Lab Insights: Fewer Steps, Better Lysis – How RTP® Technology Strengthens Pathogen Detection
Part of the Lab Insights series: practical perspectives on molecular workflows from the bench outward.
1. The Lysis Step: Foundation of Every Reliable Result
Ask most laboratory professionals which part of a nucleic acid extraction protocol causes the most problems, and the answer is rarely the wash steps or the elution. It is lysis, and more specifically, the challenge of achieving it consistently across the full spectrum of organisms a diagnostic laboratory is likely to encounter.
Lysis is the point at which sample biology becomes chemistry. Cell walls are disrupted, intracellular contents are released, and the nucleic acids that downstream assays depend on become accessible. If this step is incomplete or poorly matched to the pathogen in question, no amount of downstream optimisation will recover what was lost. A PCR assay with excellent analytical sensitivity will still underperform on poorly extracted material.
The difficulty is that lysis is not one thing. What disrupts a lipid-enveloped virus efficiently will not work equally well on a Gram-positive bacterium with a thick peptidoglycan wall, a fungal spore with its chitinous structure, or a protozoan parasite encased in layers of protective membrane. Each organism carries different structural resistance and demands a different biochemical approach.
In practice, laboratories rarely have the luxury of tailoring their workflow to each individual pathogen. Samples arrive, protocols need to be consistent, and results are expected to be reliable. The question is how to build a lysis approach broad enough to handle the full diversity of pathogens passing through a laboratory, without requiring a different tube for every organism type.
That is the problem RTP® technology was designed to solve.
2. All the Chemistry You Need for Lysis, Pre-loaded in One Tube
RTP® stands for Ready-To-Prep. The core of the technology is a pre-filled Extraction Tube in which all the components required for sample lysis have been lyophilised together (see Fig. 1). When a sample is added to the tube, the lysis chemistry is already there, in the correct quantities and formulation, ready to work. No manual pipetting of individual reagents. No preparation before lysis begins.
Figure 1: RTP® Technology: Extraction Tube with lyophilized lysis components.
Each component addresses a specific challenge in the lysis of structurally diverse pathogens. The Lysis Buffer creates the right chemical conditions; Proteinase K dismantles protein barriers; Lysozyme targets the tough cell walls of Gram-positive bacteria; and Carrier RNA ensures that what has been released is not then lost to surface adsorption before purification begins. The table below summarises the key components and the role each plays in achieving broad-spectrum lysis.
Table 1: Lyophilised components of the Extraction Tubes and their role during lysis.
|
Component |
Function |
|
Lysis Buffer |
Non-chaotropic disruption of membranes |
|
Proteinase K |
Digestion of nucleic acid-binding proteins |
|
Lysozyme |
Breakdown of bacterial cell walls |
|
Carrier RNA |
Stabilization of low-abundance nucleic acids |
The result is a lysis step capable of handling bacteria, viruses, yeasts, moulds, and protozoa from a single, standardised tube format, without requiring separate reagent preparation or organism-specific protocol adjustments.
3. Room Temperature Stability. Less Pipetting. Less Variability.
Lyophilisation is what makes the RTP® format practical beyond the bench. Freeze-drying the lysis components removes water and produces a stable, dry matrix that resists degradation over time and across temperature fluctuations. The Extraction Tubes can be stored at room temperature with no cold chain requirement, no thawing before use, and no freeze-thaw variability to account for before a single sample has been processed. The lyophilised format retains its performance characteristics throughout its shelf life, meaning a tube used on day one of a batch behaves the same as one used several months later.
The same design logic that removes cold chain dependency also removes variability at the lysis step itself. Because the lysis components are pre-loaded in the Extraction Tube, lysis requires no reagent preparation. The operator adds the sample, and the chemistry begins. Compared with conventional manual extraction protocols, this consolidation reduces pipetting steps by up to 60% at the lysis stage (see Fig. 2). Fewer steps mean less hands-on time, lower consumable use, and a reduced opportunity for error. In routine diagnostic settings, that translates into measurable time savings, a lower cost per sample, and a simpler protocol that is easier to train on and easier to validate.
Figure 2: Processing of samples with the RTP® method significantly reduces hands-on-time and pipetting steps compared to the classical method.
One of the defining design principles behind RTP® technology is that the same tube format should work across a wide range of pathogens and sample types without requiring protocol modifications for each combination.
In practice, this means the Extraction Tube handles simultaneous isolation of bacterial DNA, viral DNA, and viral RNA from a single sample. The lysis conditions are optimised to accommodate organisms with fundamentally different structural properties: from fragile enveloped viruses to resilient Gram-positive bacteria, from yeast and mould with their thick cell walls to protozoan parasites with complex membrane architectures.
Sample type coverage is equally broad. The technology is suited to human clinical samples including serum, plasma, swabs, and respiratory material, as well as to veterinary, food, plant, and environmental matrices including wastewater. This range makes the RTP® technology relevant across a wide spectrum of laboratory settings, from hospital microbiology to food safety testing, animal health diagnostics, and environmental surveillance.
