DNA Fluorochrome Staining Assay with Indicator Cell Line for Sera Products

M-155 Procedural Summary

This test is designed for the Mycoplasma screening of Serum products. The service consists of the inoculation on an indicator cell line followed by the performance of DNA fluorochrome staining assay. The enrichment component of this method enhances detection of non-cultivable Mycoplasma species.

  • For the indirect method with DNA fluorochrome staining with an indicator cell line, a 35 mL sample aliquot is concentrated by high speed centrifugation.
  • The pellet is resuspended in the cell culture media and inoculated onto a mycoplasma-free indicator cell line (Vero).
  • Cell cultures are incubated at 36°C± 1°C with 5% CO2 and 95% air for a period of 3 to 5 days.
  • Following incubation, the slide culture is fixed and stained with a DNA-binding fluorochrome (Hoechst 33258). The extranuclear fluorescence staining pattern is then interpreted with UV-fluorescent microscopy up to 600x magnification.
  • Appropriate positive (≤ 104 CFU/mL) and negative quality controls are processed with each sample.
Shipping Instructions:

Ship the sample frozen or under usual storage conditions. Bionique Testing Laboratories recommends frozen shipment on dry ice. A 35 mL to 50 mL sample aliquot should be frozen in a sterile 100 mL plastic Nalgene screw cap bottle or 35 mL (maximum 40 mL) of sample should be frozen in a 50 mL sterile polypropylene centrifuge tube.  Sample should be shipped overnight in an insulated container with sufficient dry ice to keep frozen for 48 hours. Place the sample vessel inside a plastic bag with crush proof packaging in an appropriate container. Sample can also be shipped ambient or on cold packs in volumes and vessels as described above.  However, be aware that shipping under ambient conditions may lead to sample compromise due to leakage or exposure to cyclic extremes in temperature.

Please view our general sample submission instructions page for details on how to submit your sample(s) online.



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Total Testing Time: 3-5 days
Sample Requirements: Minimum test volume is 35 mL

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