Quantitative genetic analysis plays a critical role in the research and development of novel biotherapeutics. As a sensitive and precise method for quantitating specific nucleic acid targets, qPCR is used broadly by researchers for applications including gene expression analysis, microbial detection, single nucleotide polymorphism (SNP) genotyping, and more.

For experiments that aim to detect or quantify multiple targets, two or more probe-based qPCR assays can be combined in a single reaction well. However, developing a multiplex qPCR experiment is more complex than adding an extra assay to the reaction mix. To set a multiplex qPCR experiment up for success, it is important to consider the technical requirements and complexities of multiplexing.

In this webinar, real-time PCR expert and Senior Field Application Scientist Dr. John B. Pfeifer draws on his 30+ years of industry experience to present strategies for implementing robust and reliable multiplex qPCR analysis. Join Dr. Pfeifer as he sets the stage with key foundational concepts before walking you through the technical considerations, benefits, and challenges of multiplex qPCR. Following the presentation, stay tuned for a Q&A with Dr. Pfeifer and get answers to your multiplex qPCR questions.

Learning objectives:

1. Review fundamental concepts of real-time PCR and multiplexing.

2. Learn how multiplex qPCR analysis can increase throughput, reduce reagent & sample consumption, and improve precision.

3. Explore common multiplex scenarios to understand the technical considerations for multiplex study designs.

4. Understand how to perform multiplex assay verification and the complexities associated with verifying multiplexed assays with 3 or more targets.

We will discuss the Digital LightCycler System, a digital PCR (dPCR) system for DNA and RNA with 6-color capability for flexible, multiplexed biomarker detection.  With advances in sensitivity, precision, flexibility, and integration, the Digital LightCycler System is a powerful digital PCR system that aims to help laboratories push the boundaries of clinical research forward.  Three configurations of consumables are available to meet varied application needs, including high sample volume for cfDNA screening or greater number of partitions for high-resolution CNV analysis.  We will present data that demonstrates the sensitive performance of the Digital LightCycler System as the powerful new addition to the Roche PCR ecosystem.

Molecular diagnostic assay development and design is no simple task, and several factors affect the performance and usability of a test. Diagnostic companies can enhance assay stability and consistency, while reducing supply chain and shipping costs, by developing freeze-dried (lyophilized) assays. Because of these benefits, lyophilization can also enable accelerated entrance into global markets, as well as field use due to the ability to ship and store at ambient temperatures.

One major challenge in assay development and lyophilization is incompatibility with conventional enzyme formats. It is imperative that enzymes used do not contain glycerol in their buffers, as this interferes with the freeze-dry process. Lyo-ready enzymatic solutions must offer equal sensitivity, specificity, and reproducibility to ensure a robust assay.

Another hurdle in assay lyophilization is the development of an in-house freeze-dry process. In order to do this properly, companies must spend significant amounts of money on capital equipment, headcount, and infrastructure (humidity-controlled rooms, liquid nitrogen set-up, etc.). In this webinar we will discuss some of these challenges, as well as options for outsourcing lyophilization.

Learning Objectives: 

1. Discover the many apparent (and hidden) benefits of molecular diagnostic assay lyophilization, as well as important considerations in process development. 

2. Review how the challenges of assay lyophilization can be mitigated by incorporating enzymes with low glycerol content that still perform comparably to standard formats.

3. Summarize the important considerations in developing a freeze-dry process.

Access to and experiences with healthcare continue to be inequitable worldwide even at the level of clinical laboratory testing. From building and sustaining a thriving, diverse workforce to adopting more inclusive testing practices to best serve an increasingly diverse patient population, there are many ways the clinical lab can also contribute to reducing health disparities amongst vulnerable communities. Here we present an overview of various initiatives and achievements within the Department of Laboratory Medicine and Pathology at Mayo Clinic to transform praxis through the power of equity, inclusion, and diversity. We demonstrate that adopting measures to support, empower, and educate one another on diverse perspectives, establish a culture of safety and belonging, create opportunities for historically disenfranchised populations, and broaden our analytical, interpretive, reporting, and research efforts to be more inclusive of all individuals can not only improve laboratory science but also positively impact patient outcomes.

