1989 Fiscal Year Final Research Report Summary

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

Research Project

Project/Area Number	63560290
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	基礎獣医学
Research Institution	UNIVERSITY OF TOKYO (1989) University of Miyazaki (1988)
Principal Investigator	HARASAWA Ryo UNIVERSITY OF TOKYO, FACULTY OF MEDICINE, ASSOCIATE PROFESSOR, 医学部(医), 助教授 (70159101)
Co-Investigator(Kenkyū-buntansha)	MIZUSAWA Hiroshi NATIONAL INSTITUTE, CELL BANK, OF HYGIENE DIRECTOR, 細胞バンク, 室長
Project Period (FY)	1988 – 1989
Keywords	MYCOPLASMA / TISSUE CULTURE / PROBE
Research Abstract	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.

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