Figure 1 Mycoplasma contamination testing points required by domestic and international regulations

Traditional mycoplasma detection methods mainly include culture methods and indicator cell methods. However, due to the long detection cycles or sensitivity issues of these two methods, they lead to extended production or product release cycles for enterprises. With the development of cell and gene therapy, the industry's requirements for the timeliness and sensitivity of mycoplasma detection have become increasingly stringent. Short shelf lives are insufficient to support such long detection periods, which is why the rapid NAT (Nucleic Acid Testing) method has stood out among various mycoplasma detection methods.

Figure 2 Mycoplasma detection methods listed in domestic and international regulations

Based on NAT (nucleic acid amplification techniques), Yeasen has developed a series of rapid mycoplasma detection kits, including the Mycoplasma Real-time qPCR Detection Kit, the MycAway™ Plus-Color One-Step Mycoplasma Detection Kit, and the GMyc-PCR Mycoplasma Test Kit.

It is a product for the rapid qualitative detection of potential mycoplasma contamination in production raw materials, cell banks, viral seeds, viral or cell harvest fluids, and therapeutic cells. It is primarily used in the research, development, production, and release processes of biopharmaceuticals.

Product Qualifications

Regulatory Compliance: Validated according to the requirements of EP2.6.7, JP G3, and USP 63 pharmacopeias, meeting the standards of international authoritative organizations.

Audit Compliance: The product production complies with the ISO13485 quality management system standards and is supported by comprehensive audit documentation.

Quality Assurance: All enzyme raw materials required for the kit are self-produced and have been industrialized, ensuring stable supply.

Technical Experience Accumulation: There is a solid technical foundation in the TaqMan method, and the kit sensitivity can reach 10 CFU/mL and below.

Focus on Product Performance: The Taq enzyme antibody library, with dual-blocking antibodies, enhances the kit's specificity, sensitivity, and stability.

Product Features

Multiple Detection Types: Optimized TaqMan probes can detect up to 183 mycoplasma species.

Easy to Use: The total time for sample preparation and testing is less than 3 hours, eliminating the need for a 28-day wait associated with culture methods.

High Sensitivity: The detection limit is as low as 10 CFU/mL, making it a viable alternative to direct culture methods.

High Specificity: Primers and probes are designed targeting 16S rRNA, ensuring no cross-reaction with closely related species such as clostridia and lactobacilli.

High Safety: The positive control (PC) in the kit is non-infectious, completely eliminating the risk of contamination.

Strong Interference Resistance: The introduction of an internal control (IC) helps to exclude sample interference and abnormalities in reaction preparation, effectively avoiding false negatives.

Figure 3 Workflow for Mycoplasma Detection Using Mycoplasma Real-time qPCR Detection Kit

It employs a unique isothermal amplification technology, offering higher color discrimination. The change from blue-violet (negative) to sky blue (positive) is more easily discernible by the naked eye. Additionally, it includes anti-contamination components to eliminate the occurrence of false positives.

Figure 4: Operation Procedure for Mycoplasma Detection Using MycAway™ Plus-Color One-Step Mycoplasma Detection Kit

Unlike conventional one-step PCR detection methods, the number of primers is increased from one pair to three pairs, targeting the conserved regions of mycoplasma 16S and 23S rRNA for nested PCR amplification. This not only reduces non-specific products but also significantly enhances the sensitivity of the product, allowing for the detection of as few as a single copy of mycoplasma.

、

Figure 5 Operation Procedure for Mycoplasma Detection Using GMyc-PCR Mycoplasma Test Kit

Mycoplasma Removal

In the process of cell culture, regular testing can effectively prevent large-scale mycoplasma contamination. However, if the cells are already contaminated with mycoplasma, they need to be handled promptly. The best practice is to sterilize the contaminated cells under high pressure and then discard them to avoid contaminating other clean cell lines. If the contaminated cells are particularly valuable and it is necessary to remove the mycoplasma contamination, a combination of antibiotics can be used to achieve this goal. Since mycoplasmas lack a cell wall, traditional antibiotics are generally ineffective against them. Therefore, selecting an appropriate mycoplasma removal reagent is crucial.

Yeasen's MycAway™ Treatment (1000×) is a mixed antibiotic preparation that contains three components with antimicrobial effects on mycoplasma: quinolones, tetracycline derivatives, and macrolide antibiotics. This mycoplasma removal reagent achieves effective mycoplasma clearance by inhibiting DNA synthesis and the production of proteins essential for mycoplasma growth, without harming the cells. It maximizes cell salvage and minimizes losses due to mycoplasma contamination.

Product Features

Low Toxicity: Minimal toxicity to cells, ensuring no interference with subsequent cell experiments such as transfection and viability assays.

High Stability: The reagent can be stored for a long time at -20°C and maintain its high efficacy.

Easy to Use: Simply add the product to the mycoplasma-contaminated culture medium and incubate.

Fast-Acting: 'Immediate Results,' with effects visible as quickly as within 3 days.

Broad Spectrum: Capable of removing the majority of mycoplasma types found in laboratories.

Figure 6. Illustration of Mycoplasma Removal Reagent Cytotoxicity Test

Mycoplasma Prevention

In the process of cell culture, mycoplasma contamination often occurs. However, due to its unique characteristics, it is difficult to detect with the naked eye or microscopic equipment. Therefore, it is particularly important to do a good job in mycoplasma prevention. Effective prevention of mycoplasma contamination can be achieved by adding mycoplasma prevention reagents to the culture medium.

At present, mycoplasma in laboratories mainly originates from: cross-contamination between cells, contamination of the working environment or laboratory equipment, poor aseptic techniques by experimenters, contaminated culture media or reagents, and original tissues or organs with mycoplasma contamination. The mycoplasma prevention product series developed by Yeasen features simplicity, convenience, and high stability, making it very suitable for mycoplasma prevention in routine cell line cultures.

Product Information