Flow cytometry is crucial for diagnosing hematological malignancies. However, the rising number of tests and ongoing labor shortages are placing significant pressure on both the lab and its staff. Lab automation, especially automating data analysis in flow cytometry, can improve efficiency, scalability, and reduce costs. In today's talk, I will begin by introducing the concepts behind flow cytometry data analysis, the strategies for automation, and the essential tools and modules involved. I will then showcase two of our projects to illustrate how we have developed machine-aided analysis systems tailored to clinical and research needs. Finally, I will conclude by discussing our lab's future development plans.

Learning Objectives:

• Strategies to automate flow cytometry data Analysis in clinical setting

• Modules and techniques in analyzing flow cytometry data

• Strategies in the induction treatment scenario,
• Even to delineate candidate biomarkers for further enrichment and patient stratification can be explored.

• Outline the total spectrum of evolving systemic treatment options in non-small cell lung cancer
• Identify the impact of immunotherapy and molecular targeted treatments on patient outcome and the potential relevance of circulating tumor (ct)DNA assessment on guiding treatment
• Discuss the integration of systemic treatment options in the multimodality setting of nonmetastatic locoregional disease

Join this year's poster presenters in the Poster Hall during the Poster Discussion, Wednesday, November 13th, from 10:00am – 11:00am PST, to chat live about their posters and latest research and findings.

If you use a pipette on a daily basis, then this webinar is for you. Pipettes are used to prepare samples, blanks, and standards for a range of analytical experiments. This is why the wrong type of pipette, improper technique, or a poorly maintained pipette can have a negative impact in your data. 

In this webinar we will talk about how different pipetting techniques can influence your pipetting results and review tips and tricks on choosing the right pipettes and adjusting your technique for different liquids and conditions. Finally, we will go through the importance of regular pipette maintenance and calibration to ensure reliability of results and conformance to international standard ISO8655.  

Learning Objectives:

1. Explain why pipetting technique matters.

2. Discuss how to pipette challenging liquids.

3. Analyze the importance of regular maintenance and calibration.

Pharmacogenomic testing uses a patient’s genome to identify their response to a drug, forming a key part of personalized medicine. Hundreds of clinical pharmacogenomics (PGx) tests are currently available on the market in the United States. With so many tests available, how can you determine that a test is clinically and analytically valid and holds clinical utility?

In this webinar, our expert speaker will outline the ACCE Model Project Framework as a tool for evaluating pharmacogenomic tests, describing the key considerations to take into account when implementing these tests. You’ll learn how to select the genetic variants to be tested, choose the appropriate genotyping technology and develop laboratory procedures for sample processing and data analysis.

Learning Objectives

• Explain the ACCE Model Project Framework for evaluating genetic tests, specific to PGx tests
• Define analytical and clinical validity and clinical utility and their importance to pharmacogenetic testing stakeholders
• Discuss key considerations laboratorians need to ponder when implementing a PGx test that meets ACCE standards

Join us to explore recent advancements and applications of isothermal amplification techniques, such as LAMP, RCA, and RPA. We will compare these methods with traditional PCR, discuss their benefits for point-of-care diagnostics, and present real-world examples, including detection of dengue virus and equine pathogens. Additionally, we will address challenges in implementation and highlight innovations driving future developments in this field. This webinar aims to provide a comprehensive understanding of isothermal amplification and its potential to revolutionize pathogen detection at the point of care.

As we advance through 2024, significant updates in Human Immunodeficiency Virus (HIV) and Sexually Transmitted Infections (STI) prevention and treatment are emerging. Recent clinical trials and global research highlight both advances and ongoing challenges in these critical public health areas. How will these developments influence current practices and shape future strategies in HIV and STI management?

In this webinar, our expert speaker will explore these updates, presenting recent epidemiological data on HIV and STIs worldwide to provide a clear picture of current trends. You’ll gain insights into the latest advancements in HIV treatment and prevention, including innovations in long-acting formulations. The session will also cover recent progress in STI treatment and prevention, focusing on emerging strategies such as the use of doxycycline as postexposure prophylaxis.

