Legionella pneumophila SG1 Kit for the CellStream Ultrafast Detection Including Viability Assessment Speed Separation, concentration and purification of Legionella pneumophila in 1-2 hours Specificity High specificity and sensitivity Superior monoclonal antibody pools Detection of viable but non-culturable Legionella pneumophila Easy handling Flexible integration into any workflow Compatible with flow cytometry, PCR, cultivation method, microscopy, etc.
rqmicro developed this highly specific kit to simplify and accelerate the detection of Legionella pneumophila SG1 in water samples. Together with the CellStream it opens a new chapter in the field of pathogen detection and in the monitoring of water quality. Immunomagnetic Separation and Microfluidics The sample passes through a narrow channel in the microfluidic cartridge. During this process, a magnet is lowered down and thereby immobilizes the bacterial cells, which are bound to magnetic particles via specific antibodies. At the same time over 95% of the competing flora, originally present in the sample, is directed to the residual fraction. In the final step of the separation, the magnet is raised and the target cells, together with clean buffer, are directed into the positive fraction. Flexible Integration into Your Workflow Concentration Labelling Automatic Separation on Disposable Cartridge Analysis Quantitative results within 1-2 hours following a straightforward workflow from filtration to quantification using standard flow cytometry (FCM). rqmicro AG Page 2
Rapid Sample Preparation for Flow Cytometric Analysis Red Fluorescence Red Fluorescence rqmicro s kits are suitable for all standard flow cytometers. Using FCM in combination with the CellStream, water laboratories gain new insights into water systems due to specific live and dead staining of pathogens and the possibility of monitoring processes in real time, e.g. disinfection. Green Fluorescence Green Fluorescence Example of a flow cytometric quantification of Legionella pneumophila SG1 cells after the immunomagnetic separation with the CellStream before (left) and after (right) a successful disinfection. The sample is stained with staining dye (green) and viability dye (red). In the dot plot on the left, Legionella pneumophila cells are green. The cells are viable and possess intact membranes. Therefore, the viabiliity dye is not able to enter the cell and the cells don t get stained in red. On the dot plot on the right, the Legionella pneumophila cells are red and green. The disinfection process destroyed the integrity of the membrane and the cells are no longer viable. Pick the Analytical Method of Your Choice In addition to flow cytometry, the CellStream supports other analytical methods (e.g. cultivation method, PCR, fluorescence microscopy). Contact our experts for more information. rqmicro AG Page 3
Performance Sample purity: 95-99.9 % Limit of detection: 20-50 cells / liter* Background noise: 0-25 events / liter* Separation time: 35 minutes *Dependent on flow cytometry instrumentation used Detected L.pneumophila cells/liter Performance tests show a recovery of > 80 % and a broad working range. Triplicates per data point. The lower detection limit is at around 20-50 cells per liter. Spiked L.pneumophila cells/liter Equivalence to ISO Method Cultivation on BCYE; detected L.pneumophila-colonies/mL The rqmicro testing procedure in combination with flow cytometry achieves > 98 % correlation with the current gold standard (ISO method). The sensitivity of the rqmicro method is 5-10 fold higher. Eight measurements per data point. Equivalent or more accurate results achieved in 1-2 hours instead of in 10 days. rqmicro testing procedure; detected L.pneumophila cells/ml rqmicro AG Page 4
Specificity Cross reactivity experiments with the rqmicro Legionella pneumophila SG1 Kit using various SG1 strains portray high specificity and a complete coverage of SG1. Additional cross reactivity experiments are ongoing. For up-to-date information, please visit www.rqmicro.com. Organism L.p. SG1 Antibody binding Legionella pneumophila Philadelphia Legionella pneumophila OLDA Legionella pneumophila Knoxville Legionella pneumophila Bellingham Legionella pneumophila Benidorm Legionella pneumophila Oxford Legionella pneumophila Allentown/France Legionella pneumophila Camperdown Legionella pneumophila Heysham DSM 7513 DSM 25200 DSM 25070 DSM 25214 DSM 25199 DSM 25213 ATCC 43106 ATCC 43113 ATCC 43107 No Cross Reactivity Extensive tests confirmed the specificity of the Legionella pneumophila SG1 Kit and evidenced no cross reactivity with the following, relevant organisms from the water and food industry. An up-to-date list is available on www.rqmicro.com. Legionella pneumophila SG 2-14 Legionella anisa Legionella birminghamensis Legionella bozemanii Legionella cincinnatiensis Legionella dumoffii Legionella jamestowniensis Legionella jordanis Legionella londoniensis Legionella longbeachae Legionella micdadei Legionella oakridgiensis Legionella parisiensis Legionella parisiensis Legionella rubilucens Legionella taurinensis Pseudomonas aeruginosa Pseudomonas fluorescens Staphylococcus aureus Staphylococcus epidermidis Campylobacter jejuni Escherichia coli Salmonella typhimurium Citrobacteri gillenii Listeria monocytogenes Listeria welshimeri Listeria fleishmanii rqmicro AG Page 5
90% Downtime Reduction Disinfection procedures guided by rapid tests: are targeted save costs and energy result in less downtime for affected infrastructures Rapid testing method provides immediate process feedback Enables operators to take corrective measures and/or stop process once complete Accurate Detection of VBNC Bacteria Real time monitoring of a disinfection process at 70 C Viable but non-culturable (VBNC) bacteria refers to bacteria that are in a state of very low metabolic activity. They do not divide, but are alive and have the ability to become culturable once resuscitated. rqmicro s testing procedure reveals that after a disinfection process viable Legionella cells are potentially still present after 60 minutes. FCM Cultivation Method The cultivation method does not detect VBNC bacteria. The presence of VBNC bacteria does therefore question the accuracy of the current gold standard (ISO method). rqmicro AG Page 6 VBNC
Application L.p. SG1 Kit L.p. SG1 Kit PLUS Cooling Tower Water Surface Water Drinking Water Who Can Benefit from rqmicro s Rapid Legionella Test Cooling tower operators Environmental service laboratories Healthcare Institutions (hospitals, nursing homes, etc.) Industrial plants Public facilities (schools, universities, etc.) Water supply companies Public swimming baths, wellness and spa sector, etc. Kit Components (96 tests) Cartridges: Disposable microfluidic cartridges Magnetic particles: Magnetic particles, conjugated to a monoclonal antibodies Staining dye: Labelling antibody; Ex/Em 493/518 nm Viability dye: Propidium iodide as an indicator of membrane integrity; Ex/Em 535/617 nm Positive control: Lyophilised, chemically fixed, freeze-dried Legionella pneumophila SG1-cells (Philadelphia) Filters: Polycarbonate filters; pore size 0.22 µm; Ø 47 mm Cell strainers: CellTrics filters ; pore size 30 µm (SG1 Kit PLUS) Buffer 1: Washing buffer Buffer 2: Separation buffer Current Test Kits in Development Our R&D team is constantly working on expanding our offer. To check our current kits in development and our news please check out our website: www.rqmicro.com rqmicro AG Page 7
Contact us: rqmicro AG Brandstrasse 24 8952 Zürich - Schlieren Switzerland Tel: 41 44 512 51 51 Sales support: sales@rqmicro.com General information: info@rqmicro.com www.rqmicro.com Copyright rqmicro AG 2017 Test Kit SG1 Leaflet 201710