DEVELOPMENT AND APPLICATION OF THE SYNTHETIC OLIGONUCLEOTIDE PROBE FOR DETECTION OF MYCOPLASMAS IN TISSUE CULTURES.

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It has been concerned that many cell cultures maintained in research,laboratories have been contaminated with various mycoplasmas. This requires a sensitive procedure to detect contaminating mycoplasmas in cell cultures. We have developed a synthetic ologonucleotide probe to detect mycoplasmas by nucleic acid hybridization. The oligonucleotide is a complementary molecule to the region of the ribosomal RNA. The probe consists of 24 bases, 5'-CTAACTTCTGTGTTCGGCATGGGA-3', and is labeled with biotin or alkaline phosphatase. The non-radioactive probes, however, are less sensitive than the radio-labeled one. The biotin labeled probe revealed non-specific reactins. The alkaline phosphatase molecule was enough large to interfere with the hybridization. Thus we are now thinking an alternative procedure such as the polymerase chain reaction (PCR) for this purpose.