类猪圆环病毒p1分子克隆在猪体的组织分布及对脏器的致病分析(附件)【字数：7513】
1类猪圆环病毒 P1 是新发现的一种病原，能引起仔猪出现类似猪断奶后多系统衰竭综合征症状，是目前所知的具有最小基因组的病毒。为了解P1分子克隆在猪体的组织分布及对脏器的致病作用，构建类猪圆环病毒P1的分子克隆并选用9头仔猪进行人工攻毒，再选取2头阴性对照猪，共11头仔猪，攻毒28天以后采集血清、鼻拭子、心脏、肝脏、脾脏、肺脏、肾脏、胰脏、脑、腹股沟淋巴结等组织。在确定目标病毒的感染后，选取病变组织制作HE病理切片，并且使用荧光定量PCR方法检测病毒在组织中的分布及病毒载量。结果显示，类猪圆环病毒因子P1攻毒成功，病毒主要分布在仔猪的肝脏、肺脏、脾脏、肾脏、胰脏、脑、腹股沟淋巴结等组织。其中病毒载量以肝脏、脑、肺脏较高，且病理切片观察到肝肺肾的异常病变。该研究为P1的诊断及致病机制奠定了基础。
目 录
Abstract1
Key words1
引言2
1材料与方法3
1.1 实验动物与质粒3
1.2 试剂3
1.3 仪器3
1.4 引物3
1.5 实验动物的接种和样本的采集3
1.5.1 实验动物的接种3
1.5.2 不同时期样本的采集3
1.6 样本的病毒分布和病理观察3
1.6.1 病毒DNA的分布检测3
1.6.2 病毒在组织脏器中的定量分析4
1.6.3 病理切片的制作与病理观察4
2结果与分析4
2.1 感染后病毒在组织脏器的分布4
2.2 荧光定量PCR 6
2.3 病理观察7
3讨论8
致谢9
参考文献9
类猪圆环病毒P1分子克隆在猪体的组织分布及对脏器的致病分析
动物医学专业学生 朱雪超
指导教师 宋小凯
Tissue Distribution and Pathogenicity of Molecular Cloning of Porcine CircovirusLike Virus P1 in Infected Pigs
Student ma *好棒文|www.hbsrm.com +Q: ^351916072*
joring in Veterinary Medicine Xue chao Zhu
Tutor Xiao kai Song
Abstract：Porcine circoviruslike virus P1 is a newly discovered pathogen causing symptoms of multiple system exhaustion syndromes in pigs after weaning. It is the smallest genome known to be present. To understand the tissue distribution and pathogenicity of P1 molecular clones, nine piglets were infected with the virus, and two piglets were selected as negative control. Twentyeight days post infection, all the piglets were sacrificed and some tissue samples were collected (including heart, spleen, lung, kidney, pancreas, brain, inguinal lymph nodes, blood and nasal swab). After the infection of the target virus was determined, the diseased tissues were selected and HE staining sections were made. After that, the distribution and viral load of viruses in tissues were detected by fluorescent quantitative PCR. The results showed that all the piglets were confirmed to be infected and the porcine circoviruslike virus P1 was mainly distributed in the liver, lung, spleen, kidney, pancreas, brain and inguinal lymph nodes of the piglets. The viral load was higher in liver, brain and lung, and pathological changes of liver, lung and kidney were observed in pathological section. This study laid the foundation for the diagnosis and pathogenesis of P1.
