Nationwide quality assurance of high-throughput diagnostic molecular testing during the SARS-CoV-2 pandemic: role of the Belgian National Reference Centre.

Since the onset of the coronavirus disease (COVID-19) pandemic in Belgium, UZ/KU Leuven has played a crucial role as the National Reference Centre (NRC) for respiratory pathogens, to be the first Belgian laboratory to develop and implement laboratory developed diagnostic assays for SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) and later to assess the quality of commercial kits. To meet the growing demand for decentralised testing, both clinical laboratories and government-supported high-throughput platforms were gradually deployed across Belgium. Consequently, the role of the NRC transitioned from a specialised testing laboratory to strengthening capacity and coordinating quality assurance. Here, we outline the measures taken by the NRC, the national public health institute Sciensano and the executing clinical laboratories to ensure effective quality management of molecular testing throughout the initial two years of the pandemic (March 2020 to March 2022).

Cross-evaluation of five slidemakers and three automated image analysis systems: The pitfalls of automation?

INTRODUCTION: Automated slidemakers and stainers and digital microscopes are coupled with haematology analysers to achieve better efficiency and cost-effectiveness. This study evaluates the integrated performance of slidemakers and digital microscopes commonly available on the market. METHODS: We compared the percentage of neutrophils for five slidemakers (two Siemens Advia Autoslides, a SysmexSP-10 and SP-50 and an Abbot Alinity hs) and a Horiba Hemaprep to the corresponding haematology analyser data (Siemens Advia 2120i, Sysmex XN and Abbot Alinity hq). Differential leucocyte counting (DLC) was performed on three different CellaVision digital microscopes (DM96, DM1200 and DI-60) and manually. The quality of the smears was assessed using a CellaVision SmearChecker. RESULTS: We observed a significant positive absolute bias (P < .05) for the percentage of neutrophils with the Autoslide and Alinity hs smears on the digital microscopes, but not when DLC was performed manually. The SP-10 and SP-50 showed no bias regardless of the DLC method. No bias was observed for the Hemaprep smears. All the smears had an acceptable monolayer quality, stain intensity and colour. All smears, except those from Sp-10, were of an acceptable length. CONCLUSION: Users should be aware of a potential lack of accuracy that can be encountered when using some slidemakers and digital microscopes. All laboratories should validate or verify the differential counts from slidemakers and digital microscope with automated cell differential counters. Manual count validation should only be considered if a significant proportion of clinically relevant abnormal cells are present. Otherwise, haematology analyser results should be favoured.

Up-regulation of the Yersinia enterocolitica yop regulon by deletion of the flagellum master operon flhDC.

The Yop virulon enables extracellularly located Yersinia, in close contact with a eukaryotic target cell, to inject bacterial toxic proteins directly into the cytosol of this cell. Several Ysc proteins, forming the Yop secretion apparatus, display homology with proteins of the flagellar basal body. To determine whether this relationship could extend to the regulatory pathways, we analyzed the influence of flhDC, the master regulatory operon of the flagellum, on the yop regulon. In an flhDC mutant, the yop regulon was up-regulated. The transcription of virF and the steady-state level of the transcriptional activator VirF were enhanced. yop transcription was increased at 37 degrees C and could also be detected at a low temperature. Yop secretion was increased at 37 degrees C and occurred even at a low temperature. The Ysc secretion machinery was thus functional at room temperature in the absence of flagella, implying that in wild-type bacteria, FlhD and/or FlhC, or the product of a gene downstream of flhDC, represses the yop regulon. In agreement with this notion, increased expression of flhDC in wild-type bacteria resulted in the oversecretion of flagellins at room temperature and in decreased Yop secretion at 37 degrees C.