1. Introduction: Balancing Speed and Purity in Nucleic Acid Preparation
DNA extraction is a central step in molecular biology, directly impacting the quality and reliability of downstream analyses such as PCR, sequencing, and genotyping. Conventional extraction protocols—often based on silica columns or organic solvents—are known for producing highly purified, double-stranded DNA free of enzymatic inhibitors. However, these methods are relatively time-consuming, labour-intensive, and dependent on multiple reagents and handling steps.
To increase efficiency, rapid lysis methods have been developed that simplify the extraction process by combining chemical lysis and heat treatment in a single-tube format. These protocols typically require less than 20 minutes and only basic laboratory equipment. As a result, they have become widely used in high-throughput or time-sensitive applications, particularly where the extracted DNA is destined for PCR-based detection.
Despite their operational advantages, rapid lysis approaches yield crude lysates that may contain residual proteins, cell debris, or other PCR inhibitors. DNA integrity can also be compromised due to thermal denaturation or fragmentation. In contrast, classical extraction kits, while slower and more resource-intensive, deliver consistently high-purity DNA suitable for a broader range of molecular applications, including sequencing and cloning.
The table below outlines key differences between these two approaches:
| TABLE I – Comparison of Rapid Lysis Methods vs. Classical Extraction Kits | ||
|---|---|---|
| Aspect | Rapid Lysis Methods | Classical Extraction Kits |
| Processing Time | <20 minutes | 30–60 minutes |
| Ease of Use | Simple, single-tube protocol |
Multi-step, hands-on process |
| DNA Purity | Moderate, crude lysate | High purity, inhibitor-free |
| Application Scope | PCR-focused | Broad, including sequencing |
| Cost | Low | Higher |
2. Application Spotlight: Detecting Foodborne Pathogens in Food Samples
Rapid DNA extraction methods have become increasingly important in the detection of foodborne pathogens, including both bacteria and fungi, in complex food matrices. PCR-based techniques are now widely used for routine food safety testing due to their speed and analytical sensitivity. However, their success depends critically on the DNA extraction step, which must yield amplifiable nucleic acids from often challenging and variable sample types.
In food testing workflows, detection typically begins with a pre-enrichment step. Since pathogens in food are often present at low levels, samples are incubated in selective or non-selective broth for 8 to 24 hours to allow viable organisms to multiply. After enrichment, microbial cell counts are significantly increased, generating a high biomass culture suitable for molecular detection.
At this point, the requirements for DNA extraction shift. The primary challenge is no longer sensitivity, but rather speed and efficiency. Rapid lysis methods—based on heat, detergents, or chelating agents—are highly effective under these conditions. They enable nucleic acid release in minutes through a single-tube, boil-and-spin protocol and typically yield DNA of sufficient quality for PCR and qPCR.
The feasibility of using crude lysates is enhanced by the high DNA yield from enriched cultures, as well as the dilution of potential PCR inhibitors in the enrichment medium. As a result, rapid lysis methods are particularly advantageous for high-throughput food safety laboratories, where large sample volumes must be processed under time constraints.
Rapid lysis steps have been successfully integrated into PCR-based detection methods validated according to internationally recognized standards, including AOAC Performance Tested Methods℠ and Microval certifications. These validations—based on rigorous multi-laboratory studies conducted under real-world conditions—evaluate critical performance criteria such as sensitivity, specificity, robustness, and reproducibility. Their inclusion in certified workflows confirms that fast lysis protocols meet the stringent requirements of regulated food testing environments.
These advancements create a strong foundation for the implementation of modern lysis technologies, which combine simplicity and speed with proven regulatory compliance.
3. Our Solution: The InviPrep® Fast Lysis Buffer
The InviPrep® Fast Lysis Buffer is a rapid and efficient reagent for the extraction of PCR-ready DNA from gram-positive and gram-negative bacteria, as well as from fungi. It is validated for use with pure cultures, microbial colonies, and complex matrices such as pre-enriched food and environmental samples. Its simplicity and speed make it highly suitable for both routine screening and regulated laboratory workflows.
