Mycoplasma qPCR

Technical information

Utility

Detection of contaminations by Mycoplasma in samples of advanced therapy, cellular products, cell lines, or culture supernatants, through a Real Time qPCR amplification method, validated by the European Pharmacopoeia.

The determination of contamination by Mycoplasma in starting materials, intermediate products, and final products of cellular medicines is necessary as part of the quality control of the production process, according to Good Manufacturing Practices (GMP).

Method

Mycoplasma is a genus of bacteria that do not have a cell wall. This characteristic makes them resistant to many antibiotics, such as penicillin. Some species of Mycoplasma can cause diseases in humans. Furthermore, these bacteria can contaminate laboratory materials and cell cultures, reducing the yield and viability of the culture, without the contamination being visible under the microscope. According to the European Pharmacopoeia, there are several accepted methods to detect Mycoplasma. At the Blood and Tissue Bank, qPCR (quantitative polymerase chain reaction) is used, a technique that allows the amplification of specific DNA in real time, using fluorescence as a quantification method.

For the extraction of nucleic acids, the QIAamp blood DNA extraction kit (Qiagen) is used. For qPCR, the Venor GeM qEP kit (Minerva Biolabs) is used, with TaqMan probe technology. The technique's sensitivity is ≤10 CFU/mL. The kit includes an internal control that ensures there is no PCR inhibition. Additionally, positive and negative controls are included in each series of tests.

This method has been validated by the Blood and Tissue Bank, with validation code V-LC-030 and validation code for each matrix being V-LC-027.

Diagnostic Algorithm

Not applicable.

Results

The results are reported as POSITIVE or NEGATIVE.

Precautions

Not applicable.

Response Time

• 10 days (can be done in 2 days if urgent, contact the Laboratory)

Specimen information

Sample: Product, media or reagent

Tubes: Sterile tube

Stability: Frozen for 1 month.

Transport instructions: Preferably frozen.

Reason for rejection: Unidentified sample, illegible label, etc.

Administrative information

BST Code: 75800

Old BST Code: CEL055

Test Description: Mycoplasma by qPCR

Synonyms: Mycoplasma

Section: Microbiology

BST Fee: Check the updated fees here.

Profiles
Not applicable

References

• Farmacopea Europea 11ena edició

Quality

BST holds ISO 9001, ISO 14001, and OSHAS 18001 quality certifications, as well as the European Excellence 500+ seal. BST is accredited by CAT, JACIE-FACT, FACT-NETCORD, and EFI, and complies with Good Manufacturing Practice (GMP) and Good Distribution Practice (GDP) guidelines.