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Spiroplasma 16S rDNA in TSE Brain Tissues

TSE 脑组织中的螺原体 16S rDNA

基本信息

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
项目类别：
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

项目摘要

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（改编自申请人摘要）：传染性海绵状脑病（TSE），包括克罗伊茨费尔特氏病人类的雅各布病（CJD）和绵羊的痒病仍然是一个谜。在这在本申请中，我们提供了螺旋体属物种，一无壁原核生物，伴有TSE。我们已经显示了螺旋体的PCR扩增 16S rDNA在TSE感染的脑组织中（19），而在对照组脑组织中（0 SO）。扩增的PCR产物的直接测序证实了在所有19个TSE的大脑中发现了类似螺旋体的DNA我们的证据不是这必然与PrPres（一种蛋白酶抗性宿主）的参与相冲突衍生蛋白称为朊病毒，在TSE的发病机制中，有证据表明，另一个因素是涉及，我们建议是一个细菌。我们的策略是通过表征整个螺旋体核糖体基因在TSE，然后，筛选一个统计学大量TSE病例。在目标1中，我们将结合使用通用柔膜菌16S rDNA寡核苷酸引物沿着螺旋体特异性16S rDNA引物，通过PCR鉴定接近完整的 TSE中的螺原体16S RNA基因。PCR产物的直接测序将确定螺旋体样核糖体基因的未知相邻部分我们将从中设计新的寡核苷酸引物可以检测所有螺原体菌株，包括与TSE相关的菌株。在目标2中，我们将使用新探头筛选大量TSE 病例，包括25例可用的CJI）人间病例和105例羊瘙痒症感染，正常的羊脑另外100个大脑样本，既有TSE感染的，法线，将由合作者作为随机编码的样本发送。的PCR 所有情况下的扩增产物都将被测序以确定精确的所涉及的螺原体的性质。尽管螺原体在TSE中的作用不能从这些实验中确定，这种微生物的存在，所有TSE病例和非对照病例将为制定 TSE检测

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
通讯作者：
F. Bastian;S. Dash;R. Garry

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.
影响因子：
0
作者：
Bastian,FrankO;McDermott,MichaelE;Perry,AnthonyS;Carver,LarryA;Dash,Srikanta;Garry,RobertF
通讯作者：
Garry,RobertF

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。