Identify and quantify threatening microbes

LifeCheck DNA qPCR is innovative genetic testing for rapidly determining problematic microbial populations. It enables the detection and accurate quantification of specific microbes without the need for growth media. Our qPCR technology can target specific microbial functional groups as well as individual species of interest.The reported results provide the total count of specific targeted microbes detected in the sample provided.

Identify and quantify threatening microbes

LifeCheck DNA qPCR Package

Primer SetDescription
Base: Total MicrobesBacteria & Archaea
Target: SRBSulfate Reducing Bacteria
Target: MethanogensTotal Methanogens
Target: Iron Related BacteriaIron Reducing Bacteria and Iron Oxidizing Bacteria
Target: Sulfur CyclingConversion of sulfur related compounds
Target: Sulfur Oxidizing BacteriaOxidation of sulfur-related compounds to sulfate
Target: Specific GeneraSpecific microbe of interest

Understanding your LifeCheck DNA qPCR Package

qPCR Primer SetDescription
Total BacteriaA summary count of all the bacteria in a sample.
Total archaeaA summary count of all the archaea in a sample.
Quality ControlsInternal DNA controls are used to ensure accurate and precise qPCR quantification.
Sulfate reducing bacteria
Sulfate reducing bacteria (SRB) gain energy for growth by reducing sulfate (SO42-) to sulfide (H2S). Sulfide production can cause souring of a system.

SRB can also cause MIC by directly removing electrons from steel surfaces, and indirectly by producing corrosive by-products such as sulfide which react with iron to produce iron sulfide (FeS) deposits.
MethanogensThese anaerobic Archaea produce methane during their metabolism.

Can lead to MIC by removing electrons directly from steel surfaces or indirectly through syntrophic interactions with other microbes.
Iron oxidizing bacteriaIron oxidizing bacteria (IOB) gain energy for growth by oxidizing ferrous iron (Fe2+) to ferric iron (Fe3+), resulting in the formation of ferric iron oxides on the steel surface.

These oxides can be a protective physical barrier preventing any further corrosion, however they also result in the formation of oxygen depleted zones leading to electrochemical alterations of the steel surface, and localized, pitting corrosion.
Iron reducing bacteriaIron reducing bacteria (IRB) gain energy for growth by reducing ferric iron (Fe3+) to ferrous iron (Fe2+).

Fe3+ reduction can remove protective oxide coatings, exposing the surface beneath to further corrosion.
Sulfur CyclingA broad spectrum qPCR target evaluating the genetic capacity for microbial conversion of sulfur related compounds.

Supplementary to the total SRB qPCR primer set, the presence of “sulfur cycling” genes indicates a potential for microbial souring and/or MIC.
Sulfur Oxidizing BacteriaA qPCR primer targeting the oxidation of sulfur-related compounds (sulfide, sulfur, thiosulfate) to sulfate.

Often considered a beneficial group of microbes as they function to remove sulfide (H2S) from a system, and are stimulated (used) in nitrate injection strategies for souring control. Recent research has indicated select SOB species can cause MIC by oxidizing sulfide to sulfuric acid (H2SO4).
Thiosulfate Reducing BacteriaA sulfide producing microbe, increasingly common to oil and gas reservoirs stimulated by hydraulic fracturing methods.

Sulfide production occurs via the use of thiosulfate and not sulfate, making this microbe a non-traditional SRB, undetectable by culture media bottle methods.

Sulfide production poses both a souring and corrosion-based risk.

Understand exactly which threatening microbes are present

Take the analysis of Total Bacteria, Total Archaea, Sulfur Reducing Bacteria, Methanogens, Sulfur Oxidizing Bacteria and Thiosulfate Reducing Bacteria one step further with DNA 16S Sequencing and understand fully the who’s who in the zoo.

Frequently Asked Questions

What is the difference between 16S Sequencing and qPCR?

LifeCheck DNA 16S Sequencing characterizes and provides insight into the microbial community present, and in what relative proportion.

LifeCheck DNA qPCR is innovative genetic testing that detects and accurately quantifies specific microbes without the need for growth media.

Why is it difficult to grow bacteria?

The idea of unculturable microbes first appeared around 100 years ago, when it was observed that cell counts under a microscope were much higher than the number of colonies that would grow on a plate. It has been highlighted again with recent molecular technologies, such as 16S sequencing, which showed us that a single sample can have thousands of different microbes in it.

Think of microbes as tiny people; they have specific conditions required for growth. We cannot survive outside of a narrow temperature range, for example. Microbes also have complex nutrient and environmental requirements. Replicating the conditions required for growth is the biggest challenge, as microbes have diverse pH, nutrient, salinity, temperature. oxygen and pressure requirements.

For example, high temperature microbes, known as thermophiles, will die below a certain temperature, and incubating them at room temperature will kill them. Same with microbes from a high salinity environment. If you put them into culture media with less or different salts than their native environment, they will die. In addition, most microbes live in complex communities where they live syntrophically (they depend on each other) with other microbes, and cannot grow efficiently on their own. Think of it this way, if you were picked up and dropped into the middle of the ocean, how long would you survive? You suddenly don’t have food you can eat, or water you can drink even though you are surrounded by water and fish.

In addition, microbes grow in the natural environment which has a relatively low nutrient concentration, and consequently they are slow growers which can take months or even years to replicate. These microbes may be well established in your system, however trying to grow them to measurable amounts would take a considerable amount of time. In some cases, putting microbes from a nutritionally low to high environment can shock the cells to death. Same reason why a starving person cannot just sit down and eat a large meal after months of not eating, they will become very sick.

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