No More Dead Ends: Improving Legionella Testing with Viability qPCR

Image of cooling towers.

Legionella is the causative agent of Legionnaires’ disease, a severe form of pneumonia with a mortality rate of around 10%​. Contaminated water systems, including cooling towers and hot water systems, serve as primary reservoirs for this opportunistic pathogen. Traditional plate culture methods remain the regulatory standard for monitoring Legionella, but these methods are slow—often requiring 7–10 days for results—and suffer from overgrowth by non-Legionella bacteria​. Additionally, traditional methods fail to detect viable but non-culturable (VBNC) bacteria—cells that remain infectious but do not grow on standard culture media. 

Molecular methods like PCR-based detection provide faster and more sensitive Legionella identification. However, a key limitation persists: PCR detects DNA from both live and dead bacteria, leading to false positives and unnecessary or even wasteful remediation efforts​. To address this challenge, Promega has developed a viability qPCR method that retains the speed of molecular testing while distinguishing viable bacteria from non-viable remnants. In this third blog in our Legionella blog series, we cover how molecular detection methods can be refined to provide actionable results for Legionella monitoring. 

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Overcoming qPCR Inhibitors: Strategies for Reliable Quantification 

Today’s blog is written by guest blogger, Gabriela Saldanha, Senior Product Marketing Manager at Promega.

Quantitative PCR (qPCR) is an indispensable tool for nucleic acid analysis, widely used in research, clinical diagnostics and applied sciences. Its sensitivity and specificity make it a powerful method for detecting and quantifying DNA and RNA targets. However, qPCR reactions are highly susceptible to inhibitors—substances that interfere with enzyme activity, primer binding, or fluorescent signal detection. These inhibitors can originate from biological samples, environmental contaminants, or laboratory reagents, potentially leading to inaccurate quantification, poor amplification efficiency, or complete reaction failure.

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IC50, EC50 and Kd: What is the Difference and Why Do They matter?

A modern computer monitor displays a data analytics graph with an upward-trending line in orange and red. The screen has a dark theme with a grid overlay and numerical values. The monitor is set on a desk with a keyboard and mouse, illuminated by warm ambient lighting in the background, creating a professional, high-tech atmosphere.

Three of the most common metrics in drug discover are Kd, IC50 and EC50. At first glance it can seem that they measure the same thing, but they don’t. Kd measures how tightly a molecule or compound binds to its target. IC50 measures inhibition of a function and conversely, EC50 measures activation or induction of a response. Confusing these values can lead to misinterpretation of assay results and costly rework. Let’s take a closer look at each one.

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Glo-ing Above and Beyond: Simplifying Science with MyGlo Reagent Reader

Introduction

When it comes to laboratory tools, few things resonate more than the experiences of researchers who rely on them daily. At the University of Cincinnati the MyGlo Reagent Reader has quickly become an indispensable lab companion, due to its compact design, affordability, and intuitive interface with tailored apps for Promega assays. But what truly sets the MyGlo Reagent Reader apart is how it empowers scientists to focus on their research.

Take Ipsita Kundu, a third-year PhD student at the University of Cincinnati working in Dr. Tim Phoenix’s lab. The Phoenix lab, dedicated to studying innovative brain tumor therapies, faced challenges with their outdated luminescence reader. They needed an affordable, reliable solution to streamline Ipsita’s experiments without compromising accuracy or efficiency.

The MyGlo Reagent Reader was the answer. This blog highlights how this integrated solution is redefining laboratory workflows, enabling researchers to maximize productivity and maintain focus on groundbreaking discoveries. Let’s delve into Ipsita’s story and explore how MyGlo transformed her research.

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Understanding and Combating Legionella in Water Systems with Viability PCR

Water plays a vital role in countless aspects of daily life—drinking, cooling, recreation and more. However, the same systems that deliver these benefits can also harbor Legionella, a waterborne bacteria responsible for Legionnaires’ disease, a severe form of pneumonia (1). Legionella thrives in stagnant aquatic environments, many of which are human-made and common in modern infrastructure, like in cooling towers, hot tubs and complex building water systems. In this blog, we explore the risks posed by Legionella, the limitations of traditional detection methods and how advanced tools at Promega are transforming water safety monitoring. 

