Validation of matrices for Mycoplasma detection (qPCR)

Technical information

Utility

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

The European Pharmacopoeia provides in chapter 2.6.7 that molecular qPCR methods can be used to detect Mycoplasma.

According to the European Pharmacopoeia and GMP guidelines, before carrying out Mycoplasma testing on a medium, reagent, or cellular product, it is necessary to validate that the matrix does not interfere with the testing method, leading to false positive or false negative results. For this reason, matrix validation must be performed before using the Mycoplasma qPCR method.

Method

Following the European Pharmacopoeia, the following validations are carried out:

• Specificity (absence of false positive results)

• Sensitivity (detection capability of 10 colony-forming units CFU or less) of the 6 strains recommended by the European Pharmacopoeia

• Reproducibility (2 replicates are performed)

• Absence of PCR inhibition (internal control)

The procedure follows chapter 2.6.7 of the European Pharmacopoeia.

The validation code at the Banc de Sang i Teixits is V-LC-027.

Diagnostic Algorithm

Not applicable.

Results

The expected results are:

• A NEGATIVE result for Mycoplasma in the negative control (without Mycoplasma)

• A POSITIVE result for Mycoplasma in the positive control (with Mycoplasma)

• A POSITIVE result in the amplification of the internal control

• A POSITIVE result for Mycoplasma in the samples (matrix with added Mycoplasma strain, for each of the 6 strains tested)

This expected result indicates that the matrix does not interfere and that Mycoplasma testing by qPCR can be performed. Any deviation from the expected result is out of specification and must be investigated.

Precautions

Not applicable.

Response Time

• 10 days

Specimen information

Sample: Product, medium 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: 75801

Former BST Code: CEL056

Test Description: Matrix validation for Mycoplasma detection

Synonyms: Mycoplasma validation

Section: Microbiology

BST Fee: Check the updated fees here.

Profiles
Not applicable

References

• Farmacopea Europea 11ena edició