Parioforma Ltd. is a London-based business consultancy that provides market research services. This report summarizes a study on quantitative real-time PCR (qRT-PCR) technology and market trends. Key findings include that qRT-PCR is a mature technique seeing growth in automation, high-throughput applications, and standardized assays. While instruments are reliable, future improvements may focus more on sample preparation and data analysis to support the large amounts of data generated.

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Summary Report – September 2010
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 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 qRT-PCR technology itself.
 Growth drivers include:
◦ Increasingly robust assays
◦ Automation
◦ Increasing demand for high-throughput
◦ Multiplexed analytical strategies
◦ Increasingly affordable machines
 Technology trends have focused on:
◦ Increased throughput
◦ Faster analysis time
◦ Quantitative resolution
◦ Ready to use reagents and assays – including probe and primer sequences to facilitate compliance with
evolving MIQE guidelines
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◦ Automation:
 Sample processing
 Liquid handling & filling
− User friendly instrument operation:
 Software driven controls – “menu” driven devices
 Sophisticated data analysis software – integration with existing LIMs
 Future challenges will be to improve the down and upstream steps of real time PCR:
◦ Downstream:
 Reagents: quality, specificity and consistency
 Sample management and preparation
◦ Upstream:
 The challenge is to handle, annotate and analyse the very large amounts of data that will be generated.
 The implication is increasing sophistication of dedicated software for real-time PCR quality control, data
management and analysis.
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 For existing qRT-PCR users factors limiting the maximum use of the technology include:
– The cost of instrumentation and consumables.
– Machine access - more than 70% of scientists often share instrumentation with 10-12 other colleagues.
 Factors inhibiting the uptake of qRT-PCR by new users identified in interviews include:
– The perceived difficulty and complexity of running qRT-PCR assays
– Lack of experience in the technique
– Perceived cost of instrumentation and reagents – although existing methods take longer, they are cheaper to
run
– Barriers to change existing working practices
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 Early adopters include: Real time PCR is a mature technique but not
Biotech & Pharma; a mature market
Academia and Government
research. These are well
established in their use of
Saturation level
Saturation level
Saturation level
Saturation level
Saturation level
Saturation level
GMO
qRT-PCR with a large Forensics
Biodefense
installed machine base.
Smaller food labs
Large food Labs
 Late adopters include: Veterinary
Clinical Labs
forensics, veterinary Academia Diagnostics
diagnostics. These are just
Big Large
beginning to acquire Roche Pharma CROs
machines and develop new Biotech
ABI
methods.
 Others are somewhere in
between – e.g. hospital &
clinics; food & GMO testing;
environment testing. These
are expanding existing use The diffusion of innovation technology With successive groups of consumers adopting
of qRT-PCR into new areas. the new technology (shown in blue), its market share (yellow) will eventually reach the
saturation level.
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 Primary application areas for qRT-PCR analyses are basic research and diagnostics (detecting the
presence of target genetic material) – others are derivatives thereof:
Basic Research Diagnostics
In research settings, qRT-PCR is mainly used to provide Molecular diagnostics on the other hand, involves the
quantitative measurements of gene transcription – or measurement of DNA, RNA, proteins or metabolites to
gene expression which is the process by which detect genotypes, mutations or biochemical changes.
information from a gene is used to synthesize a Diagnostic qRT-PCR is applied to rapidly detect nucleic
functional gene product (e.g. a protein). acids that are diagnostic of, for example, infectious
The technology may be used in determining how the diseases, cancer and genetic abnormalities –in humans,
genetic expression of a particular gene changes over plants and animals, etc.
time, such as in the response of tissue and cell cultures The introduction of qRT-PCR assays to the clinical
to an administration of a pharmacological agent, microbiology laboratory has significantly improved the
progression of cell differentiation, or in response to diagnosis of infectious diseases, and is deployed as a
changes in environmental conditions. tool to detect newly emerging diseases, such as flu, in
Gene expression profiling can help researchers diagnostic tests.
understand disease pathways which in turn helps them
to identify means to interfere with those disease
pathways (e.g. pharmacological) to develop treatments.
