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AZ Sint Lucas Brugge

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Abstract Bachelor Project 2017-2018: VALIDATION OF THE TRANSITION FROM THE CROSSMATCH METHOD TO THE TYPE AND SCREEN METHOD IN A MEDIUM-SIZED HOSPITAL

In AZ Sint-Lucas Bruges they currently use the crossmatch method to determine if the patient has irregular antibodies before a blood transfusion is performed. This method is very labor-intensive and causes stress to the laboratory technicians on busy moments.

The goal of this bachelor’s thesis is to validate the type and screen method in order to replace the crossmatch method, which is possible according to the American Association of Blood Banks (AABB).

However, in order to implement this method in the daily routine of the hospital, the safety needs to be evaluated.

The validation of this method is performed over a period of one month and three weeks. First was checked if the patient was suitable for type and screen. If the patient was not suitable for type and screen, the procedure of the crossmatch method was followed. If the patient was suitable, both the crossmatch and a screening for irregular antibodies were performed and the results were compared.

It can be concluded that the type and screen method is a safer method than the crossmatch method: irregular antibody screening was positive when the crossmatch was positive and five more cases of irregular antibodies were detected.

The implementation of this method can reduce the crossmatches by 62, 05 % in the current setting, which can relieve the workload on the laboratory technicians.

Although type and screen is more expensive than the crossmatch method, there are more advantages than disadvantages and therefore the hospital will implement this method in its daily routine.

 

Abstract bachelorproef 2016-2017Validation of chromogenic medium in screening for Staphylococcus aureus

In AZ Saint-Lucas Bruges, there is a preoperative screening for Staphylococcus aureus to prevent postoperative wound infections. This screening is carried out on all the dialysis patients and orthopedic patients who will have a prostheses. Also the patients of the breast clinic are screened.

They take a nose and throat sample with a TSB salt wiper. Because of the high salt level this is a selective medium for Staphylococcus aureus.

A comparison was made between the CNA-blood agar and the brilliance staph 24 side of the new brilliance staph 24/brilliance MRSA 2 biplate. This is a selective medium for the isolation of Coagulase positive Staphylococci within 24 hours. The CNA-blood agar has a sensitivity of 88.89 % but low specificity and positive predictive value. The biplate has a sensitivity of 100 % and high specificity and positive predictive value which makes the biplate a better screening method for Staphylococcus aureus.

The identification of the colonies is carried out with the MaldiTof. If it is a Staphylococcus aureus an antimicrobial susceptibility testing is performed to check the resistance and sensitivities for the decolonization schedule. 

Abstract bachelorproef 2015-2016Comparative research between RAPIDEC® CARBA NP and KPC/OXA-48 K-Set for the detection of Carbapenemase-producing Enterobacteria

Preventing the occurrence of Carbapenemase-producing Enterobacteria (CPE) is still a major concern for public health in the world, because of the few remaining options for therapeutically treating infected patients. The rapid spread of CPE leads to nosocomial outbreaks and endemic situations in several European countries and all over the world.  Therefore a fast detection and confirmation technique, with an influence on optimal hospital hygiene, is needed.

At A.Z. Sint-Lucas the detection of CPE carriage takes four days using the current method. The first day a wiper, taken from the patient's rectum, is submerged in McConkey broth. It's a selective medium that stimulates the grow of Enterobactericeae. After an overnight incubation at 37 °C, a drop from the McConkey broth is grafted onto a Brilliance CRE medium. After a second overnight incubation at 37 °C the plate is read. The CRE medium is selective, as a certain concentration of a carbapenem antibiotics mixture is added to the medium along with chromogenic substrates, which makes it possible to distinguish between CPE and non-CPE germs. When suspicious CPE stems are present, the E. coli will colour pale pink and KESC (Klebsiella, Enterobacter, Serratia and Citrobacter) will colour dark blue. To confirm the nature of the germ, it is identified through MaldiTOF. Antimicrobial susceptibility testing is performed on Microscan. Where there is a suspicion of CPE, the synergietest and E-test temocilline will be performed. The result will take 4 days. During this traineeship I was asked to examine several rapid testing methods, such as KPC plus OXA-48 K-Set and RAPIDEC® CARBA NP, to determine which are reliable and which can be used in the future to confirm CPE suspected isolates.

There were 35 suspected routine samples and 38 frozen suspected samples examined during the traineeship at A.Z. sint-Lucas, A.Z. Sint-Jan and A.Z. Alma (Eeklo).  Sensitivity and specificity were only calculated from the frozen samples, because only one positive sample was detected in the routine samples. Sensitivity and specificity are two valuable standards by which a rapid test is evaluated. During this research traineeship it was apparent that both the current method and the new rapid test score highly on sensitivity and specificity. The rapid tests KPC/OXA-48 K-Set score the highest on sensitivity and specificity, but are not able to detect all carbapenemasen, which is a hugh disadvantage in the confirmation of CPE.

After putting side by side all results, the current method remains the best method to detect all kinds of suspect carbapenemase germs. The main disadvantage remains the longer delay to get results. In pursuit of patient care, which asks for a quick detection technique, RAPIDEC® CARBA NP offers the best progress for optimum hospital hygiene. When detecting a suspicous germ for CPE the best option is to use the RAPIDEC® CARBA NP test and get a reliable, both specific and sensitive, result. When the test strip gives a positive result, the current method is started for confirmation. By using both, the lower specificity of RAPIDEC® CARBA NP and the lower sensitivity of the current method are avoided. The two results combined give a reliable result.

