Tag Archives: Testing

Ravi Ramadhar, Food Safety Business Director for Life Sciences Solutions, Thermo Fisher Scientific
In the Food Lab

Molecular Diagnostics – Generation 3: 2005 to Present

By Ravi Ramadhar
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Ravi Ramadhar, Food Safety Business Director for Life Sciences Solutions, Thermo Fisher Scientific

In my previous blog, I covered the first two generations of Molecular Diagnostics: Generation one, was the advent of these tests prior to 1995, while the second generation saw the evolution of molecular diagnostics with the emergence of standardized food molecular and method workflow.

The advent of automated DNA sequencing and use of multiple fluorescent dyes by companies like Applied Biosystems and Roche led to the development of multiple fluorescent dyes and real- time quantitative PCR systems (qPCR). At first these qPCR systems were only used in the research environment, but quickly found their way to the food industry.

Applications such as quantitation of GMOs and multiple pathogen targets became common. Real-time PCR systems permitted users to visualize amplification as it happened and enabled simultaneous detection of multiple targets. With the use of newer chemistries and improved enzymes, shorter amplification cycles – sometimes as low as 40 minutes – could be achieved. The real-time systems offered faster time-to-result with additional target probes and thus higher target specificity. As with most molecular methods, the workflow was sensitive to food matrix inhibition and required alternative sample preparation methods to meet the wide variety of food matrixes.

Within this generation of solutions, alternatives were introduced, that promised faster, easier or more sensitive results. These included alternative to either the detection method or enzymes utilized Iisothermal amplification, for example without need for multiplexing capability of qPCR or internal controls, as well as targeting alternative nucleic acid such as RNA were introduced to the food market. These incremental improvements did not lead to any significant new paradigms or improvements to the food testing workflow. Their emergence instead led to an explosion of additional and alternative molecular platforms for food, without any real innovation. Within this, solutions introduced to the food industry eventually brought us to where we are today.

Directly taking systems from the clinical diagnostics workflow and introducing these platforms and systems as food solutions. While these systems automate the entire workflow or automate the PCR setup it remains to be seen if with their higher complexity and high maintenance these systems can survive the food industry. The basic molecular workflow for food has remained intact since its introduction in the late 1990s with innovation more or less stagnant. What’s needed is for someone to truly develop a platform from the ground up with the food laboratory in mind.

Today’s landscape and what’s next

Today, there are some early signals of where innovations and changes for food labs will emerge. A recent poster by Nestle, for example, highlighted the uses of next-generation sequencing (NGS) and DNA sequencing to develop a DNA method to allow the identification of coffee varieties through the value chain, from the field to the finished product. The method is applied on routine basis to guarantee the purity and authenticity of raw material used by Nespresso.

Applications of NGS in outbreak response and trace back investigations are being used in parallel with existing technologies. Finally, availability of new sequencing data enables better assay design and development of adjacent technologies.

NGS was preceded by emulsion amplification and sequencing by synthesis. These developments led to the development and introduction of digital PCR. Within a digital PCR reaction, millions of simultaneous reactions from one sample occur. The advantages of dPCR include lower and absolute, not relative gene copy number. The data has high precision and has better tolerance to inhibitors. These characteristics can lead to better and more precise molecular tests in food. , Before dPCR wide spread adoption is seen, however, the limitations of high cost and limited dynamic range must be addressed.

It’s not only in the testing labs and adjacent technologies that NGS is having an impact. In the labs driving innovation in food and food ingredient development, applications of NGS are being used to develop targeted food ingredients.

Nestle is the leader in this convergence of food, health and nutrition and over the last three years, the company has acquired and formed partnerships targeting the space. In its formation of the Nestle Institute of Health Sciences, Emmanuel Baetge, head of NHIS, emphasized NHIS expertise and research capabilities using systems biology, next generation sequencing, and human genetics.

The world of food safety is as dynamic as the natural flora of food itself. Changing regulations, evolving organisms, technological change and consumers’ changing tastes require new solutions. The requirements of the food laboratory have not changed. They are the protectors of brands and the teams we trust to deliver safe and quality foods. However, how they do that has and will continue to change.

Next time… molecular serotyping.