Another important research application is genotyping.
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THROUGHPUT
 The most widely used assay format is 96-well – “96-well” although not set as standard has become
the industry standard across all applications.
 Below 96-well is considered “low throughput” – users in this sector are looking for flexibility in
sample numbers per run – e.g. as provided by the Roche LightCycler® 2.0 Instrument (1 – 32
samples per run)
 The second most widely used assay format was found to be 384-well pcr plate.
ASSAY VOLUME
 The majority of respondents were using a total assay volume range 10-25μl.
 A preference for 5μl of sample in a overall sample volume of 25μl was identified.
 The second most widely used assay volume range was found to be 25-50μl.
 End users in diagnostics (food, clinical, veterinary) all indicated a need for higher sensitivity and
higher overall reaction volumes – in these applications precision and accuracy are paramount.
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AUTOMATION
 Laboratories employ automation as a solution to achieve higher throughput and increased walk-
away time – an automated solution must deliver improved productivity and workflow.
 Key needs for automated solutions:
◦ Cost-effective
◦ Fully-scalable
◦ Simplifies processes within the lab without compromising accuracy and precision
◦ Natural fit to lab work-flow (no added steps)
◦ Plug-and-play – software driven
◦ Bench-top
◦ Minimal manual reloading
 Interview findings suggest the end-users most well established in the use of automation are the
early adopters of qRT-PCR (Pharma & Biotech R&D; Academia; Government research) – these have
applied automation to many of their high throughput lines (>96 well).
 Adoption of automation outside of these segments is patchy and tends to follow how well
established the lab is in using qRT-PCR (more frequent in larger labs than smaller ones).
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REAGENTS
 The majority of respondents indicated a preference for pre-optimized qRT-PCR mastermixes (used
according to manufacturers instructions) as opposed to qRT-PCR bulk reagents bought separately
(optimized & validated in own lab).
 End-users want reagent systems that work first time without optimisation or validation.
 A large majority of consumables budget is spent on commercial kits – nevertheless, it must be
clarified that end users are budget conscious. Many standard test kits are now available at very
attractive prices while others (notably food and some diagnostic assays) command very high prices.
 End users tailor the use of commercial and in-house (home brew) assays to suit their budget and
productivity needs:
 Sometimes it is more cost effective to buy a test (even if expensive) rather than develop and validate an in-
house system (typical in large food labs);
 However, many large, busy clinical labs are now finding it more cost effective to develop in-house systems
for high volume, routine analysis.
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REAGENTS – immobilized systems
 The majority of Interviews revealed a high level of interest in lyophilized reagents:
Advantages Disadvantages
Convenient and time saving Expensive
Easy to use Potential for wastage
De-skills end user staff requirements Storage and shelf-life concerns
Proven to improve quality and reliability of the End users dislike instrument systems with “tied”
qRT-PCR reaction assays
 It was reported that there is increasing demand for lyophilized systems – but the market remains
embryonic with a limited number of suppliers which includes the large recognised names as well as
emerging niche suppliers.
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 Data quality expectation is the major buying selection criteria.
 Interview feedback indicated that end users buy “brands” – where the brand name conveys quality
and confidence in instrument performance.
 Other important selection criteria included:
◦ Data analysis capability features
◦ Assay design
◦ HRM capability
◦ Running costs – cost of consumables
◦ Past experience
◦ Availability of training
 Instrument cost was cited as being less important overall than the aforementioned criteria.
 The instrument market is crowded – success depends on offering a quantum leap in features and
benefits as perceived by the end-user:
◦ “It is very difficult for any newcomer to come to market with a “me-too” device and grab significant market
share. Market penetration occurs by word of mouth – one lab has one and another will want it!” – Director of
Research, Medical Genomics.
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