Mycoplasma Detection

PCR, enzymatic-luminescence, and culture methods for mycoplasma detection in cell culture

Mycoplasma is the silent contaminant in cell culture; an estimated 15 to 35 percent of working lines worldwide are infected. Routine testing catches it. This section covers PCR-based detection (most sensitive), enzymatic-luminescence (faster but less sensitive), and direct culture (slow but definitive).

Pick method by sensitivity needs.

1. Highest sensitivity. PCR-based detects 10 to 50 organisms per mL. Catches contamination earliest.
2. Fast surveillance. Enzymatic-luminescence (MycoAlert-equivalent) detects ~10^4 organisms per mL. 20 minutes per sample.
3. Definitive identification. Direct culture. Slow but conclusive.
4. Throughput. PCR for 96-well surveillance. Enzymatic for individual line checks.
5. For high-throughput facility surveillance enzymatic-luminescence catches gross contamination quickly; confirm any positives by PCR.

ABM and Norgen Biotek lead in PCR-based detection kits.