To develop new diagnostic tests based on new DNA/RNA biomarkers, the DIAG4ZOO Team developed a molecular biosensor (mini-array) technology for biomarker monitoring and pathogen detection .
Based on PCR systems, the DIAG4ZOO Team developed a new technology that allows one-step multiple detections. When using this biosensor technology, the hybridization time is reduced to 15 min, whereas other methods require a longer hybridization time. Hybridization of the PCR product on a nylon membrane and revelation of the hybrids by an antibody considerably increase pathogen detection possibilities.
This biosensor method is based on a detection process combining target nucleic acid amplification by PCR and a specific hybridization evidenced by a colorimetric reaction on a nylon membrane. The hybrids are detected by using enzyme-linked antibodies producing a dark-blue precipitate with a substrate.
This method provides numerous advantages compared to other molecular diagnostic tools such as hybridization or amplification by PCR. These advantages include the possibility of a multiple detection, a higher sensitivity, a high specificity, no questionable results, etc. It is easy to use and results can be interpreted visually. Due to its reliability and high sensitivity, it can advantageously replace the common and tedious methods employed for evidencing amplification results obtained by electrophoresis. Another advantage is that it enables to avoid using Ethidium bromide which is a mutagenic reagent.
Multiple detections are one of the major advantages of this biosensor. In the case of multiple detections, multiple sets of specific primers are used and different amplicons issued from the PCR hybridize with the homologous probes plotted in pre-defined positions on the nylon membrane. The interpretation of the results observed on the membrane is easy and can be done by visual inspection. An internal control is added to each reaction. It is a PCR and hybridization positive control DNA fragment which is used to prevent false negatives due to experimental errors.
This technology substantially increases the power detection limit of pathogens and is a highly sensitive method contributing to virus prophylaxis as it is:
10,000 times more sensitive than the Dot Blot method.
100 times more sensitive than standard detection by electrophoresis of PCR amplified products.
It is the ideal solution for detecting low level or early infections in aim of limiting the risks of horizontal and vertical transmissions.
The test is rapid - one to 48 reactions can be performed at the same time in only 4 - 5 hours.
The membranes provided with the kit are ready to use. The interpretation of the results is easy - the presence or absence of blue dots can be detected visually. Little equipment is required - except for the thermocycler, only basic equipment such as a water-bath, a small bench centrifuge and a shaker are necessary. Therefore, this method can easily be used in farms or hatcheries already using PCR as a diagnostic tool.
If you require more information, please contact the DIAG4ZOO Team (Tel: +33 (0)467 419 748, Email: firstname.lastname@example.org).