For decades, Oropouche virus (OROV) was considered a problem limited to the Amazon Rainforest. OROV is transmitted to humans and animals through the bite of tiny blood-feeding insects called Culicoides midges. The virus causes Oropouche fever, which leads to debilitating symptoms like a high fever and severe headache1. While the virus was first isolated in Trinidad in 1955, it has since been associated primarily with outbreaks in the Amazon Basin. However, in the last two years, over 29,000 confirmed cases have been reported across the Americas, suggesting the virus has expanded well beyond its historical range.
One of the most concerning expansions is the state of Minas Gerais in southeastern Brazil. Unlike the Amazon, Minas Gerais sits within the Atlantic Forest biome, a heavily fragmented landscape shaped by agriculture, urbanization, and a climate distinct from the humid tropics where OROV has traditionally circulated. With more than 1,600 cases of Oropouche fever since January 2024, a consortium of researchers from universities in Brazil and the US have combined their efforts to try to understand the urgent question: what is driving OROV transmission in this unfamiliar territory2?
RT-qPCR (reverse transcription quantitative PCR) is a powerful technique for quantifying RNA expressionโbut it doesnโt always cooperate. Even when youโve followed the protocol carefully, unexpected results can appear: flat curves, unexpected Cq values, or inconsistent replicates. When that happens, youโre left wondering… what went wrong?
In this blog, weโll walk through five key questions to help you troubleshoot RT-qPCR issues with confidence. From common errors to more stubborn challenges, weโll also explore what to consider when technique isnโt fully the problemโand when it might be time to rethink your reagents.
Legionella is the causative agent of Legionnairesโ disease, a severe form of pneumonia with a mortality rate of around 10%โ. 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 slowโoften requiring 7โ10 days for resultsโand suffer from overgrowth by non-Legionella bacteriaโ. Additionally, traditional methods fail to detect viable but non-culturable (VBNC) bacteriaโcells 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 effortsโ. 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.
Todayโs 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 inhibitorsโsubstances 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.
Our second installment of the Promega qPCR Grant Recipient blog series highlights Dr. Laura Leighton, a trained molecular biologist and postdoctoral researcher at the Australian Institute for Bioengineering and Nanotechnology. Leighton’s scientific journey features a passion for molecular biology and problem-solving. Her path has been illuminated by mentorship, relationships with fellow scientists and a commitment to creativity in overcoming challenges. Here, we explore her scientific journey, reflect on research lessons and foreshadow her plans for the Promega qPCR grant funds.
Dr. Laura Leighton grew up in a rural area in Far North Queensland, Australia, where she spent her early life exploring critters on the family farm. Her upbringing was infused with a deep connection to the environment, from raising tadpoles in wading pools to observing wildlife and witnessing food grow firsthand. Observing the biology around her ultimately piqued her interest in science from a young age. She then began her academic journey in 2011 at the University of Queensland, Australia. She studied biology while participating in a program for future researchers, which led her to undergraduate research work in several research labs. She dabbled in many research avenues in order to narrow in on her scientific interests all while adding different research tools to her repertoire.
After serving as a research assistant in Dr. Timothy Bredyโs lab, she decided to continue work in this lab and pursue a PhD in molecular biology. During her PhD, Leighton worked on several projects from cephalopod mRNA interference to neurological wiring in mice. The common thread in these projects is Leightonโs passion for the puzzles of molecular biology:
โI also love molecular engineering and the modularity of molecular parts. Thereโs something really special about stringing together sequence in a DNA editor, then seeing it come to life in a cell,โ she says.
Dr. Agustรญn Moreira-Saporiti is a postdoctoral researcher at the Marine Biological Laboratory and is studying flowering processes in marine seagrass
Marine seagrasses are submerged flowering plants that form essential underwater meadows, fostering diverse ecosystems and providing a habitat for marine life. Our first Promega qPCR Grant winner and marine ecologist, Dr. Agustรญn Moreira-Saporiti, plans to continue adding to a fascinating body of work aimed at understanding flowering in marine seagrasses.
Dr. Moreira-Saporiti began his journey into marine plant ecology at the University of Vigo, Spain, where he earned a bachelorโs degree in marine sciences. He then went on to complete a masterโs degree at the University of Bremen (Germany) where his thesis focused the ecology of seagrasses in Zanzibar, Tanzania. His passion for marine botany led him down a deeper exploration of marine plants, unraveling the intricate web of ecosystem processes within seagrasses.
Todayโs blog is written by guest blogger, Sameer Moorji, Director, Applied Markets.
People’s diets are frequently influenced by a wide range of variables; with environment, socioeconomic status, religion, and culture being a few of the key influencers. The Muslim community serves as one illustration of how culture and religion can hold influence over people’s eating habits.
Muslims, who adhere to Islamic teachings derived from the Qur’an, frequently base dietary choices on a food’s halal status, whether it is permissible to consume, or haram status, forbidden to consume. With the population of Muslims expected to expand from 1.6 billion in 2010 to 2.2 billion by 2030, the demand for halal products is anticipated to surge (2).
