2: An Evaluation of Comparability Between an Alternative NAT Method Using Multiplex PCR and the Compendial USP <63> Culture-Based Method

The need to reduce time to release for short shelf-life products has driven the steady advancement of rapid alternative methods in biotechnology. The BIOFIRE® Mycoplasma Panel is an automated molecular diagnostics method that utilizes multiplex PCR and promotes a run time of less than 1 hour for the detection of USP, Ph. Eur. and JP Mycoplasma species.
To verify these claims, Bionique Testing Laboratories (“Bionique”) performed a comparability study between the BIOFIRE® assay and the compendial USP < 63> culture-based method. Recovery of viable mycoplasma from CHO cell suspensions was tested using the 10mL BIOFIRE® Mycoplasma Panel protocol and compared to results obtained using Bionique’s Harmonized USP < 63> test method for the same material. Spike recovery from CHO cell suspensions staged at 5×106 cells/mL was evaluated for ten different strains of mycoplasma at 10 CFU/mL, while simultaneously evaluating non-spiked material for interference. Negative recovery was observed from all non-spiked suspensions and positive recovery from all spiked suspensions across both methods, establishing comparability between these two tests for this particular product. Importantly, the BIOFIRE® System also generated results for a single sample, including prep time, in just over 2 hours compared to the 28 days required for the USP < 63> compendial method.