[Objective] Torque teno virus （TIT） is a novel virus with negative single-strand DNA discovered in recent years, which is ubiquitous and nonpatho- genie. Torque teno sus virus （TrsuV） is widely prevalent in swine ...[Objective] Torque teno virus （TIT） is a novel virus with negative single-strand DNA discovered in recent years, which is ubiquitous and nonpatho- genie. Torque teno sus virus （TrsuV） is widely prevalent in swine populations, which is considered to be associated with some diseases such as post-weaning multi- systemic wasting syndrome （PMWS）. This study aimed to provided data for epidemiology of Tl＇suV in Guangdong Province. [Method] PCR primers were synthe- sized based on untranslated region （UTR） segment of TYsuV genome, to conduct PCR detection of 14 swine serum samples from two swine farms in Guangdong Province. A total of four PCR products of TrsuV1 and TrsuV2 from two swine farms were selected for cloning, sequencing and analysis. [ Result] Ten TrsuVl pos- itive samples （71% ） and eight TrsuV2 positive samples （57%） were obtained by PCR, including five double-positive samples （36%）. Sequence analysis of PCR products and reference strains showed that the UTR segments of samples GDTI-1 and GDT1-2 were both 305 bp, sharing 90.2% -95.1% similarity with Tl＇suVl reference strain, and the UTR segments of samples GDT2-1 and GDT2-2 were respectively 259 bp and 248 bp, sharing 67.3% - 100% similarity with TrsuV2 refer- ence strain. [ Conclusion] These results conformed that there are at least two types of Tl＇suV in Guangdong Province, with relatively high detection rates in some swine farms. Despite the putative harmlessness of Tl＇suV, the public health significance of TTsuV is noticeable due to its potential pathogenicity.展开更多
为明确福建省猪细环病毒(porcine torque teno sus virus,PTTSuV)1b型(PTTSuV-1b)ORF3基因的遗传进化特征,本研究根据GenBank中登录的PTTSuV-1b基因组特征设计特异性引物,对福建省某猪场患有仔猪断奶后多系统衰竭综合征(PMWS)的猪血清进...为明确福建省猪细环病毒(porcine torque teno sus virus,PTTSuV)1b型(PTTSuV-1b)ORF3基因的遗传进化特征,本研究根据GenBank中登录的PTTSuV-1b基因组特征设计特异性引物,对福建省某猪场患有仔猪断奶后多系统衰竭综合征(PMWS)的猪血清进行PTTSuV-1b分段扩增,并分别对PCR扩增产物进行胶回收后克隆测序,将测序结果经BLAST分析后进行序列拼接。试验结果表明,所扩增的目的片段编码有完整的PTTSuV-1bORF3蛋白,全长为600bp,编码有199个氨基酸。将获得的PTTSuV-1b型福建株与GenBank中PTTSuV-1b型的ORF3基因进行比对分析,其与FJ/China/2010/TTV2/2株核苷酸同源性最高,为99.7%,与西班牙PTTSuV-1b分离株TTV2_G43核苷酸同源性为97.3%,与SC株核苷酸同源性稍低,但也达94.0%;而与猪细环病毒K2型德国家猪分离株472142株核苷酸同源性仅为60.7%,与猪细环病毒1a型西班牙分离株PTTV1_1914株核苷酸同源性仅为46.8%。从遗传进化关系上看,PTTSuV-1bORF3基因在遗传进化上呈2个大的遗传进化分支(分支Ⅰ和分支Ⅱ),本研究分离株处于分支Ⅰ。展开更多
为建立一种特异和快速的猪细环病毒(TTSuV)检测方法,本研究通过比对TTSuV的全基因序列,选择其保守区域,设计了针对TTSuV检测的LAMP引物,利用LAMP Real Time Turbidimeter LA-320仪对反应体系及条件进行了优化,建立TTSuV环介导等温扩增...为建立一种特异和快速的猪细环病毒(TTSuV)检测方法,本研究通过比对TTSuV的全基因序列,选择其保守区域,设计了针对TTSuV检测的LAMP引物,利用LAMP Real Time Turbidimeter LA-320仪对反应体系及条件进行了优化,建立TTSuV环介导等温扩增的检测方法。结果表明:该方法最佳反应条件为64℃恒温50 min,病毒的最低检出限为5.5拷贝/μL,并且与其他相关传染病无交叉反应。临床样品检测结果显示,猪繁殖与呼吸综合征病毒(PRRSV)和2型猪圆环病毒(PCV2)阳性的样品中TTSuV呈高阳性率,分别为92%和87.3%,显著高于非PRRSV和PCV2阳性的猪群。该方法的建立为快速及特异性的检测猪TTSuV提供了有效的方法。展开更多
基金Supported by Funding Project of Guangdong Science and Technology Department(No.2011B010500023)Funding Project of Guangzhou Science and Information Technology Department(No.12A64071507)
文摘[Objective] Torque teno virus （TIT） is a novel virus with negative single-strand DNA discovered in recent years, which is ubiquitous and nonpatho- genie. Torque teno sus virus （TrsuV） is widely prevalent in swine populations, which is considered to be associated with some diseases such as post-weaning multi- systemic wasting syndrome （PMWS）. This study aimed to provided data for epidemiology of Tl＇suV in Guangdong Province. [Method] PCR primers were synthe- sized based on untranslated region （UTR） segment of TYsuV genome, to conduct PCR detection of 14 swine serum samples from two swine farms in Guangdong Province. A total of four PCR products of TrsuV1 and TrsuV2 from two swine farms were selected for cloning, sequencing and analysis. [ Result] Ten TrsuVl pos- itive samples （71% ） and eight TrsuV2 positive samples （57%） were obtained by PCR, including five double-positive samples （36%）. Sequence analysis of PCR products and reference strains showed that the UTR segments of samples GDTI-1 and GDT1-2 were both 305 bp, sharing 90.2% -95.1% similarity with Tl＇suVl reference strain, and the UTR segments of samples GDT2-1 and GDT2-2 were respectively 259 bp and 248 bp, sharing 67.3% - 100% similarity with TrsuV2 refer- ence strain. [ Conclusion] These results conformed that there are at least two types of Tl＇suV in Guangdong Province, with relatively high detection rates in some swine farms. Despite the putative harmlessness of Tl＇suV, the public health significance of TTsuV is noticeable due to its potential pathogenicity.
