Mycoplasma Tissue Culture Non-Isotopic Rapid Detection - MTC-NI System

Key Benefits

• Detection and analysis in only 75 minutes including 15 minutes of hands-on time
  
• Detects commonly occurring mycoplasma species with a sensitivity of 10⁵ microorganisms or higher
  
• Detection of multicopy rRNA using Gen-Probe® Hybridization Protection Assay (HPA) technology to ensure sensitivity and robustness
  
• Ideal as a first line screening tool for broad spectrum detection of microorganisms
  
• Easy to use assay requires minimal training

Mycoplasma (e.g microorganisms belonging to class molicutes) contamination is a widespread and reoccurring problem in a wide variety of cell culture systems. These organisms are small (0.2 – 0.3 μm), lack a cell wall and are antibiotic tolerant. This allows them to grow to high titers without exhibiting typical bacterial contamination signs such as a change in turbidity, which traditional growth based methods can not detect. Current methods for positive detection of species belonging to this genus include plating onto agar and liquid co-cultures with VERO cells followed by DNA staining. Although these technologies yield sensitive and reliable results the time to result is typically 2 – 4 weeks.

MTC-NI Technology

The MTC-NI system utilizes the patented HPA assay format from Gen-Probe in which a labeled (acridinium ester) ssDNA probe, complimentary to a conserved region of the ribosomal RNA, is hybridized to the released rRNA of the target organisms. Upon probe binding to the target RNA, the acridinium ester is protected inside the newly formed double helix. After hybridization is completed (all DNA probes have found their target RNA), a selection reagent (0.6M sodium borate) is added to the solution. The selection reagent hydrolyses unbound probes thus circumventing any signal generation from non-hybridized probes. During detection, the bound probes will produce chemiluminescence (induced by hydrogen peroxide from the detection reagent) that is detected by the Leader® instrument. Signal is then presented as Relative Light Units (RLUs) where a positive signal is defined as a value above a certain threshold value.

The main advantages of this technology are the simplicity of the assay both in terms of handling and assay components. Because the MTC-NI HPA assay uses a hybridization event followed by a non-enzymatic hydrolyses and detection, it is very tolerant to sample matrice variation and common inhibitors (Heparin, EDTA etc), which might have a strong negative influence on other common NA detection methods.