3D illustration showing legionella pneumophilia bacteria in water
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The Greatness of Glycogen: A Central Storage Molecule in Energy Metabolism

Introduction

Glycogen is a fundamental molecule in energy metabolism, serving as the critical storage form of glucose that supports cellular health and energy homeostasis. As a polysaccharide, glycogen is essential for maintaining stable energy levels, particularly during periods of fasting and physical exertion. This article will examine glycogen’s synthesis, storage, and utilization, along with its broader significance in human health and disease. Understanding glycogen’s role can provide valuable insight into energy regulation and metabolic health.

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The Benefits of BCAAs: Branched-Chain Amino Acids in Health and Disease

Introduction

Branched-chain amino acids (BCAAs) are essential nutrients that play a significant role in muscle metabolism and overall health. Comprised of leucine, isoleucine, and valine, BCAAs cannot be synthesized by the body and must be obtained through diet. Recent research has highlighted how the metabolic pathways are influenced by BCAAs, such as their ability to activate mTOR signaling, which is vital for muscle protein synthesis (Choi, 2024). Beyond muscle growth, BCAAs may support cognitive function and metabolic health, with ongoing research exploring their broader benefits in disease management. This article explores the diverse roles of BCAAs and their impact on health and diseases

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The Brilliance of BHB: A Key Ketone Body in Metabolic Health

Introduction

β-Hydroxybutyrate (BHB), the most abundant ketone body, is a crucial molecule that sustains energy production during periods of glucose deprivation. Whether you are fasting, adhering to a ketogenic diet, or simply interested in metabolic flexibility, BHB offers key insights into how our bodies adapt to alternative energy sources. This article will delve into how BHB is produced, the diverse roles it plays, and its implications for health and disease.

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Unlocking the Secrets of ADP-Ribosylation with Arg-C Ultra Protease, a Key Enzyme for Studying Ester-Linked Protein Modifications 

Post-translational modifications of proteins are critical for proper protein function. Modifications such as phosphorylation/dephosphorylation can act as switches that activate or inactivate proteins in signaling cascades. The addition of specific sugars to membrane proteins on cells are critical for recognition, interaction with the extracellular matrix and other activities. While we know volumes about some types of protein modifications, ADP-ribosylation on aspartate and glutamate residues has been more difficult to study because of the chemical instability of these ester-linked modifications. 

Matić Lab (Eduardo José Longarini and Ivan Matić) recently published a study that explored mono-ADP-ribosylation (ADPr) on aspartate and glutamate residues by the protein PARP1 and its potential reversal by PARG. PARP1 and PARG signaling are central to DNA repair and apoptosis pathways, making them potentially powerful therapeutic targets in cancer or neurodegenerative diseases in which DNA repair processes are often disrupted. 

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Visualize Protein:Protein Interactions with Bioluminescence Imaging

If you’re familiar with bioluminescence, you’ve probably used it in plate-based experiments to track various biological processes. You understand it provides distinct advantages over traditional fluorescence assays, particularly when it comes to sensitivity. However, there’s always that one nagging question: how representative is the signal on a cell-to-cell level?

Traditional approaches to decipher cell-to-cell signal rely on complex, time-intensive measures that only approximated the findings acquired through bioluminescence. That’s where the GloMax® Galaxy Bioluminescence Imager comes in. This new tool will enhance your ability to visualize proteins using NanoLuc® technology, going beyond simple numeric outputs to reveal what’s happening in individual cells.

NanoLuc® technology is well-known for its ability to assist in detecting subtle protein interactions in complex biological systems. This bright luminescent enzyme enables a much broader linear range than fluorescence, improving detection of small changes in protein activity, such as proteins interacting. Microplate readers measuring NanoLuc® assays rely on signal generated from many cells. This results in an approximation of what is occurring biologically. Truly validating those luminescent readings within a cell population has been challenging—until now. The GloMax® Galaxy allows you to perform bioluminescence imaging, moving beyond the numbers, offering the power to visualize protein interactions directly.

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