Diagnosis of streptococcal infection by PCR amplification

PCR 扩增诊断链球菌感染

• 批准号: 06671815
• 负责人: ONO Tsuneko
• 金额: $ 1.15万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671815/
• 关键词: oral streptococci; PCR diagnosis; spaP; S.mutans; bacteremia; Streptococcus.mutans; PCR; spaP gene; 敗血症

We have researched the mechanism of pathogenicity of oral streptococci which caused infectious endocarditis, bacteremia and pneumonia and have developed rapid diagnosis methods of oral streptococcus infection. Streptococcus mutans, a member of viridans streptococci, is recognized as a part of the normal oral flora of man, and is an etiological agent in smooth-surface dental caries. We have tried to detect S.mutans in blood and other clinical specimens at first.Synthetic oligonucleotide primers were used in the polymerase chain reaction to amplify a sequence of the spaP gene, which encodes the surface protein antigen I/II of Streptococcus mutans. A DNA fragment was amplified from lysed S.mutans cells or isolated DNA.With S.mutans cells, the lower limit of detection was 4-40 cfu. With these primers, 13 reference and 50 clinical strains of S.mutans were identified. Amplification of the 200-bp product was not demonstrated when 41 strains of other streptococcal and non-streptococcal species were tested. The spaP gene PCR has potential for the rapid diagnosis of S.mutans infections.

我们对引起感染性心内膜炎、菌血症和肺炎的口腔链球菌的致病机理进行了研究，并建立了口腔链球菌感染的快速诊断方法。变形链球菌是草绿色链球菌的一种，是人类口腔正常植物群的一部分，是光滑面龋齿的病原体。我们首先尝试从血液和其他临床标本中检测变形链球菌，用人工合成的寡核苷酸引物，在聚合酶链反应中扩增编码变形链球菌表面蛋白抗原I/II的spaP基因序列。从变形链球菌裂解细胞或分离的DNA中扩增出一段DNA片段，对变形链球菌细胞的检测下限为4-40 cfu。用这些引物对13株参考菌株和50株临床菌株进行了鉴定。扩增的200-bp的产品没有被证明时，其他链球菌和非链球菌物种的41株进行了测试。spaP基因PCR技术可用于变形链球菌感染的快速诊断。

项目成果

Shigeru Akimoto,et al.: "Complete sequence of the Bacteroides fragilis YCH46 neuraminidase-encoding gene." Biodhem.Biophys.Res.Comm.203. 914-921 (1994)

Shigeru Akimoto 等人：“脆弱拟杆菌 YCH46 神经氨酸酶编码基因的完整序列。”

Mahfuzur R.Sarker, et al: "Molecular Cloning of the leuB gene from Bacteroides fragilis by functional complementation in Escherichia coli." Microbiol.Immunol.39. 19-25 (1995)

Mahfuzur R.Sarker 等人：“通过大肠杆菌中的功能互补对脆弱拟杆菌的 leuB 基因进行分子克隆。”

Katsuhiko Hirota, et al.: "Cross-reactivity between human sialyl Lewis^X oligosaccharide and common causative oral bacteria of infective endocarditis." FEMS Immunol.Med.Microbiol.12. 159-164 (1995)

Katsuhiko Hirota 等人：“人唾液酸 Lewis^X 寡糖与感染性心内膜炎常见口腔细菌之间的交叉反应性。”

Tsuneko Ono, et al.: "Detection of Streptococcus mutans by PCR amplification of spaP gene." J.Med.Microbiol.41. 231-235 (1994)

Tsuneko Ono 等人：“通过 PCR 扩增 spaP 基因检测变形链球菌”。

Shigeru Akimoto, et al.: "Complete sequence of the Bacteroides fragilis YCH46 neuraminidase-encoding gene." Biochem. Biophys. Res. Comm.203. 914-921 (1994)

Shigeru Akimoto 等人：“脆弱拟杆菌 YCH46 神经氨酸酶编码基因的完整序列。”