Samenvatting eindwerk 2014-2015: Schimmelidentificatie met behulp van de Maldi biotyper
In the microbiology laboratory of AZ Sint-Lucas the identification of molds is classically based on macroscopic and microscopic examination. These methods have some disadvantages: it’s a slow process, there’s a need of an expert in mycology and identification to species level is not always possible.
The molds that are being analyzed in the lab are most of the time dermatophytes. These fungi cause infections of the hair, skin and nails. The molds can be analyzed by matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS). This is a technique that has often been used in microbiology laboratories to identify bacteria. The goal of this thesis is to confirm whether MALDI-TOF MS can identify fungi with reliable results.
MALDI-TOF MS analyses proteins and gives a spectrum that can be compared to a database.
Three identification methods will be compared to see which one gives a better result on reliability. The first method is the direct transfer of the mold on the Maldi target. The second method is a short extraction of the mold with formic acid. The last method is a long protein extraction.
The results of these three methods will be compared to the microscopic identification, to see if the Maldi biotyper gives a better result on reliability. If this is the case, the Maldi biotyper can be implemented in the routine of the microbiology lab of AZ Sint-Lucas.
There can be concluded that the best method of the three is the long extraction method. 63,1% of the molds can be properly identified to the genus level with this method. Only 37% is correctly identified on the species level. These results aren’t reliable enough and on top of that there are still some misidentifications (15%). Besides that, the database is not comprehensive enough and the method is still too time-consuming.
Because of this the long extraction method won’t be implemented in the routine of the laboratory of AZ Sint-Lucas.
 
Samenvatting eindwerk 2013-2014: Validatie van 2 snelle methoden voor de identificatie van positieve hemoculturen met behulp van de Maldi Biotyper
Blood cultures are taken in patients with fever above 38 ° C or in patients with increasing temperature and signs of sepsis. Blood is normally sterile. If bacteria are present in the bloodstream, this indicates sepsis. In adults, at least 40 ml blood was drawn whereas in children only a pediatric vial (10 ml) is taken. When blood cultures forward in the lab, they are placed in the alert BacT/alert. This unit will detect the growth in culture flasks. In this way one can have positive and negative culture bottles apart. When the device detects a positive culture bottle, it is removed from the machine and microscopically examined by means of a direct examination and a Gram stain. Depending on the result of the Gram stain, other culture media and Combo panels for identification and antimicrobial susceptibility testing (AST) are deployed by Microscan. However, this method, time-consuming. In this method is the identification of the bacteria is present only after 16-18 hours. In order to shorten the time to identification, two methods are validate in this thesis. In both methods, the bacteria are identified by Maldi Biotyper. Only the steps for identifying varies in both methods. In the first method, the Sepsityperkit is used to extract the blood from the bacterial cells. In the second method, a drop of blood onto a Columbia agar with 5% sheep blood (COS). These COS blood plate is incubated in the CO2 incubator at 37 ° C for 4 to 8 hours depending on the type of bacteria. Validation is necessary to determine whether these methods exhibit a correct identification and thus can be used in the routine. From the results it appears that the plate method, yields better results than the method in which the kit is used. Also, the kit method is more expensive than the plate method. The kit costs 206 euros. Furthermore, the plate method is also less time-consuming method. It can thus be concluded that the plate method is a better alternative for the rapid identification of positive blood cultures.
 
Samenvatting eindwerk 2012-2013:Screenen op parasieten in feces door antigen-detectietesten
Het microscopisch onderzoek naar parasieten in feces is zeer arbeidsintensief, daarom kan er gezocht worden naar alternatieven die deze werklast verminderen.
In dit eindwerk wordt nagegaan of een sneltest goede resultaten levert om als screeningstest te gebruiken voor het opsporen van parasieten.
Eerst wordt er informatie gegeven over de te onderzoeken parasieten om zo duidelijk te maken waarom het zo belangrijk is voor de patiënt om een behandeling tijdig te starten.
Dan volgen de principes van de vier verschillende sneltesten (“Stick Crypto-Giardia”, “Rapid Strip Crypto-Giardia”, “RIDA® QUICK” en “Crypto-Giardia Duo Strip”), alsook hoe men deze moet uitvoeren.
Vervolgens worden de resultaten van de studie en beoordeling meegedeeld die helpen beslissen welke sneltest het beste scoort.
Uit de resultaten kan men concluderen dat de 4 sneltesten (“Stick Crypto-Giardia”, “Rapid Strip Crypto-Giardia”, “RIDA® QUICK” en “Crypto-Giardia Duo Strip”) zeer sensitief zijn. Bijgevolg kan men deze perfect gebruiken als screeningstest voor het opsporen van twee van de meest voorkomende parasieten; namelijk Giardia lamblia en Cryptosporidium parvum.

Address

St Lucaslaan 29
8310 Brugge
Belgium

Contacts

Traineeship supervisor
Johan Robbrecht
050/365346
johan.robbrecht@stlucas.be
Traineeship supervisor
Wouter Vandewal
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