Applications for real time pcr market survey


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Applications for real time pcr market survey

  1. 1. PARIOFORMA Summary Report – September 2010PARIOFORMA LTD55 Princes GateExhibition RoadLondon SW7 2PNUnited KingdomTel: +44 (0) 20 7225
  2. 2. PARIOFORMA Since its introduction little more than 10 years ago, real-time PCR has become the main technical platform for nucleic acid detection in research and development. The scope of real-time quantitative (qPCR) assays for gene-expression analysis in research applications and for sequence-specific nucleic acid detection for diagnostic purposes continues to grow. Growth drivers include: ◦ Speed ◦ Increasingly robust assays ◦ Automation ◦ Higher-throughput ◦ Multiplexed analytical strategies ◦ Increasingly affordable machines Technology trends have focused on: ◦ Increased throughput ◦ Faster analysis time Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 2
  3. 3. PARIOFORMA ◦ Quantitative resolution ◦ Assay kits – specific assays for known targets – (now even with the probe and primer sequences to facilitate compliance with evolving MIQE guidelines). ◦ Ready to use reagents ◦ Automation/ pre-analytical sample processing ◦ User friendly interfaces:  Software driven inputs/ controls – “menu” driven devices  Data output - analysis software providing amplification curves, color-coded heat maps, cycle threshold (Ct) data, standard curves, etc. for each run. ◦ Point of test/ point of care testing Real-time PCR is becoming a mature technique. Instruments are very robust and reliable, and optimised assays can be ordered from multiple vendors. Although the technique will be further refined and more niche products will be developed, main future improvements are not expected within the real-time PCR technology itself. Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 3
  4. 4. PARIOFORMA Real-time monitoring of PCR has simplified and accelerated PCR laboratory procedures and has increased information obtained from specimens including routine quantification and differentiation of amplification products. Fast, quick and reliable results are expected of molecular diagnostics - early diagnosis allows for appropriate and timely clinical intervention. Rapid or "almost point-of-care" molecular tests take about a few hours versus days for a culture- based test. Clinical diagnostic applications and uses of real-time PCR are growing exponentially, real-time PCR is rapidly replacing traditional PCR, and new diagnostic uses continue to emerge. Real-time PCR was first introduced for infectious disease testing and kits are offered today from a variety of suppliers: - e.g. Roche, Abbott, Qiagen, Biotools, Roboscreen and LightUp Technologies. All assays are probe-based to minimize the rate of false positives. Assays for targets with high mutation rates, such as HIV and HCV are optimized for subtypes and clades prevalent in developed countries, and may have poor sensitivity for the variants common in the parts of the world were these tests are most needed. Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 4
  5. 5. PARIOFORMA In vitro diagnostic tests may be segmented: ◦ Commercial in vitro diagnostic products ◦ Diagnostic assays developed by laboratories develop themselves - so called "home brew" assays . There is a high cost coupled with a considerable time lag for commercial kits to gain regulatory approval and be available on the market. Consequently, for some infections there may be no commercial interest in developing FDA-approved assays. Much initial diagnostic work is done with home brew assays - 2003 definition/ guidance from the FDA: ◦ Assays incorporate “general purpose reagents [and] general purpose instruments [used] by a laboratory to set up an in-house (“home brew”) test or laboratory testing service. ◦ While specimens can [and do] travel to the laboratory setting up this service, the test itself is not marketed outside of the single laboratory setting up this service. ◦ It is the responsibility of the laboratory to develop a recipe for the [home brew] test at hand and to take responsibility for establishing and maintaining performance. ◦ Although not part of the [ASR/home brew] rule, FDA has indicated that FDA is not requiring the in-house test to go through premarket FDA review. See FDA, CDRH, Analyte Specific Reagents. Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 5
  6. 6. PARIOFORMA In Europe, assay systems/ kits need to be CE marked under the European In Vitro Diagnostics Directive – but in-house (home brew) assays fall outside of the scope of the Directive. However, if use of the in-house assay becomes part of a commercial transaction then the Directive ”should” apply. Market feedback demonstrated that in some countries home brew assays are reimbursed for clinical use.Multiplexing There has been a trend to increase the multiplexing capabilities of qRT-PCR analysis. Instrument manufacturers were aware that limitations on the number of channels capable of measuring different mRNA targets had to be addressed in order to allow for qRT-PCR to analyze the full spectrum of interacting mRNA signals resulting from a cellular response. New instruments have been brought to market in the past 5 years installed with many more filter channels capable of complex multiplexing. Market feedback suggests that multiplexing is not without its difficulties in terms of experimental design, setup and in particular reproducibility. Nevertheless, end users increasingly demand machines with multiplex capabilities even though more often than not, they are only using duplex functionality. Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 6