Learning Objectives:

• Understand recent epidemiology of HIV and STIs globally
• Learn updates on HIV treatment and prevention, including long-acting formulations
• Learn updates on STI treatment and prevention, including doxycycline as postexposure prophylaxis

This webinar is designed to provide participants with an in-depth understanding of the latest advancements in lyophilization-ready (Lyo-ready) products tailored for assay development. It will cover the critical aspects of product customization, the benefits of 1-step RT-qPCR systems, and the capabilities of Thermo Fisher Scientific in providing custom commercial supplies. Attendees will gain valuable insights into optimizing assay performance and reliability through innovative Lyo-ready solutions.

Learning Objectives:

• Understand the fundamental principles and benefits of lyophilization-ready products in assay development, including stability and ease of use.
• Explore the customization capabilities offered by Thermo Fisher Scientific, enabling tailored solutions to meet specific research and commercial needs.
• Gain practical knowledge on the application and advantages of the 1-step RT-qPCR system in enhancing assay accuracy and efficiency.

Interpretation of human herpesvirus type 6 (HHV6) detection in the cerebrospinal fluid (CSF) of children can be complex; the virus can cause acute infection, reactivation, or can be inherited chromosomally integrated (iciHHV6). Here we will describe HHV6 virus, the epidemiology and clinical presentation of HHV6 disease, review evidence of clinical consequences of inherited chromosomally integrated HHV6 and summarize our study on detection of HHV6 in pediatric CSF samples.

Learning Objectives:

• Describe HHV6 virus, the epidemiology and clinical presentation of HHV6 disease 

• Review evidence of clinical consequences of inherited chromosomally integrated HHV6 

• Summarize our study on detection of HHV6 in pediatric CSF samples 

Prime editing has gained recognition as a highly precise genome editing method, often outperforming other Cas9-based techniques. We recently introduced innovative nCas9-DNA polymerase fusion proteins (DOI: 0.1101/2024.07.09.602710), which create chimeric oligonucleotide-directed editing (CODE) systems designed for targeted "search-and-replace" genome modifications. After extensive engineering with testing over 26 purified variants, we developed two efficient editors, CODEMax and CODEMax(exo+), that achieve accurate genome edits in human cells with minimal off-target effects. Both systems incorporate a modified Bst DNA polymerase with strong strand displacement activity, while CODEMax(exo+) also includes a 5’ to 3’ exonuclease function to facilitate more effective strand invasion and repair, ensuring the desired edits are successfully integrated. Our results show that CODEs perform small insertions, deletions, and substitutions with greater accuracy compared to PEMax across various loci. Ultimately, these CODE systems complement existing prime editors, expanding the toolkit for genome manipulation without inducing double-stranded breaks. 

Beyond CODE, we are engineering a range of CRISPR-based tools to not only improve precise genomic surgeries in vivo but also to advance clinical diagnostics. Some of these technologies like PICNIC (DOI: 10.1101/2024.05.02.24306194) can expand the range of sequences that can be targeted in the genome from ~1% to ~100% by eliminating the need for a PAM.

Learning Objectives:

• Recognize the need for a simple and efficient gene correction technology

• Identify the limitations of existing genome editing technologies in terms of efficiency and off-target effects.

• Describe how CRISPR/Cas systems can be coupled with engineered DNA polymerases to achieve precise modification of any genomic target.

Accurate glycemic-monitoring and control of the patients with hyperglycemia/diabetes are essential for better patient care management. Hemoglobin A1c (HbA1c) measurements correlate with the average plasma glucose level over the preceding 2-3 months. HbA1c is a stable, glycated hemoglobin formed by non-enzymatic addition of glucose to amino-terminal end of Hb polypeptide chains. HbA1c is stable through the life of red blood cells (RBCs) and can be measured regardless of the time of the day and fasting status. Therefore, HbA1c testing has become the standard of practice for glycemic control assessment. According to the American Diabetes Association (ADA) guidelines, through National Glycohemoglobin Standardization Program (NGSP)-certified Diabetes Control and Complications Trial (DCCT)-standardized assay, an HbA1c level between 5.7% and 6.4% is defined as prediabetes, and a level of 6.5% or more is consistent for diagnosis of diabetes. Hemoglobin (Hb) variants arise due to mutations causing changes in Hb alpha or beta chain aminoacid at a particular sequence. Many methodologies are available for HbA1c testing some of which can affect the results based on Hb variant. In this session, principles of HbA1c assays such as ion-exchange high-performance liquid chromatography (HPLC), boronate affinity HPLC, capillary electrophoresis, immunoassay and enzymatic assays will be explained. The analytical and biological limitations of these assays-such as hemoglobinopathies (Hb variants) will be discussed and compared. In addition, details surrounding CAP proficiency testing (PT) limits, CLIA rule and the efforts of NGSP related to HbA1c testing improvement will be presented.