Key words: porcine circoviruslike virus P1; tissue distribution; pathogenic analysis
猪圆环病毒（procine circocirus，PCV）与鸡贫血病毒（CAV）、鹦鹉喙羽病毒（PBFDV）和一些植物病毒同属于圆环病毒科（Circoviridae）成员，为单股负链环状DNA病毒，直径1517nm，为已知最小的动物病毒之一。现已知PCV有两个血清型，即PCV1和PCV2。
PCV1首次在PK15细胞中被当作是一种细胞污染物而被发现[1]，对猪不具有致病性[2]。而于1997年在法国Clark等研究人员从加拿大西部患有断奶仔猪多系统衰竭综合征（PMWS）的猪群中[3]首次分离得到PCV2型圆环病毒，其主要存在于患僵猪综合征的仔猪，而其所致的疾病称为断奶仔猪多系统衰竭综合征（PMWS）。PCV2的感染还与繁殖障碍、皮炎与肾病综合征、呼吸道疾病等有一定关系，并统称为猪圆环病毒相关疾病，其主要的临床症状主要表现为体质下降，消瘦，贫血，黄疸，生长发育不良，腹泻，呼吸困难等。由于其流行迅速，并且在我国常见多发，因而给养猪业带来了巨大损失[4]。
目 录
Abstract1
Key words1
引言2
1材料与方法3
1.1 实验动物与质粒3
1.2 试剂3
1.3 仪器3
1.4 引物3
1.5 实验动物的接种和样本的采集3
1.5.1 实验动物的接种3
1.5.2 不同时期样本的采集3
1.6 样本的病毒分布和病理观察3
1.6.1 病毒DNA的分布检测3
1.6.2 病毒在组织脏器中的定量分析4
1.6.3 病理切片的制作与病理观察4
2结果与分析4
2.1 感染后病毒在组织脏器的分布4
2.2 荧光定量PCR 6
2.3 病理观察7
3讨论8
致谢9
参考文献9
类猪圆环病毒P1分子克隆在猪体的组织分布及对脏器的致病分析
动物医学专业学生 朱雪超
指导教师 宋小凯
Tissue Distribution and Pathogenicity of Molecular Cloning of Porcine CircovirusLike Virus P1 in Infected Pigs
Student ma *好棒文|www.hbsrm.com +Q: ^351916072*
joring in Veterinary Medicine Xue chao Zhu
Tutor Xiao kai Song
Abstract：Porcine circoviruslike virus P1 is a newly discovered pathogen causing symptoms of multiple system exhaustion syndromes in pigs after weaning. It is the smallest genome known to be present. To understand the tissue distribution and pathogenicity of P1 molecular clones, nine piglets were infected with the virus, and two piglets were selected as negative control. Twentyeight days post infection, all the piglets were sacrificed and some tissue samples were collected (including heart, spleen, lung, kidney, pancreas, brain, inguinal lymph nodes, blood and nasal swab). After the infection of the target virus was determined, the diseased tissues were selected and HE staining sections were made. After that, the distribution and viral load of viruses in tissues were detected by fluorescent quantitative PCR. The results showed that all the piglets were confirmed to be infected and the porcine circoviruslike virus P1 was mainly distributed in the liver, lung, spleen, kidney, pancreas, brain and inguinal lymph nodes of the piglets. The viral load was higher in liver, brain and lung, and pathological changes of liver, lung and kidney were observed in pathological section. This study laid the foundation for the diagnosis and pathogenesis of P1.
Key words: porcine circoviruslike virus P1; tissue distribution; pathogenic analysis
猪圆环病毒（procine circocirus，PCV）与鸡贫血病毒（CAV）、鹦鹉喙羽病毒（PBFDV）和一些植物病毒同属于圆环病毒科（Circoviridae）成员，为单股负链环状DNA病毒，直径1517nm，为已知最小的动物病毒之一。现已知PCV有两个血清型，即PCV1和PCV2。
PCV1首次在PK15细胞中被当作是一种细胞污染物而被发现[1]，对猪不具有致病性[2]。而于1997年在法国Clark等研究人员从加拿大西部患有断奶仔猪多系统衰竭综合征（PMWS）的猪群中[3]首次分离得到PCV2型圆环病毒，其主要存在于患僵猪综合征的仔猪，而其所致的疾病称为断奶仔猪多系统衰竭综合征（PMWS）。PCV2的感染还与繁殖障碍、皮炎与肾病综合征、呼吸道疾病等有一定关系，并统称为猪圆环病毒相关疾病，其主要的临床症状主要表现为体质下降，消瘦，贫血，黄疸，生长发育不良，腹泻，呼吸困难等。由于其流行迅速，并且在我国常见多发，因而给养猪业带来了巨大损失[4]。
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