Validation of a PCR method for the detection of mycoplasmas according to European Pharmacopoeia section 2.6.7.
Recent publication of a revised section for mycoplasma testing in the European Pharmacopoeia has led to interest in validating a nucleic acid amplification technique (NAT) for use in detection of mycoplasma contaminants in biologics drugs. The replacement by or supplementation of the existing culture-based methods with a PCR-based method has several advantages for the biopharmaceutical industry, mainly with respect to reduced turnaround time for results, and potentially lowered cost. Replacement or substitution of existing methods by a PCR method requires the demonstration of equivalent assay limit of detection (LOD) and specificity. The experimental requirements for this comparability validation have been enumerated in detail in the EP section referenced above. In this publication, we describe the validation and comparability analysis of a PCR method performed exactly according to the EP guidance. Completion of this validation activity has resulted in the availability of an assay that meets or exceeds EP compliance requirements for a mycoplasma detection method.
Department of Molecular Biology, WuXiAppTec, Inc., 4751 League Island Boulevard, Philadelphia, PA 19112, USA.
Yan
Y
Zhi
Amy
A
Mayhew
Natalie
N
Seng
Garry B
GB
Takle
D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods / Q000592:standards; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D005060:Europe; D006801:Humans; D057230:Limit of Detection; D009174:Mycoplasma / Q000235:genetics* / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology; D010602:Pharmacopoeias as Topic / Q000592:standards; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013045:Species Specificity
D004269:DNA, Bacterial