For laboratories managing multiple testing programmes or working across application areas, the ability to apply a single extraction format consistently across different sample types reduces the number of protocols in use, simplifies SOP management, and lowers the risk of method-selection errors.
Published data provides a useful measure of how RTP® technology performs under real diagnostic conditions. A study by Kann et al. (Microorganisms, MDPI, 2022) compared three commercial extraction kits using 62 serum reference samples from patients with confirmed Chagas disease, testing all eluates with the same Trypanosoma cruzi-specific real-time PCR assay to isolate extraction as the key variable. T. cruzi is a protozoan parasite, and its DNA concentrations in human serum can be very low, making it a particularly demanding test of extraction performance.
The RTP® Pathogen Kit was the only kit to achieve 100% sensitivity, correctly identifying all 62 positive samples. The comparator kits returned 98.4% (61/62) and 96.8% (60/62) respectively, with missed samples remaining negative on repeat testing, confirming the failures were attributable to extraction. The RTP® Pathogen Kit also produced the lowest mean Ct value, indicating consistently higher target DNA yield from the same sample volume.
Table 2: Results of the study by Kann et al. (Microorganisms, MDPI, 2022). The RTP® Pathogen Kit outperformed other kits in extraction of Trypanosoma cruzi DNA from patients with confirmed Chagas disease.
|
Extraction Kit |
Samples Detected |
Mean Ct Value |
|
RTP® Pathogen Kit (Invitek Diagnostics) |
62/62 (100%) |
30.8 |
|
MagaBio plus Virus DNA/RNA Kit (Bioer Technology) |
61/62 (98.4%) |
33.4 |
|
EZ1&2 Virus Mini Kit v2.0 (Qiagen) |
60/62 (96.8%) |
31.8 |
The technologies capacity for simultaneous extraction of bacterial DNA and viral RNA from a single sample is further illustrated by a study by Krumkamp et al. (Scientific Reports, 2023), which used the RTP® Pathogen Kit to screen pharyngeal swabs from children hospitalised with acute lower respiratory tract infections against a panel of 19 targets, covering bacterial pathogens including S. pneumoniae, H. influenzae, and Chlamydiae, alongside respiratory viruses including influenza A/B, RSV, adenovirus, and rhinovirus. Up to six pathogens were detected concurrently in a single individual, demonstrating the extraction kit's ability to support broad-spectrum detection across structurally diverse organisms from one sample.
Together, these studies reflect a consistent principle: when lysis is thorough and reliable, the downstream assay has the best possible material to work with.
Reliable pathogen detection begins with reliable lysis. RTP® technology addresses this at the source, combining all components required for broad-spectrum lysis in a single, pre-loaded, lyophilised extraction tube. The result is a standardised, room-temperature-stable format that significantly reduces hands-on time and pipetting steps, eliminates cold chain dependency, and delivers consistent extraction performance across a wide range of pathogens and sample types.
For laboratories where accuracy, reproducibility, and workflow efficiency are required, RTP® technology offers a lysis platform designed to meet those demands without compromise.
What Invitek Offers
RTP® technology is available in two extraction kits, each designed for a distinct application domain. Both kits are built on the same lyophilised, pre-filled Extraction Tube format, delivering the same workflow simplification, room temperature stability, and broad-spectrum lysis capability across two distinct regulatory and application contexts:
The RTP® Pathogen Kit is designed for clinical diagnostics and is CE-IVD certified in compliance with EU Regulation 2017/746 on in vitro diagnostic medical devices. It is validated for the isolation of bacterial DNA and viral DNA and RNA from clinical sample types including serum, plasma, urine, swabs, respiratory samples, and tissue. It is the appropriate choice for laboratories operating within a regulated diagnostic framework requiring documented conformity with IVD requirements.
The InviSorb® Lyo Pathogen Kit extends the RTP® technology into non-clinical applications. It supports DNA and RNA isolation from bacteria, viruses, yeasts, moulds, and protozoa across a broad range of matrices including veterinary specimens, food and beverage samples, environmental and wastewater samples, and plant material. It is suited to laboratories working in food safety, veterinary diagnostics, environmental monitoring, and plant pathology, where sample complexity and matrix diversity place particular demands on lysis performance.
[1] Kann S, Zabala-Monterroza W, García C, et al. Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCR. Microorganisms. 2022 Jul;10(8):1554. DOI: 10.3390/microorganisms10081554. PMID: 36013972; PMCID: PMC9414588.
[2] Krumkamp R, Kohsar M, Nolte K, et al. Pathogens associated with hospitalization due to acute lower respiratory tract infections in children in rural Ghana: a case-control study. Scientific Reports. 2023 Feb;13(1):2443. DOI: 10.1038/s41598-023-29410-5. PMID: 36765075; PMCID: PMC9916495.