Learning Objectives:

• Understand the background of diabetes and importance of HbA1c for diabetes monitoring
• Review various methodologies and best practices for HbA1c Testing
• Evaluate role of NGSP and CAP Proficiency Testing Program for HbA1c

Background
The purpose of the study is to evaluate the performance of the urinalysis (UA) screening analyzers. Rapid point-of-care (POCT) urine dipstick testing in the emergency department (ED) was compared to a laboratory automated platform.

Methods
A total of 30 urine specimens were collected from the emergency department at our hospital and were simultaneously tested for urinalysis (UA) on POCT Siemens Clinitek Status+ analyzer using the Multistix 10SG strip. In the laboratory we performed the tests on the automated ARKRAY Aution MAXTM instrument, using the Aution Sticks 9EB strips. Samples with more than a trace of leukocyte esterase and/or nitrates are considered positive for a urinalysis and further, the samples were reflexed to urine culture. The performance of both platforms was analyzed using EP Evaluator (Data Innovations LLC.).

Results
A total of 30 urine specimens were analyzed for UA using both the POCT and automated laboratory analyzers. The agreement (95% CI) between leukocyte esterase, nitrites, and blood are as follows: 53.3% (36.1 to 69.8%); 96.7% (83.3 to 99.4%), and 70.0% (52.1 to 83.3%) (see Table 1.). Furthermore, out of the 30 specimens that were tested on both platforms, there was prominent discordance between the POCT and lab UA test. The 7 urine samples that were negative on POCT were positive on the laboratory UA platform. All 7 were sent for urine culture, out of 7 only 1 sample was positive for culture which was abnormal: >100,000 CFU/ml Escherichia coli. Out of 30 specimens that were tested on both platforms, 23 were in concordance with both the platforms out of which 8 patients tested positive for culture (5 were E. coli, 1 was E. coli combined with Klebsiella pneumoniae, 1 was solely Klebsiella pneumoniae, 1 was Staphylococcus saprophyticus, and 1 was Streptococcus agalactiae). The remaining 15 patients resulted for no significant growth of bacteria or suggesting contamination. The sensitivity of POCT UA and laboratory-performed UA was 89% and 100% respectively. Furthermore, the specificity of POCT UA and laboratory-performed UA was 36% and 7% respectively. 

Conclusion
Our data shows although POCT UA is rapid, a prominent disparity is observed between the POCT UA and automated lab testing. The lab-performed UA has greater sensitivity and specificity compared to the POCT UA. Clinicians should interpret the urine dipstick test results in a clinical context that includes the patient’s history and symptoms, especially when using POCT in the ED. 

Learning Objectives:

• Comparing the performance of  the two urinalysis platforms

• Identifying the gaps between the two platforms

• Recommend education on disconnect between patient’s symptoms vs test results.

Cardiometabolic syndrome is a combination of metabolic dysfunction mainly characterized by insulin resistance, impaired glucose metabolism, dyslipidemia, and hypertension. Cardiac remodeling, coronary heart disease, and even heart failure could result from metabolic syndrome. Currently, around 25% of the adults worldwide are suffering from this syndrome. Along with the increased rates of obesity, diabetes and hypertension in the past decades, there has been an increase in the incidence of CMS. With these standard risk factors, the scientific research aims to focus on addressing the diverse constellation of risk factors and identifying new diagnostic biomarkers that incorporates an integrated approach to treating these risk factors across diverse populations. In this session, the pathophysiology, risk assessment for precision health, molecular and metabolic biomarkers of metabolic cardiovascular disease, especially posttranslational modification of troponin will be presented and discussed.

