Matrix-assisted laser desorption/ionisation time of-flight mass spectrometry (MALDI-TOF MS) is a fast and reliable method for the identification of bacteria from agar media. Direct identification from positive blood cultures should decrease the time to obtaining the result. In this study, three different processing methods for the rapid direct identification of bacteria from positive blood culture bottles were compared. In total, 101 positive aerobe BacT/ALERT bottles were included in this study. Aliquots from all bottles were used for three bacterial processing methods, i.e. the commercially available Bruker's MALDI Sepsityper kit, the commercially available Molzym's MolYsis Basic5 kit and a centrifugation/washing method. In addition, the best method was used to evaluate the possibility of MALDI application after a reduced incubation time of 7 h of Staphylococcus aureus- and Escherichia coli-spiked (1,000, 100 and 10 colony-forming units [CFU]) aerobe BacT/ALERT blood cultures. Sixty-six (65%), 51 (50.5%) and 79 (78%) bottles were identified correctly at the species level when the centrifugation/washing method, MolYsis Basic 5 and Sepsityper were used, respectively. Incorrect identification was obtained in 35 (35%), 50 (49.5%) and 22 (22%) bottles, respectively. Gram-positive cocci were correctly identified in 33/52 (64%) of the cases. However, Gram-negative rods showed a correct identification in 45/47 (96%) of all bottles when the Sepsityper kit was used. Seven hours of pre-incubation of S. aureus- and E. coli-spiked aerobe BacT/ALERT blood cultures never resulted in reliable identification with MALDI-TOF MS. Sepsityper is superior for the direct identification of microorganisms from aerobe BacT/ALERT bottles. Gram-negative pathogens show better results compared to Gram-positive bacteria. Reduced incubation followed by MALDI-TOF MS did not result in faster reliable identification.
Publicatie bij de rede, uitgesproken bij de aanvaarding van het ambt als lector Green Biotechnology aan Hogeschool Inholland te Amsterdam op 20 mei2015 door dr. C.M. Kreike
Every healthcare professional (HCP) in the Netherlands is expected to provide palliative care based on their initial education. This requires national consensus and clarity on the quality and goals of palliative care education and accessible education opportunities nationwide. These requirements were not met in the Netherlands, posing a major obstacle to improving the organization and delivery of palliative care. Therefore, a program, Optimizing Education and Training in Palliative Care (O2PZ), was established to improve palliative care education on a national level.
MULTIFILE
New innovative methods to determine the DNA sequences of different bacterial species are rising. In the field of microbiology, these methods are very important since it is now possible to determine all the genetic characteristics of the bacterium in one step! This enables to define e.g. the species family, drug resistance or relatedness to other bacteria in outbreak evaluations which is necessary to efficiently treat the bacteria or target potential outbreaks. For many years, PCR-based methods have been the technique of choice to determine DNA sequences (including next-generation sequencing techniques). Recently, a new technique has been introduced to the market that is based on single molecule real-time sequencing (SMRT) with the possibility to determine the DNA sequence of a bacterium. This SMRT MinION sequencing technique is housed on an USB stick and is known for its user-friendliness and huge data output. However, before such a new technique can be implemented and presented in laboratories and used for educational purposes, methods should be harmonized and evaluated to proof its applicability. Harmonisation of the methodology regarding new laboratory techniques is very important to be able to compare results generated by different laboratories. A single consistent protocol, applied in each lab, is essential to obtain the best results in interlaboratory comparisons. During this KIEM-hbo project, we – i.e. Avans UAS, Maastricht University Medical Center and the company IS-diagnostics – will determine the DNA sequence of bacterial species and mixes thereof with a harmonized protocol for an interlaboratory comparison. We will compare this technique to the IS-PRO, an existing technology. Finally a workshop will be organized for medical technicians and other SMRT sequencing users to evaluate the protocols. This will, generate an up-to-date and harmonized sequencing protocol which can be expanded to future research and diagnostics in the different areas.
