Spiroplasma 16S rDNA in TSE Brain Tissues

TSE 脑组织中的螺原体 16S rDNA

• 批准号: 7262853
• 负责人: FRANK O BASTIAN
• 金额: $ 5.01万
• 依托单位: LOUISIANA STATE UNIV AGRICULTURAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7262853
• 关键词: Mycoplasmatales; antibacterial antibody; bacteria infection mechanism; bacterial DNA; bacterial genetics; bacterial proteins; biopsy; brain cell; clinical research; disease /disorder model; genetic screening; genetic strain; human subject; nucleic acid sequence; oligonucleotides; pathologic process; polymerase chain reaction; prions; recombinant DNA; ribosomal DNA; scrapie; spongiform encephalopathy

DESCRIPTION (Adapted from applicant's abstract): The pathogenesis of the transmissible spongiform encephalopathies (TSE), which include Creutzfeldt Jakob disease (CJD) in humans and scrapie in sheep, remains an enigma. In this application we present evidence for the association of Spiroplasma sp., a wall-less prokaryote, with TSE. We have shown PCR amplification of Spiroplasma 16S rDNA in TSE-infected brain tissues (19) and not in control brains (0 of SO). Direct sequencing of the amplified PCR products has confirmed the presence of Spiroplasma-like DNA in all 19 of the TSE brains. Our evidence is not necessarily in conflict with involvement of a PrPres, a protease resistant host derived protein referred to as the prion, in the pathogenesis of TSE, since there is evidence that another factor is involved, which we propose to be a bacterium. Our strategy will be to optimize our probe by characterizing the entire Spiroplasma ribosomal gene in TSE and, then, screen a statistically significant number of TSE cases. In Aim 1, we will utilize a combination of universal Mollicute 16S rDNA oligonucleotide primers along with Spiroplasma-specific 16S rDNA primers to identify by PCR the near complete Spiroplasma 16S RNA gene in TSE. Direct sequencing of the PCR products will ascertain the unknown adjacent portions of the Spiroplasma-like ribosomal gene involved in TSE, from which we will design new oligonucleotide primer set/s that will detect all Spiroplasma strains, including those associated with TSE. In Aim 2 we will screen, with the new probe/s, a sizable collection of TSE cases, including 25 available CJI) human cases and 105 scrapie-infected and normal sheep brains. An additional 100 brain samples, both TSE-infected and normals, are to be sent by collaborators as randomly coded samples. The PCR amplified products in all instances will be sequenced to determine the precise nature of the Spiroplasma sp. involved. Although the role of Spiroplasma in TSE cannot be determined from these experiments, the presence of this microbe in all cases of TSE and not in controls would provide the basis for developing a test for TSE

描述（改编自申请人的摘要）： 可透射的海绵脑病（TSE），其中包括Creutzfeldt 人类中的雅各布病（CJD）和绵羊的crap虫仍然是一个谜。在这个 应用我们提出证据证明螺旋体sp。的关联，a 无壁原核生物，带有TSE。我们已经显示了螺旋体的PCR扩增 TSE感染的脑组织中的16S rDNA（19）而不是对照脑（0 所以）。放大PCR产品的直接测序已证实存在 TSE大脑的所有19个螺旋藻样DNA。我们的证据不是 必然与PRPRES的参与冲突，PRPR是一种抗蛋白酶的宿主 在TSE的发病机理中，派生的蛋白质被称为prion，因为 有证据表明涉及另一个因素，我们认为这是一个 细菌。我们的策略是通过表征 TSE中的整个螺旋马核糖体基因，然后在统计上进行筛选 大量TSE病例。在AIM 1中，我们将使用 通用Mollicute 16S rDNA寡核苷酸引物以及 螺旋体特异性16S rDNA底漆可通过PCR识别 TSE中的Spiroplasma 16S RNA基因。 PCR产品的直接测序将 确定螺旋马样核糖体基因的未知相邻部分 参与TSE，我们将设计新的寡核苷酸底漆。 这将检测所有螺旋菌株，包括与TSE相关的菌株。 在AIM 2中，我们将使用新的探针/S筛选大量TSE集合 案件，包括25个可用的CJI）人类病例和105个受生物的案例， 普通的绵羊大脑。另外100个大脑样本，均受到TSE感染和 正常的，合作者将作为随机编码的样本发送。 PCR 在所有情况下，放大产品都将进行测序以确定精确的 螺旋体Sp的性质。涉及。虽然螺旋体在TSE中的作用 无法从这些实验中确定，该微生物在 所有TSE而非控制案件的情况都将为开发一个 测试TSE

项目成果

Linking chronic wasting disease to scrapie by comparison of Spiroplasma mirum ribosomal DNA sequences.

• DOI: 10.1016/j.yexmp.2004.02.002
• 发表时间: 2004-08
• 期刊: Experimental and molecular pathology
• 影响因子: 3.6
• 作者: F. Bastian;S. Dash;R. Garry

Spiroplasma as a candidate agent for the transmissible spongiform encephalopathies.

螺原体作为传染性海绵状脑病的候选药物。

• DOI: 10.1097/01.jnen.0000183553.01458.87
• 发表时间: 2005
• 期刊: Journal of neuropathology and experimental neurology
• 影响因子: 3.2
• 作者: Bastian,FrankO

Safe method for isolation of prion protein and diagnosis of Creutzfeldt-Jakob disease.

分离朊病毒蛋白和诊断克雅氏病的安全方法。

• DOI: 10.1016/j.jviromet.2005.06.024
• 发表时间: 2005
• 期刊: Journal of virological methods.
• 作者: Bastian,FrankO;McDermott,MichaelE;Perry,AnthonyS;Carver,LarryA;Dash,Srikanta;Garry,RobertF
• 通讯作者: Garry,RobertF