Mycoplasma Testing

Microtest provides experienced, reliable testing for the detection of Mycoplasma contamination in Biological products. The purpose of the test is to determine the presence or absence of mycoplasmal contaminants in the cell cultures samples.

Microtest follows FDA Points-to-Consider (PTC) document for performing these assays. The FDA requires that both assays are performed in order to ensure a high degree of certainty in confirming the presence or absence of Mycoplasma contamination.

The testing challenges consist of the following two methods:

1. Direct cultivation method: This method utilizes mycoplasma broth and agar formulations. A 10 ml product is inoculated into the mycoplasma media. The media is incubated and transferred to agar media plates in duplicate on day 3, 7, and 14. At each transfer, the plates are examined microscopically at day 7 and 14. Control plates utilizing Mycoplasma orale and Mycoplasma pneumoniae are prepared along side the test products.



2. Cell cultured method: One (1) ml of product is inoculated into prepared VERO cell plates in duplicate and incubated for 3-5 days. After incubation, the plates are stained with a DNA Fluorochrome stain. The plates are then examined to determine if Mycoplasma contamination is evident. Control plates of Mycoplasma orale and Mycoplasma hyorhinis are prepared along side the test products.

The test passes if Mycoplasma is not detected using the direct and cell culture methods. The controls must exhibit growth when inoculated with < 100 CFU per organism. All negative controls must be free of Mycoplasma contamination.