Safety assays for quality control of ATMPs: addressing mycoplasma detection challenges
Cell & Gene Therapy Insights 2026; 12(3), 273–283
DOI: 10.18609/cgti.2026.035
Published: 29 April
Innovator Insight
In biotherapeutic manufacturing, mycoplasma testing plays an important role in ensuring product safety, but current methods do not always align with the speed required in modern processes. Culture-based assays, although accepted, can take several weeks to generate results. This article looks at how reverse transcription digital PCR (RT-dPCR) can be used as an alternative. The approach enables detection down to 5-10 CFU/mL and performs reliably even in complex sample matrices. When combined with automated nucleic acid extraction, it allows testing to be completed within a single working day.
Culture-based mycoplasma testing takes up to 28 days — incompatible with modern ATMP timelines. This article evaluates RT-dPCR as a same-day, pharmacopeia-compliant alternative.
What you will learn
01
Why RT-dPCR offers superior sensitivity over dPCR and qPCR at low contamination levels
02
How the QIAcuity Mycoplasma Quant Kit meets pharmacopeial thresholds across multiple species
03
How automated extraction enables a complete sample-to-result workflow in a single working day
The RT-dPCR detection workflow
1
Sample preparation
Automated extraction on EZ2 Connect
2
Reaction setup
Eluate added to QIAcuity Master Mix & nanoplate
3
RT-dPCR analysis
Results within 6–7 hours total
Key findings
Detection down to 5 CFU/mL — exceeding the 10 CFU/mL pharmacopeial threshold for replacement of culture-based methods
Reliable performance in complex matrices, with linear quantification confirmed at R² > 0.995
Same-day results when combined with automated EZ2 Connect extraction, supporting faster QC decisions in ATMP manufacturing
Key interests
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