Learning Objectives: 

1. Describe measures for enhancing gender inclusivity in clinical testing.

2. Explain how race and ancestry can impact test results and representation in research. 

3. Discuss strategies for improving equity, inclusion, and diversity in the laboratory workforce.  

Lateral flow assays (LFA) are simple diagnostic tools that can be used to detect a variety of biomolecules that include biomarkers, pathogens, and environmental contaminants. Many industries use LFAs as they are typically rapid assay devices that are especially useful as point-of-care diagnostic devices and as a public health tool that can be sold to users directly. Sartorius is globally recognized a as leading supplier of biotherapeutics and in vitro diagnostics (IVD) solutions.

Antibodies play a crucial role in the development of LFAs. The quality attributes of these antibodies including the understanding of how they interact with target antigens are essential for the development and optimization of LFAs. The Octet® system is a powerful label-free and easy-to-use high-throughput solution for analyzing biomolecular interactions. In the product development of LFAs, the Octet® system, a Sartorius solution, can be used to investigate antibody-antigen interactions, to perform assay development and optimization and as a reagents quality control tool. 

As an attendee, you will learn more about: 

• The assay design and considerations in the development of lateral flow assays.

• The various applications of the Octet® label free systems for biomolecular interactions.Learn of the various applications of the Octet® label free systems for biomolecular interactions.

• How Octet® systems can be used as quality control platforms for the selection of optimal antibodies for the development of lateral flow assays.

Medical laboratories are an integral part of the healthcare system. They supply crucial diagnostic data that underpins disease detection, patient care, and public health management. The quality of lab results—which encompasses the accuracy, reliability, and timeliness of reported test results—is of utmost importance in ensuring accurate diagnoses, reliable treatment decisions, and timely patient care. Poor quality in lab processes can lead to severe repercussions, including unnecessary treatments, delayed diagnoses, treatment complications, and increased costs. Therefore, it is imperative to establish a robust quality management system (QMS) to mitigate these risks and deliver high-quality lab services.

Recognizing the importance of quality management in lab operations, global standards such as ISO 15189 have been developed to adapt the QMS framework for medical labs. ISO 15189 is a globally recognized quality standard specifically designed for medical laboratories. Since its initial release in 2007 and its most recent update in 2022, ISO 15189 has played a significant role in ensuring the competence, quality management, and technical processes within medical laboratories. ISO 15189 focuses on the unique needs of clinical diagnostic labs and encompasses a broad range of areas, including QMS, personnel proficiency, facilities, pre-analytical, analytical, and post-analytical procedures, document management, and customer satisfaction. By adhering to ISO 15189, medical laboratories can enjoy numerous benefits, including enhanced accuracy and reliability of test results, improved patient safety, heightened confidence in lab services among healthcare providers and patients, and standardization of lab practices. ISO 15189 also enables labs to meet regulatory requirements and facilitates international recognition and accreditation. The standard promotes a culture of continuous improvement and top-notch customer services within medical labs.

Learning objectives:

1. Define the Importance of ISO 15189 accreditation for clinical diagnostic labs.

2. Explain the various requirements of ISO 15189 compliance and the notable updates in ISO 15189:2022 as compared to the previous version, ISO 15189:2012.

3. How a LIMS can help clinical diagnostic labs achieve ISO 15189 accreditation, demonstrate competence, and assure quality testing.