  1. Wetterstrand KA. DNA Sequencing Costs: Data from the NHGRI Genome Sequencing Program (GSP) Available at: www.genome.gov/sequencingcosts. Accessed 1/13/2014 [DOA 1/13/12014].
  2. Beilei Ge and Jianghong Meng , 2009 14: 235 Advanced Technologies for Pathogen and Toxin Detection in Foods: Current Applications and Future Journal of Laboratory Automation DOI: 10.1016/j.jala.2008.12.012.
  3. Morisset D, Sˇ tebih D, Milavec M, Gruden K, Zˇ el J (2013) Quantitative Analysis of Food and Feed Samples with Droplet Digital PCR. PLoS ONE 8(5):e62583. doi:10.1371/journal.pone.0062583.
  4. http://www.nestle-nespresso.com/asset-libraries/Related%20documents%20not%20indexed/Nespresso%20poster%20ASIC2012%20DNA%20traceability.pdf
Dan Okenu, Ph.D., Food Safety Manager, H-E-B
Retail Food Safety Forum

Testing and Evaluation of Food Safety Tools Simplified

By Dan Okenu, Ph.D.
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Dan Okenu, Ph.D., Food Safety Manager, H-E-B

The management of Food Safety and Quality Assurance (FSQA) system is a key business function that plays a very important role in the sustenance of the food industry. Its primary objective is to produce and serve safe quality food to consumers, through compliance with all relevant Federal, State and Local regulatory laws. It assists in the reduction of food wastage or food spoilage, and thus has a strong impact on the bottom line. Proper management of the FSQA system protects business brands, ensuring that they don’t become part of the gloomy statistics on foodborne disease outbreaks and damaging recalls. In all cases, the protection of the entire public health remains sacrosanct, and in fact, closely aligns with the primary business objective of getting a reasonable return-on-investment. Inevitably, businesses rely on a timely and cost-effective project management to ensure that their FSQA system remains relevant and sustainable for a continuous business growth.

Potential sources of new FSQA projects

Projects intended for the improvement of an FSQA system may be identified and initiated based on input from the following sources:

  • Regulatory compliance with applicable Federal, State and Local laws;
  • Voice of customers through complaints obtained by customer calls;
  • Technology-driven continuous improvement to upgrade to a smarter method, process, equipment or service;
  • Voice of business franchise operators, owners, managers and team members aimed at improving operational efficiency; and
  • Operational challenges observed by corporate staff during field visits.

Examples of FSQA projects that require testing and evaluation

At every stage, there will be tons of very important projects requiring urgent attention and competing for limited resources with corporate advertising and brand campaigns which have fixed budgets. Some of these projects may be as simple as putting a new dish-washing scrub pad in the system. This project may have been initiated following several reports by team members that current green scrub pad is not effective and also releases greenish color with scrub pad debris reported in ready-to-eat (RTE) foods. The associated risk is that foreign material in food constitutes a health hazard while improper washing of dish wares may lead to cross-contamination and outbreak of foodborne illness. This is easy but still requires testing to confirm that the new scrub pad is the best cost-effective option. Other projects however may be as complex as introducing a new produce (fruits & vegetables) safety system that includes a pathogen kill-step, instead of the regular cold water rinse. This will provide an extra layer of produce safety at the retail level, in case the system fails at the processing plant facility level, for instance, in the case ofthe multistate Listeria outbreak involving Cantaloupes from Jensen Farms, Colorado in 2011.

Another new initiative could be working with suppliers to validate a new method for detecting bone fragments and physical contamination of boneless poultry meat. Revamping the automated dish-washing room to improve food code compliance is a multifaceted project that requires a lot of resources and planning for a successful testing and evaluation.

Testing and evaluation milestones

A systematic approach is required to properly test and evaluate new FSQA products or services before a chain-wide roll-out is authorized by management. Depending on whether we are looking to introduce a new product or service into the system, some of the testing and evaluation milestones may include a combination of:

  • R&D to determine and evaluate options to resolve issue;
  • Review of options for industry best practices by FSQA team;
  • Cross-functional team evaluation by stakeholders to determine impact on key business functions, including a robust business analysis to determine cost implications;
  • Vendor verification to certify compliant business status;
  • Execution of a Non-Disclosure Agreement (NDA) or Master Vendor Agreement (MVA) between corporate and vendor partner, to legally protect all parties;
  • Preliminary testing at the corporate Technical Center for proof of principle and to evaluate product safety and potential OSHA requirements at a controlled environment;
  • One store test to determine operational feasibility in an actual business environment;
  • Three to 10-store testing to evaluate operational dynamics in a larger number of stores;
  • Thirty to 60 store-market testing in different markets to carefully monitor usage and operational outcome and ensure compliance and expectations, and extrapolate results to mimic a national chain wide roll-out;
  • Performance survey of test stores, data collation, analysis and review of results, followed by management approval;
  • Chain-wide roll-out by a cross functional team representing all impacted areas of the business; and
  • Post-chain-wide roll-out follow-up to monitor usage and resolve any lingering issue, namely:
  • Adequate SOP training to reinforce proper use;
  • Vigorous marketing campaign to increase chain-wide usage and compliance;
  • Effective ordering and delivery logistics; and
  • Potential short term and long term quality issues.

