The ProWaChip project will develop a Bio-Chip for fast, on-site detection of bacterial contamination in drinkable and industrial water, whose detection principle is based on a high-specific hybridization reaction taking place within the microfluidic chip. The BioChip performs both the lysis and analysis of bacterial RNA in a single substrate. The BioChip will be developed to produce “On-line”, “in-situ” detection of two indicator bacteria (E. Colli and L. pneumophila), technique which constantly monitors the process water of industries, such the sugar production, and becomes functional in drink water systems in combination with AGIL-equipment.
Currently, the lack of suitable control methods for microbiological inspection represents an obstacle for an “in-situ” controllability of hygienic quality of process and drink water. On the other hand, the identification of sub-lethal or damaged bacteria by traditional microbiology methods is limited due to their restricted cultivation. In order to be independent of such limitations, a BioChip is developed to process minute volume of sample and reagents and to enable “On-line”, “in-situ” monitoring of running water. The biosensor will consist on a microfluidic system and exclusively detect alive bacteria by means of reaction on specific nucleic-acid. To be able to assess a concrete load of alive bacteria, the short-lived bacteria RNA molecule will be used as hybridization target. Due to its rapid degradation, the RNA can only be identified during a short time after the lysis is conducted over alive bacteria. The RNA releasing lysis consists on the disruption of the bacterial membrane by means of, e.g. enzymes or heat. The released RNA will be directed by fluid stream into the Analysis area of the BioChip and bound to nucleid-acid capture probes, which are arranged over the reaction chamber surface. Subsequently, the captured RNA will be hybridized with fluorescence labelled probes, and by this way their presence becomes identifiable by an optical sensor.
The BioChip is planned as a disposable product to fulfil hygiene and bio-safety regulations. This requirement motivates the BioChip fabrication using a low-cost, biocompatible and low auto-fluorescence material. The lysis and analysis functions will be integrated and carried on within the same device. Due to the requirements of microbiology protocols, the system should process up to 1 mL of sample water. The overall processing time is expected to be 1 hour as maximum.
The BioChip will be conceived as a low-cost, disposable device. Therefore it will be manufactured of a moldable, low auto-fluorescence polymer such as Cyclo-Olefin Copolymer (COC).
Up to 1 mL of sample water within 1 hour will be processed. Hence the volume of BioChip microfluidics and reagents must be planned.
For implementation of Lysis, an integrated micro-heater will warm up the sample water up to 95°C. However the same sample water volume must be available at the Analysis chamber with a temperature no greater than 55°C. As consequence, fluid cooling micro-channels and integrated temperature sensors for controlling the heater must be implemented.
BIAMOL (Bremerhavener Institut für angewandte Molekularbiologie) – Hochschule Bremerhaven
Junker Filter GmbH
APM Gehäusetechnik GmbH
Prof. Dr.-Ing. Walter Lang
IMSAS, NW1, Room O2120
Tel: +49 421 218 62602
Teh project was funded from June 2015 for three years by AiF Projekt GmbH as a cooperation project within the frame of the "Zentrales Innovationsprogramm Mittelstand (ZIM).