文摘为明确福建省猪细环病毒(porcine torque teno sus virus,PTTSuV)1b型(PTTSuV-1b)ORF3基因的遗传进化特征,本研究根据GenBank中登录的PTTSuV-1b基因组特征设计特异性引物,对福建省某猪场患有仔猪断奶后多系统衰竭综合征(PMWS)的猪血清进行PTTSuV-1b分段扩增,并分别对PCR扩增产物进行胶回收后克隆测序,将测序结果经BLAST分析后进行序列拼接。试验结果表明,所扩增的目的片段编码有完整的PTTSuV-1bORF3蛋白,全长为600bp,编码有199个氨基酸。将获得的PTTSuV-1b型福建株与GenBank中PTTSuV-1b型的ORF3基因进行比对分析,其与FJ/China/2010/TTV2/2株核苷酸同源性最高,为99.7%,与西班牙PTTSuV-1b分离株TTV2_G43核苷酸同源性为97.3%,与SC株核苷酸同源性稍低,但也达94.0%;而与猪细环病毒K2型德国家猪分离株472142株核苷酸同源性仅为60.7%,与猪细环病毒1a型西班牙分离株PTTV1_1914株核苷酸同源性仅为46.8%。从遗传进化关系上看,PTTSuV-1bORF3基因在遗传进化上呈2个大的遗传进化分支(分支Ⅰ和分支Ⅱ),本研究分离株处于分支Ⅰ。
文摘为建立一种特异和快速的猪细环病毒(TTSuV)检测方法,本研究通过比对TTSuV的全基因序列,选择其保守区域,设计了针对TTSuV检测的LAMP引物,利用LAMP Real Time Turbidimeter LA-320仪对反应体系及条件进行了优化,建立TTSuV环介导等温扩增的检测方法。结果表明:该方法最佳反应条件为64℃恒温50 min,病毒的最低检出限为5.5拷贝/μL,并且与其他相关传染病无交叉反应。临床样品检测结果显示,猪繁殖与呼吸综合征病毒(PRRSV)和2型猪圆环病毒(PCV2)阳性的样品中TTSuV呈高阳性率,分别为92%和87.3%,显著高于非PRRSV和PCV2阳性的猪群。该方法的建立为快速及特异性的检测猪TTSuV提供了有效的方法。
文摘为建立检测猪细环病毒k2型（Porcine Torque teno sus virus k2,PTTSu Vk2）SYBR GreenⅠ实时荧光定量PCR方法,本研究根据PTTSu Vk2 ORF2基因序列特征设计引物,建立基于SYBR GreenⅠ检测PTTSu Vk2的实时荧光定量PCR方法,该方法在PTTSu Vk2 ORF2基因含量为1.92×102~1.92×107 copies/μL反应范围内有很好的线性关系。扩增产物溶解曲线分析仅出现单一特异峰,无引物二聚体,Tm值为（83.83±0.08）℃,对猪圆环病毒（Porcine circovirus type 1 and type 2,PCV1和PCV2）、猪细小病毒（Porcine parvovirus,PPV）、猪博卡病毒5型（Porcine bovavirus type 5,Pbo V5）、猪细环病毒1a型（PTTSu V 1a）和1b型（PTTSu V 1b）核酸均无阳性信号扩增,可重复性好,组内变异系数为0.39%~1.69%,组间变异系数0.69%~2.19%。本方法的建立可用于定量分析PTTSu Vk2感染噬性组织器官和感染程度,为PTTSu Vk2的流行病学研究及临床诊断等提供了一种可靠的方法。