The protocol is straightforward and can be completed in about 15 minutes. The buffer is added directly to the sample, followed by incubation at 99 °C for 8 minutes to lyse microbial cells. A short centrifugation step separates debris, yielding a clear supernatant containing DNA ready for use in real-time PCR without further purification (see Figure 1 for workflow overview).
Figure 1: General workflow of DNA extraction using the InviPrep® Fast Lysis Buffer.
Supplied in a ready-to-use format, the buffer requires no additional reagents and contains no guanidinium salts or hazardous solvents. It remains stable at room temperature and delivers consistent performance across a broad range of bacterial and fungal targets.
Its reliability is further demonstrated through integration into validated diagnostic workflows. One example is the InviScreen® Salmonella spp. Detection Kit, which combines the InviPrep® Fast Lysis Buffer with a qPCR-based detection method. This kit is approved under both AOAC Performance Tested Methods℠ (PTM #102401) and Microval (2023LR127) validation programs, confirming that both lysis and detection steps meet international standards for sensitivity, specificity, reproducibility, and robustness.
By combining rapid processing with regulatory-grade performance, the InviPrep® Fast Lysis Buffer offers laboratories a trusted and efficient solution for microbial DNA extraction—supporting accurate pathogen detection in food safety, microbiology, applied molecular diagnostics, and beyond.
4. Uncovering Microbial Warfare: The Dual Strategy of Bacillus subtilis Against Candida albicans
The InviPrep® Fast Lysis Buffer has also proven useful in experimental research settings, supporting investigations into microbial antagonism and biofilm biology. In a recent study by Rajasekharan et al. (2024) [1], the buffer was used in a model of microbial interaction between Bacillus subtilis and Candida albicans to explore how bacterial competition affects fungal growth and virulence.
C. albicans is an opportunistic human fungal pathogen responsible for oral, vaginal, and systemic infections, particularly in immunocompromised individuals. Its ability to form resilient biofilms contributes to antifungal resistance and persistent infections, making it a clinically relevant model organism.
The study revealed that B. subtilis inhibits the growth of C. albicans via a dual mechanism. First, it secretes pulcherriminic acid, which chelates ferric iron and forms pulcherrimin, thereby depleting iron in the local environment and limiting fungal growth. Second, B. subtilis physically colonizes fungal hyphae, interfering with their structural integrity and function. A striking visualization of this antagonism is shown in Figure 2, where biofilm branching toward a C. albicans colony result in pulcherrimin deposition at the interface.
Figure 2: Branching of B. subtilis wild type (WT) colony onto a C. albicans colony (top and bottom views). Blue arrow indicates pulcherrimin build-up in brown color, restricting fungal growth. (Figure from Rajasekharan et al., 2025)
To quantify the effect of these interactions on fungal cell viability, the authors employed a qPCR-based cell viability assay featuring nucleic acid intercalating dyes that selectively enter damaged or dead cells. These dyes covalently bind to DNA, preventing subsequent PCR amplification. DNA was extracted using the InviPrep® Fast Lysis Buffer and then amplified by qPCR, allowing reliable differentiation between live and dead fungal cells. This provided key insights into the suppressive potential of B. subtilis in mixed-species biofilms and further demonstrated the buffer’s utility in advanced microbiological research.
5.Conclusion
Whether used in certified food safety workflows or experimental research, the InviPrep® Fast Lysis Buffer delivers fast, consistent, and PCR-compatible DNA extraction across a wide range of microbial applications. Its robust performance, ease of use, and regulatory validation make it a dependable tool for laboratories seeking to streamline pathogen detection without compromising quality.
References
- Rajasekharan SK, Angelini LL, Kroupitski Y, Mwangi EW, Chai Y, Shemesh M. Mitigating Candida albicans virulence by targeted relay of pulcherriminic acid during antagonistic biofilm formation by Bacillussubtilis. Biofilm. 2024 Dec 18;9:100244. doi: 10.1016/j.bioflm.2024.100244. PMID: 40585313; PMCID: PMC12206338.
