Final Report Summary - PORT CHECK (Development of generic 'on site' molecular diagnostics for EU quarantine pests and pathogens)
The project was a combined RTD and demonstration activity between a number of RTD laboratories, SME laboratories and end user laboratories (NPPO) with the aim of developing and testing a portable real-time Polymerase chain reaction (PCR) testing platform for some key EU quarantine organisms. PORT CHECK aimed to deliver the tools and procedures to allow EU Member State plant health competent laboratories and inspection services to perform molecular diagnostic assays 'on-site' and at points of entry.
The project developed and evaluated real-time PCR (TaqMan and CyCleave) assays for a number of key harmful organisms, including the sudden oak death pathogen (Phytophthora ramorum) and the pinewood nematode (Bursaphelenchus xylophilus); and transferred these assays to field portable real-time PCR platforms.
The PORT CHECK project consortium performed R&D to develop every part of the process involved in on-site detection. From sampling and sample homogenisation, through simplified DNA extraction, set up of real-time PCR detection reactions using field-stabilised reagents to actual testing and interpretation of the results. The protocols developed were validated using infected material and were compiled into a comprehensive publicly available booklet for the detection of 12 different quarantine pathogens and pests.
By working with SME companies prototype equipment and kits were made available to the consortium (and post project to the wider community) to enable homogenisation of samples, DNA extraction, field stable reagents and portable real-time PCR. PORT CHECK then put the technology in the hands of thirty-two NPPO laboratories and inspection services for evaluation. The results from the method evaluation study showed that non-specialist users could generate results equivalent to those achieved in the laboratory in less than two hours, and that this could be done remotely from the laboratory.
The project developed and evaluated real-time PCR (TaqMan and CyCleave) assays for a number of key harmful organisms, including the sudden oak death pathogen (Phytophthora ramorum) and the pinewood nematode (Bursaphelenchus xylophilus); and transferred these assays to field portable real-time PCR platforms.
The PORT CHECK project consortium performed R&D to develop every part of the process involved in on-site detection. From sampling and sample homogenisation, through simplified DNA extraction, set up of real-time PCR detection reactions using field-stabilised reagents to actual testing and interpretation of the results. The protocols developed were validated using infected material and were compiled into a comprehensive publicly available booklet for the detection of 12 different quarantine pathogens and pests.
By working with SME companies prototype equipment and kits were made available to the consortium (and post project to the wider community) to enable homogenisation of samples, DNA extraction, field stable reagents and portable real-time PCR. PORT CHECK then put the technology in the hands of thirty-two NPPO laboratories and inspection services for evaluation. The results from the method evaluation study showed that non-specialist users could generate results equivalent to those achieved in the laboratory in less than two hours, and that this could be done remotely from the laboratory.