Learning Objectives:

• Pathophysiology of cardiovascular consequence of metabolic syndrome
• Molecular insights in metabolic cardiovascular disease
• Risk assessment, metabolic biomarkers and the evolution of proteomic troponin analysis for predicting CMS

Hemolysis is a common pre-analytical variable that can lead to inaccurate results, specimen rejection, and specimen recollection in the clinical laboratory. Hemolysis is most often an in vitro phenomenon, usually occurring due to specimen handling. In this presentation, we will review common pre-analytical causes of hemolysis and how hemolysis impacts different clinical chemistry analytes. The presentation will conclude by discussing Special Handling Instructions (SHI), a sample handling process that was developed to reduce hemolysis in samples collected from hospitalized patients with repeatedly hemolyzed specimens. Data from a study that evaluated the effectiveness of SHI to reduce hemolysis will be shared.

Learning Objectives:

• Describe common preanalytical causes of hemolysis
• Describe how and why hemolysis interferes with the measurement of different clinical chemistry analytes
• Review strategies to reduce hemolysis in repeatedly hemolyzed specimens

Stable isotope-labeled internal standards are widely used in laboratory-developed clinical mass spectrometry tests. An equal amount of these internal standards is added to all samples in a batch, including calibrators, controls, and unknowns. The ratio of the analyte signal to the internal standard signal is then used to create the calibration curve for quantification. Proper use of internal standards can normalize variations in recovery throughout the entire analytical process, which includes sample preparation, injection, chromatography, and mass spectrometry ionization. Key criteria for selecting an internal standard for a small molecule analyte typically include: 1) an identical chemical structure except for the isotopes or at least a similar structure, 2) chemical stability, 3) a mass-to-charge ratio difference that can be easily distinguished by mass spectrometry, and 4) absence from the sample matrix. Despite following these general guidelines, pitfalls may still arise, and improper use of internal standards can lead to biased clinical results. In this presentation, two troubleshooting cases will be discussed as part of our laboratory's quality improvement projects for existing assays. The first case addresses a non-linear response issue in the urine 5-Hydroxyindoleacetic acid (5-HIAA) test, while the second addresses high imprecision issues in the Sirolimus test at low concentrations. Both cases were resolved by switching to more suitable internal standards, resulting in improved accuracy, linearity, and precision in the optimized assays.

Leaning Objectives:

• Understand the role of internal standards in isotope dilution LC-MS/MS tests
• Identify general criteria for selecting an internal standard in laboratory-developed LC-MS/MS tests
• Learn from two troubleshooting cases how improper use of internal standards can lead to biased results

Clinical laboratory analyzers and automation systems have a limited lifespan, and timely upgrades are essential to maintaining reliable, cutting-edge technology that supports optimal patient care. A request for proposal or request for information (RFP/RFI) provides an ideal opportunity for clinical laboratories to evaluate and compare vendor options that can address existing challenges while preparing for future demands. In this presentation, the speaker will introduce the RFP/RFI process and highlight key factors to consider when upgrading instrumentation in a clinical laboratory.

Learning Objectives:

• Summarize the RFP/RFI process
• Discuss key elements to consider when changing instruments in a clinical laboratory

Over the past decade, the opioid crisis in the United States has been exacerbated by the fentanyl epidemic. Fentanyl, a synthetic opioid, is approximately 100 times more potent than morphine and 50 times more potent than heroin. Due to its high potency, detecting fentanyl use has become increasingly critical. Laboratory testing for fentanyl provides essential information in various settings, including acute intoxications or overdoses in emergency centers, compliance monitoring for pain management, addiction treatment selection for substance-use disorders, and behavioral testing programs.
While mass spectrometry-based methods are considered the gold standard for confirmation, urine testing using immunoassays remains the most common and feasible screening method in many institutions. Several FDA-approved immunoassays are available for automated chemistry analyzers, targeting either fentanyl or its metabolite norfentanyl, with varying degrees of cross-reactivity with other fentany analogs. Rapid testing methods, such as lateral flow immunoassays, have also gained interest for fentanyl screening in specific clinical settings.
This presentation aims to review recent publications on the analytical and clinical performance of various fentanyl immunoassays and share our experience in evaluating these assays for cinical implementation.

Learning Objectives:

• List currently available fentany! immunoassays
• Describe the advantages and limitations of different fentanyl testing methodologies
• Evaluate fentanyl immunoassays for implementation based on clinical need