  7. 7. PARIOFORMA In the R&D industry (includes academic but primarily the private sector of pharmaceutical/ biotechnology companies), qRT-PCR has become a valuable tool utilized to understand the response to different treatment options by monitoring the various signalling pathways present in cells. An early inhibitor to the utility of qRT-PCR in the R&D sector was that end-users were being hindered by the time and effort necessary to design and optimize their experiments. However, this has been overcome in the main by the development of: ◦ Sophisticated software solutions aimed at experimental design and setup (e.g. primer design) ◦ Higher quality reagents ◦ “Off-the-shelf” kits Today the major concerns surround: ◦ Standardisation of sample management ◦ Data management This is a very high throughput and high-volume segment - in addition, this is a crowded segment that attracts the largest of market players each chasing the R&D “cash cow.” Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 7
  8. 8. PARIOFORMA The global market for food safety testing can be broken down by contaminant type and includes: ◦ Pathogens ◦ Genetically modified organisms (GMOs) ◦ Toxins ◦ Residues ◦ Other contaminants / ingredient verification Pathogen testing has the largest share of the market followed by GMO testing. Drivers for food testing include: ◦ Recurring food-related public health issues ◦ Consumer expectations ◦ Mounting regulatory food safety requirements ◦ Industry competitive advantage ◦ Cost of product recall ◦ Protection of brand value ◦ Avoid litigation Many companies do not recover financially from a food-borne illness outbreak associated with their product. Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 8
  9. 9. PARIOFORMA Historically, pathogen reduction efforts focused on the pre-release screening of finished product – however, this approach does little to prevent processing failure or improve production efficiency. Today the focus is on the entire processing continuum. This shift in focus has not only resulted in a measurable reduction in the incidence of food-borne pathogens but has also significantly reduced the need for extensive finished product screening and costly release holds. Drivers for adoption of real time PCR in food testing: ◦ The main driver for PCR systems over other methods is in time saving, both in the total time from sampling to result and in the technician time needed to set up and run the assay. ◦ High degree of automation built into PCR systems means that staff time and training is effectively minimised. ◦ High specificity generally means fewer repeat tests are required. ◦ Availability of commercial kits with AFNOR and/or AOAC validation. ◦ Multiplex capabilities allowing for several pathogens to be assayed in a single run. ◦ Real-time PCR assays can also be made quantitative without adding additional stages or operations. ◦ Cost benefits in rapid test results. Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 9
  10. 10. PARIOFORMA Drivers for GMO testing: ◦ Controversies surrounding human and environmental safety ◦ Labeling and traceability ◦ Consumer choice ◦ Intellectual property rights ◦ Food security Efficient detection of genetically modified organisms is becoming increasingly important to comply with more restrictions and increased awareness in food safety. The presence of transgenic crops in the food and feed market has given rise to different regulations in different countries – for example: ◦ US: food labelled “GM free” if <5% GM ◦ EU: food labelled “GM” If > 1% GM ◦ Japan: food labelled “GM” If > 5% GM PCR is the favoured method for detecting and quantifying GMOs. However, criticism of the technique remains in that it is costly, highly technical, time consuming, and often wavers in accuracy and precision. Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 10
  11. 11. PARIOFORMAParioforma Ltd. Is an independent business consultancy headquartered in London, UK.Key benefits of working with Parioforma: Bespoke studies – tailored to client needs and timelines. LONDON OFFICE ASSOCIATE OFFICES Affordable research prices – makes your budgets go further. Intrix Corp, Darien Charles Rowlands USA High quality research – experienced science-based Connecticut team, all with post-graduate science and/ or MBA Senior Partner qualifications and with 10-25+ years experience in Life Eastern Science and High Technology industries. 55 Prince’s Gate IMRP, Moscow Europe Unique problem solving approach – the team is Exhibition Road highly specialised, but diverse in experience and London SW7 2PN Intrix Corp, São background, enabling analysis of problems from Brazil United Kingdom Paulo different perspectives. Tel: +44 (0) 7803 907577 Strategic Pointers – helps clients better understand Japan TRN, Tokyo the research and implications for their business. Practical implementable solutions – not just “over-clever analysis.” This is a sample report only. Please dontOur knowledge is extensive and experience hesitate to contact us for an informal discussionbroad – please visit our website for a full on your market research needs – either bylisting of all our services. telephone on +44 (0) 207 225 3538 or by email at: Parioforma Ltd – 55 Princes Gate – London SW7 2PN – United Kingdom 11