59 % versus 59 %), but the specificity is higher (71 % versus 44 %). Thus, it can be concluded that active B12 has a higher specificity than vitamin B12. The use of active B12 for B12 deficiency, however, is not reliably enough as single marker. Hence, the recommended procedure for testing for vitamin B12 deficiency should start with an active B12 measurement followed by MMA testing in the grey zone. This diagnostic two-step algorithm has the best sensitivity and specificity but is still too expensive to implement.
Abstract bachelorproef 2016-2017Comparison of two ELISA kits for the detection of Varicella zoster virus antibodies in serum

AZ Damiaan Oostende wants to perform the detection of Varicella zoster virus antibodies in-house on the DS2 platform from Dynex®. The aim of this validation is to evaluate two ELISA kits from different companies (EuroImmun and Vircell).

The detection of Varicella zoster virus antibodies is important for several patients. Pregnant women who get in contact with the virus, need to know their immune status for the protection of their unborn child. There are several kits available for detection.  

Different samples were validated and compared on both kits in a short time-frame. Several runs were performed on the DS2 platform with the same samples under the same conditions to compare the results of both kits. The samples were tested for both IgM- and IgG-positivity and compared to the results obtained by the laboratory AML, where these tests were done on a chemiluminescence immunoassay platform. The controls of each run were tracked and compared to each other by calculating mean, standard deviation and coefficient of variation. Specific samples were tested to see if there was any cross-reactivity to Epstein-Barr virus, Herpes simplex virus, Reumafactor and Cytomegalovirus in the detection of IgM-antibodies.

The coefficients of variation were for both IgM and IgG from both kits less than 10%. There was only one sample out of 28 that was discordant for.

The IgM kit from Vircell had significant more cross-reactivity than EuroImmun.

EuroImmun IgM, IgG and Vircell IgM and IgG showed a sensitivity of 100%. The specificity was 100% for EuroImmun IgM, Vircell IgM and IgG. The kit EuroImmun IgG had a specificity of 83,3%.

Due to the lower cross-reactivity and practical considerations, the laboratory chooses to use the IgM/IgG-kits from EuroImmun.

Samenvatting eindwerk 2012-2013: Bloedtransfusie: een eerste stap richting Type & Screen
Een bloedtransfusie van erythrocyten kan enkel worden uitgevoerd na de bepaling van de bloedgroep van de patiënt en de compatibiliteit tussen donor en receptor via ‘Crossmatch’ of ‘Type & Screen’. Het doel van deze bachelorproef was het evalueren van het bloedtransfusie-beleid, namelijk ‘Crossmatch’ versus ‘Type & Screen’ en het vergelijken van de twee grote spelers op de In Vitro Diagnostics markt, namelijk DiaMed versus Ortho Clinical Diagnostics. In het laboratorium van het AZ Damiaan te Oostende worden alle testen binnen de immunohematologie uitgevoerd op de typeerkaarten van DiaMed, waarbij de compatibiliteit wordt nagegaan via klassieke kruisproeven.
Om te voorkomen dat getransfundeerde donor-erythrocyten versneld afgebroken worden, met als gevolg transfusiereacties, is het belangrijk dat een patiënt geen donorbloed ontvangt waar hij antistoffen tegen heeft. Om de compatibiliteit tussen donor en receptor te garanderen worden compatibiliteitstesten uitgevoerd. Bij ‘Crossmatch’ wordt contact gemaakt tussen het serum of plasma van de receptor en de erythrocyten van de donor. ‘Type & Screen’ daarentegen garandeert de compatibiliteit op basis van een screening naar irreguliere antistoffen en een elektronische kruisproef. De aan- of afwezigheid van irreguliere antistoffen wordt nagegaan door het uitvoeren van een Indirecte Coombs.
De typeerkaarten van DiaMed en Ortho Clinical Diagnostics zijn sterk vergelijkbaar. Ze verschillen enkel in het medium dat gebruikt wordt om een onderscheid te maken tussen geagglutineerde- en niet geagglutineerde erythrocyten.
Om een evaluatie van beide luiken mogelijk te maken, werd bij elke aangevraagde kruisproef een bloedgroepbepaling en Indirecte Coombs uitgevoerd volgens zowel het systeem van DiaMed, als van Ortho Clinical Diagnostics. In geval van een positieve Indirecte Coombs werd er eveneens een identificatie van irreguliere antistoffen uitgevoerd.
In het AZ Damiaan worden zo’n 40% van de aangevraagde units packed cells niet toegediend. ‘Type & Screen’ zou dus zinvol zijn om op te starten, en voornamelijk op de chirurgische diensten. Indien gewerkt wordt volgens ‘Type & Screen’ wordt er namelijk geen contact gemaakt tussen donor en receptor. De units packed cells worden als dusdanig niet voorbehouden voor een bepaald persoon en kunnen op elk moment worden vrijgegeven. Één Indirecte Coombs in 72u volstaat bij ‘Type & Screen’ voor het uitboeken van meerdere units packed cells, naast telkens een elektronische kruisproef. Deze methode zou dus tijdswinst en een optimaler beheer van de bloedbankvoorraad kunnen opleveren. De aanwezigheid van laagfrequente antigenen wordt echter, in tegenstelling tot de klassieke kruisproef, niet uitgesloten. Wegens de mogelijkheid tot verdere automatisatie kan de personeelskost dalen, wordt de kans op menselijke fouten steeds kleiner en is er verdere standaardisatie van het bloedtransfusie-beleid mogelijk. Bovenstaande waarnemingen en interpretaties kunnen pas bevestigd worden na de uitvoering van de methode ’Type & Screen’ in de praktijk.    
Beide spelers, binnen de In Vitro Diagnostics markt, vertonen zowel voor- als nadelen. Kiest men voor het behoud van de huidige methode, dan neigt de voorkeur, wegens de gemakkelijkere manuele aflezing, naar DiaMed. Overweegt men de overstap naar ‘Type & Screen’ aan de hand van een bloedtransfusie-automaat, dan neigt de voorkeur, wegens de meer gevoelige Indirecte Coombs, naar Ortho Clinical Diagnostics. De keuze tussen beide systemen op basis van de bloedtransfusie-automaten werd in deze bachelorproef niet behandeld en is voer voor eventueel verder onderzoek. 


Gouwelozestraat 100
8400 Oostende
059 41 40 40


Traineeship supervisor
Apr. L. Vynckier
Lies Persijn
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