Immunoassays and mass spectrometric assays are the two major methodologies in contemporary clinical chemistry laboratories. While immunoassays are highly automated and offer a faster turnaround time, mass spectrometry has several advantages over immunoassays. As mass spectrometry continues to expand in clinical chemistry, it is widely used for the measurement of small molecules, including drugs in urine, therapeutic drug monitoring, and hormones. However, the adoption of this technology for targeted protein analysis has been slower. This presentation will describe the key components of targeted protein quantitation using mass spectrometry, introduce recent advancements in clinical applications, and present available resources to enhance its efficiency for clinical laboratories.

Learning Objectives:

1. Explain the advantages of mass spectrometry in protein analysis.

2. Describe the workflow for targeted protein quantitation.

3. Apply the concepts to evaluate clinical protein assays.

This talk will describe the role of the clinical lab in advancing health equity. It will define the gaps in equitable access to lab testing and how that impacts patient care. We will also cover how equitable lab testing can be achieved through novel assay development, clinical test development, regulatory approval, and clinical test implementation. The goal of this talk is to inspire laboratorians to get involved in advancing health equity.

Learning Objectives: 

1. Understand how lab testing can contribute to health care inequity.

2. Describe how equitable access to lab testing can positively impact patient care.

3. Detail the numerous ways that laboratorians can be involved in lab equity.

Despite an overall good performance on all laboratory procedures used to detect monoclonal (M) proteins or paraproteins, mis- or false-diagnoses have been seen using electrophoresis methods, especially on cases with non-classic abnormal immunoglobulin profiles. This session will highlight cases with these challenging profiles and present solutions using multi-method approach or mass spectrometry to address potential misdiagnosis. In addition, monoclonal antibodies (mAb) used as immune-modulatory therapy have been emerging for the treatment of cancers including multiple myeloma and autoimmune diseases. Clinical laboratorians and pathologists should be aware of the interference from mAb therapeutics on M protein assessment. In this session, we will present a comprehensive overview of laboratory methods used for paraproteinemia analysis and ways to identify/eliminate interferences associated with these methods. Mass spectrometry-based analysis (MASS-FIX assay) of M proteins provides another option in investigation of abnormal cases. The advantage of mass spectrometry-based method over conventional electrophoresis will also be discussed.

Learning Objectives:

1. Summarize the advantages and limitations of various methodologies used for paraproteinemia analysis.

2. Review the advantage of using mass spectrometry-based method over conventional electrophoresis method on abnormal paraproteinemia cases.

3. Develop knowledge on latest technologies and approaches used to eliminate interferences on M protein analysis from mAb therapeutics.

Traumatic brain injury (TBI) is a widespread global health concern, impacting approximately 10 million individuals annually. Beyond the initial acute phase, TBI often results in enduring neurological sequelae. Current diagnostic approaches, including electroencephalogram, computed tomography, and clinical assessments like the Glasgow Coma Score fall short in providing timely and precise TBI characterization. This presentation offers an overview of emerging non-invasive blood-based markers for TBI, encompassing proteins, nucleic acids, and extracellular vesicle-associated markers, with implications of emerging technologies including near-patient testing devices. These advancements hold significant implications for the improved management of TBI, potentially enabling early intervention and more tailored treatment strategies.

Learning Objectives:

1. Review the global impact of traumatic brain injury (TBI) and its long-term neurological consequences.

2. Explore the limitations of current TBI diagnostic methods in accurately characterizing and timely identifying TBI.

3. Gain insight into the promising field of non-invasive blood-based markers for TBI, including proteins, nucleic acids, and extracellular vesicle-associated markers.

Interference in laboratory assays is a concern since it can positively or negatively influence test results, which can affect patient care. Interferents could be endogenous or exogenous compounds. Dietary supplements such as biotin and vitamin C are examples of exogenous interferents in common laboratory assays. During this presentation, mechanisms underlying such interference will be discussed, along with ways of detecting it and possible solutions.

Learning Objectives:

1. Outline an overview of analytical interferences in laboratory assays.

2. Describe potential interference caused by megadose biotin and vitamin C supplementation.

3. Discuss strategies to detect and mitigate such interferences.