Developing an FSQA project matrix template

Certainly, the need to initiate new FSQA projects will increase as the various parts of the new Food Safety Modernization Act (FSMA) come into full force. Since FSMA brings a new regulatory burden on the food industry, its full implementation will require new ways of doing business, and most likely will affect the overall cost. This calls for a smarter management strategy to keep costs down and customers happy. The massive number of legitimate but competing food safety projects literally begging for attention can be overwhelming for Managers, especially with resources always in short supply. With this scenario, it is critical for the Manager to develop an FSQA project matrix template that delineates the level of importance of each project based on overall risk assessment, cost-benefit ratio, regulatory food code requirements, and buy-in by stakeholders, including management and final end-users.

FSQA project implementation and recipe for success

Testing and evaluation can be an expensive venture considering the test duration, number of test stores involved and capacity utilization for test products. The good news however is that most vendors are willing to fund substantial portion or even the entire test. This is essentially because vendors want to demonstrate that their product works, are in compliance, certified and approved by relevant federal, state and local agencies, and fulfills all obligations as outlined in the statement of work. It is a win-win situation for both vendor and corporate because once a product is approved for chain wide roll-out, it can stay in use for several years until an upgraded becomes available. Thus, corporate funding commitment may be minimal and restricted only to staff time for overseeing the testing process. It is important to mention that training is a critical component at every stage of the testing process. Standard operating procedures, training video clips and on-site training are required to ensure that test product is used according to manufacturer’s instruction and in compliance with all relevant regulations. Due diligence and proper training of end-users including store managers, team members and associates will ensure that the roll-out of a food safety tool to mitigate an existing risk does not introduce a new risk in food service operations. An example is the introduction of a new disinfectant to comply with a new regulation that requires a Norovirus approved disinfectant grade chemical for cleaning playgrounds. The disinfectant however is not approved for food contact surfaces since it’s not a regular strength sanitizer.

Consequently, any inadvertent cross-usage on food contact surfaces may constitute a serious food safety risk. Similarly, an SOP training gap may result in higher risk if associates using yellow color-coded aprons for raw food processing cross-contaminates the RTE food board areas with raw chicken/beef contaminated aprons. For instance, the Costco Rotisserie Chicken recall of late 2013 appears to have been linked with Salmonella cross-contamination after the cooking process in the food preparation area. Thus, proper training on the useof food safety tools and processes is critical both during product testing and post-chain-wide roll-out, to accurately evaluate and monitor risk mitigation practices.

To enable success, food retail chains employ the services of third party consultants to assist in-house staff and bring project-specific subject matter expertise to the table for rigorous risk assessment and risk mitigation. This strategy will also assist in timely communication that support buy-in by senior management and other relevant stakeholders. In addition, the implementation of such projects will remain effective and efficient, freeing up valuable time for corporate staff to continue supporting the business in the most critical areas of providing seamless customer services. Most importantly, a third party working in concert with vendors and corporate staff will bring an unbiased product testing and evaluation standard that cannot be left entirely at the discretion of vendor partners.

Proper documentation is required at every stage to ensure that all potential confounding factors are considered and evaluated at every level. Surveys, feedback compilation and analysis by a third party will assist in building credibility for test data, and enable management have the right set of data to make an informed business decision. Some level of customization may be involved as issues raised by stakeholders are addressed during the testing process. Open communication is important to keep all parties in the loop and encourage honest discussion of issues and how best to resolve them in a cost-effective manner.


Testing of new FSQA tools and services is a cost saving process that helps Managers to completely resolve potential issues upfront before introducing products into the system. Improperly tested food safety products may lead to a breach in the system down the road. Ordinarily, the use of transparent plastic wrap to cover raw chicken pans during the thawing process is an excellent barrier against cross-contamination of food-contact and non-food contact surfaces with raw chicken juice. However, the transparent nature of the plastic wrap makes it extremely difficult to see a torn piece of plastic wrap inside the raw chicken pan. Due diligence during testing should identify such aberration and resolve it by customizing into easily identifiable yellow color coded plastic wrap. This test-mode corrective action will ensure that torn pieces of plastic wrap won’t get into food served to customers after chain-wide roll-out.

While proper testing, evaluation and roll-out of new FSQA products and services may be laborious, time consuming and somewhat expensive, it is still considered one of the industry best practices that supports the delivery of safe quality food to customers and protects the business brand. Overall, it benefits businesses in the long run to budget enough resources for this very important business function, instead of postponing or scrapping risk mitigation programs until crisis situation that may hurt customers, business brand and undermine return-on-investment.