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

用于检测组织培养中支原体的合成寡核苷酸探针的开发和应用。

• 批准号: 63560290
• 负责人: HARASAWA Ryo
• 金额: $ 1.09万
• 依托单位: UNIVERSITY OF TOKYO (1989)University of Miyazaki (1988)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63560290/
• 关键词: MYCOPLASMA; TISSUE CULTURE; PROBE; プローブ

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.

人们一直关注的是，在研究和实验室中保持的许多细胞培养物已经被各种支原体污染。这需要一种灵敏的程序来检测细胞培养物中受污染的支原体。我们开发了一种人工合成的同源核苷酸探针，通过核酸杂交检测支原体。寡核苷酸是核糖体RNA区域的互补分子。该探针由24个碱基组成，5‘-CTAACTTCTGTGTTCGGCATGGGA-3’，用生物素或碱性磷酸酶标记。然而，非放射性的探测器不如放射性标记的探测器灵敏。生物素标记的探针显示了非特异性反应。碱性磷酸酶分子大到足以干扰杂交。因此，我们现在正在考虑一种替代程序，如聚合酶链式反应(PCR)。

项目成果

Harasawa,R.,Kotani,H.,& Koshimizu,K.: "Genomic heterogeneity of canine ureaplasmass revealed by restriction enadonuclease analysis." journal of Veterinary Medicine,Seris B. 36. (1989)

原泽，R.，小谷，H.，

松崎道哉、原沢亮、君塚房夫、興水馨: Microbiology and Immunology. 33. (1989)

Michiya Matsuzaki、Ryo Harazawa、Fusao Kimizuka、Kaoru Kosui：微生物学和免疫学 33。（1989）。

Matsuzaki,M.,Harasawa,R.,Kimizuka,F.,& Koshimizu,K.: "A non-radioactive DNA prove for the detection of Mycoplasma pulmonis in murine mycoplasmosis." Microbiology and Immunology. 33. 129-132 (1989)

松崎 M.、原泽 R.、君冢 F.、

Matsuzaki,M.,Harasawa,R.,Kimizuka,F.,& Koshimizu,K.: "A non-radioactive DNA probe for the detection of Mycoplasma pulmonis in murine mycoplasmosis" Miccrobiology and Immunology. 33. 129-132 (1989)

松崎 M.、原泽 R.、君冢 F.、

原澤亮(分担執筆): "マイコプラズマとその実験法(輿水・清水・山本編)" 近代出版, 472 (1988)

原泽亮（撰稿人）：《支原体及其实验方法（小清水、清水、山本编）》金代出版社，472（1988）