THE SWIFT DETECTION OF MYCOPLASMA CONTAMINATION IN CELL CULTURES BY USING POLYMYMERASE CHAIN REACTION

Authors

Vet Serum Vaccine Research Institute, Agriculture Research Center, Cairo, Egypt

Abstract

A total of 20 Samples (9 cultures, 7 sera and 4 medium samples) were used in this study. The stained culture samples showed pathological profile resembled that produced by Mycoplasmas. The cell aberrations, clumpings and the increased granularity came out as a result of the Myoplasma impairment influence on the amino acids, RNA and DNA synthesis of the cells which in turn reflected on their metabolic activities. Using the ordinary culturing technique, only 3 cultures and one serum sample gave the characteristic Mycoplasma colonies 4-6 weeks post culturing. The same samples with two cultures and 2 serum samples more were positive with Mycoplasma Polymerase Chain Reaction (PCR), The test was performed and the results were obtained in the same day. The positive bands on Agarose Gel Electrophoresis were of three sizes. A 600bp band seen in rabbit kidney culture samples which might be contaminated with M. orale or M. arginini. The second band (620bp) was observed in VERO, CEF and MDBK cultures that most likely infected with M.hyorhinis or M. fermentans. The third band was 100 bp noticed in the positve serum samples that most probably contaminated with Acholeplasma laidlauii. It could be concluded that PCR technique proved to be accurate, reliable and saving time & can be relied on to detect mycoplasma contamination of cell cultures. Further investigations should be carried out in order to make use of the set in screening of either conventionally produced or imported TC and egg adapted vaccines.

Main Subjects

Veterinary Medical Journal (Giza)
Volume 46, Issue 4
October 1998
Pages 773-783

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