By 2030, the global halal meat market is projected to reach over $300 billion dollars, with Asia-Pacific and the Middle East regions being the largest consumers and producers of halal meat products (3). Furthermore, increasing awareness and popularity of halal meat among non-Muslim consumers, as well as strengthening preference for ethical and high-quality meat, are all contributing to demand.
Foodborne disease affects almost 1 in 10 people around the world annually, and continuously presents a serious public health issue (9).
Food Contamination is common and can be seen in a variety of forms and food products.
More than 200 diseases have evolved from consuming food contaminated by bacteria, viruses, parasites, and chemical substances, resulting in extensive increases in global disease and mortality rates (9). With this, foodborne pathogens cause a major strain on health-care systems; as these diseases induce a variety of different illnesses characterized by a multitude of symptoms including gastrointestinal, neurological, gynecological, and immunological (9,2).
But why is food contamination increasing?
New challenges, in addition to established food contamination hazards, only serve to compound and increase food contamination risks. Food is vulnerable to contamination at any point between farm and tableโduring production, processing, delivery, or preparation. Here are a few possible causes of contamination at each point in the chain (2):
Production: Infected animal biproducts, acquired toxins from predation and consumption of other sick animals, or pollutants of water, soil, and/or air.
Processing: Contaminated water for cleaning or ice. Germs on animals or on the production line.
Delivery: Bacterial growth due to uncontrolled temperatures or unclean mode of transport.
Preparation: Raw food contamination, cross-contamination, unclean work environments, or sick people near food.
Further emerging challenges include, more complex food movement, a consequence of changes in production and supply of imported food and international trade. This generates more contamination opportunities and transports infected products to other countries and consumers. Conjointly, changes in consumer preferences, and emerging bacteria, toxins, and antimicrobial resistance evolve, and are constantly changing the game for food contamination (1,9).
Hence, versatile tests that can identify foodborne illnesses in a rapid, versatile, and reliable way, are top priority.
Boasting a biomass of roughly 400 million tons, Antarctic krill are a key source of food for a wide array of marine life, including sea birds, seals, penguins and whales. As with the rest of the oceanic ecosystem, krill are subject to rapidly shifting climate conditions, prompting scientists to seek a deeper understanding of how they might adapt to a changing environment.
Facing a general lack of genetic information on the species, Professors Cristiano De Pittร and Gabriele Sales from the Department of Biology at the University of Padova in Italy set out to define the krill transcriptome, or sequences of ribonucleic acid (RNA), and in doing so facilitated the discovery of key gene sequences that may play important roles in krill reproduction and survival.
In recent years, there have been concerns about potential impacts to the krill population from ocean warming and commercial fishing operations. Mapping the krill transcriptome may offer scientists crucial insight into the effects of climate change and anthropogenic activity on the dynamics of the Antarctic ecosystem. Doing so is no small feat. Though krill may be miniscule, their genome is 15 times the size of the human genome.
To this end, the research groups of De Pittร and Sales established the database KrillDB, providing a single resource where scientists can access a comprehensive catalogue of krill genes and RNA transcripts. This database represented a powerful bioinformatic tool for examining molecular processes in krill. Funded in part by the Promega 2019 qPCR Grant Program, researchers subsequently rolled out an updated database, KrillDB2, which includes improvements to the quality and breadth of the sequences covered and the information associated. Their corresponding study, published summer 2022 in Scientific Reports, identified a series of genes involved in the krill molting cycle, the reproductive process and sexual maturation, and included never-before reported insights into the expression of microRNA precursors and their effect on krill physiology.
The 2019 Promega qPCR Grant Program offered recipients $10,000 in free PCR reagents and related products, as well as access to Promega technical services and training teams.
Researcher and awardee Alberto Biscontin said of the grantโs impact on their project: โRNA sequencing approaches allow us to determine the level of expression of thousands of genes with a single experiment. The standard in the field is to define transcript expression levels by quantitative RT-PCR to technically validate RNA-seq results. We have been relying on the GoTaqยฎ qPCR solutions by Promega for years.โ He added, โWe have used the GoTaqยฎ 1-Step RT-qPCR System to compare the level of expression of candidate genes with those obtained from RNA-seq analysis. This allowed us to verify at any time the reliability of our bioinformatics pipelines.โ
In the future, researchers plan to maintain the KrillDB2 database with the latest genome and transcriptome sequencing data, to provide the most comprehensive integrative analysis possible. They intend to develop a multi-crustaceans database to support future comparative genomics studies. The KrillDB2 database may also serve as a model to develop other databases for similar species.
Learn more about the GoTaqยฎ 1-Step RT-qPCR System.
In the nine months since the first cases of COVID-19 were noticed in Wuhan, China, the virus has spread around the globe and infected over 22 million people. As with all emerging infectious diseases, we often find ourselves with more questions than answers. However, through the tireless work of researchers, doctors and public health officials worldwide, we have learned a lot about the virus, how it spreads and how to contain it.
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