The Serum Test

The recognition that foetal calf serum can introduce mycoplasmas into cell cultures requires the preliminary filtration of calf serum batches to exclude any contaminating organisms followed by a confirmatory test for the absence of mycoplasmas.

Prior filtration, if not completely retaining mycoplasmas, should ensure that few viable units contaminate the final serum lot so any test procedure needs to detect contaminants in relatively large volumes.

We offer a method taken from the findings of M.F. Barile and J. Kern (Proc. Soc. Ep. Biol. Med., 1971, 138, 432-437) who detected Mycoplasma arginini at an incubation temperature of 36°C and Acholeplasma laidlawii after many weeks incubation at 32°C.

THE TEST PROCEDURE

25 ml. volumes of serum are inoculated into each of two lots of liquid medium for incubation at 36°C and 32°C with 0.4 ml. of the serum inoculated directly onto agar medium. The 36°C liquid culture is subcultured two times over two weeks and the 32°C liquid culture is subcultured six times over six weeks. The test is complete 49 days after initiation.

Quality control testing of the calf serum liquid medium, and all solid medium lots used includes M. arginini and A. laidlawii in addition to M. hyorhinis, M. pneumoniae, M. orale and M. synoviae.

TESTING OF OTHER SERA

Where, for example, horse serum, pig serum or other serum sources require testing, we would include a mycoplasma species commonly found in the host animal for test media lot validation (e.g. M. equirhinis with horse serum).

We would be pleased to arrange a quotation for alternative test protocols, either as supplied by the client or designed to meet your needs.

THE FUTURE

Mycoplasma Experience scientists could advise serum manufacturers in the development and validation of new and improved procedures for testing sera for mycoplasma contamination.

The guidelines of Good Laboratory Practice are followed in all aspects of our work and copies of the documents used to record test progress and the quality control of media lots will be included with the final report.

Mycoplasma Experience

Return to Vaccine, Serum and Cell Culture Testing Page