短喙—侏儒综合征病鸭体内鸭圆环病毒的检测及全基因序列分析

发布时间：2018-02-01 19:56

本文关键词： 鸭圆环病毒短喙-侏儒综合征 PCR检测全基因组序列　出处：《山东农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：自2014年下半年以来,山东、安徽、江苏等地的樱桃谷鸭流行一种以大舌、短喙、生长障碍为主要特征的新发疾病,该病主要引起雏鸭上下喙萎缩变短,舌头外伸、肿胀,感染后期胫骨和翅骨易发生骨折,体重明显低于健康鸭只,导致出栏合格率下降,造成较大的经济损失,经陈少莺等人的分离鉴定,确定该病病原为一种新型小鹅瘟病毒(NGPV)或GPV变异株。在用NGPV阳性样品感染鸭只时,仅有少数鸭只可以复制出与自然病例类似的短喙症状,实验室造病率较低,因此该病的发生绝不仅仅是NGPV单独造成的,还有其他致病因素参与其中,因本病有着与家禽圆环病毒相似的临床症状,如鹦鹉喙羽病病毒常常导致患病的鹦鹉出现鸟喙变形、严重脱毛及体重减轻,鹅的圆环病毒引起机体的矮小综合征,并且Du CV的免疫抑制使其与其他疾病具有较高的混合感染率,因此我们怀疑短喙-侏儒综合征病鸭体内协同感染了鸭圆环病毒。为了解鸭圆环病毒在短喙侏儒综合征发病鸭群中的流行变异情况以及危害程度,明确发病鸭群的的鸭圆环基因型及序列,为疾病的防治提供依据,本实验将完成以下研究内容。1.短喙侏儒综合征发病鸭群中鸭圆环病毒的检测本试验从山东、江苏、安徽等地的24个鸭群的147只病死鸭中采取了心、肝、脾、肺、肾等8个组织器官,采用常规PCR方法对其进行圆环病毒的检测,结果显示,Du CV阳性个体感染率达53.06%(78/147),群体感染率为83.3%(20/24),对79只具有典型大舌短喙特征的鸭子进行了Du CV和NGPV的共感染检测,Du CV与NGPV的混合感染率为53.16%(42/79)。2.鸭圆环病毒的全基因测序及序列分析筛选部分阳性样品进行全基因组扩增共获得4个全基因组序列,其中两株基因组大小为1993nt,另两株为1995nt,与已报道的其他毒株基因组大小一致。基因组内包含3个主要的阅读框,V1/rep、C1/cap和C2/ORF3,所得到4个毒株在上述编码区均没有插入或缺失现象存在。根据全基因序列以及两个主要病毒蛋白核苷酸序列绘制的进化树也一致显示所得的4株序列为基因1型。
[Abstract]:Since the second half of 2014, Cherry Valley Ducks in Shandong, Anhui and Jiangsu provinces have developed a new disease characterized by large tongue, short beak and growth disorder. Tongue extension, swelling, tibia and wing bones are prone to fracture at the later stage of infection, weight is significantly lower than the healthy duck, resulting in a decline in the qualification rate, resulting in greater economic losses, identified by Chen Shaoying and others. When infected with NGPV positive samples, only a few ducks could replicate short-beak symptoms similar to those of natural cases. Laboratory disease rate is low, so the occurrence of the disease is not only caused by NGPV alone, there are other pathogenic factors involved, because this disease has similar clinical symptoms as poultry circovirus. Such as parrot beak feather virus often lead to sick parrot beak deformation, severe hair loss and weight loss, goose circovirus caused the body's dwarf syndrome. And the immunosuppression of du CV makes it have a high mixed infection rate with other diseases. Therefore, we suspect that the duck circovirus was co-infected with the short-beaked dwarf syndrome duck. In order to understand the prevalence of duck circovirus in the short-beaked dwarf syndrome infected ducks and the degree of harm. In order to provide basis for prevention and treatment of the disease, the genotype and sequence of duck ring were identified. Detection of Duck circovirus in Ducks with short beak dwarf syndrome. 147 dead ducks from 24 ducks in Shandong, Jiangsu and Anhui provinces were selected in this experiment. Liver, spleen, lung, kidney and other eight tissues and organs were detected by conventional PCR method. The results showed that the infection rate of DuCV positive individuals was 53.0678 / 147). The coinfection rate of du CV and NGPV was detected in 79 ducks with typical large tongue and short beak. The mixed infection rate of du CV and NGPV was 53.16% and 42 / 79). 2. The whole gene sequencing and sequence analysis of duck circovirus were used to amplify the whole genome of duck circovirus. The genome size of two strains was 1993ntand the other two were 1995nt, which were the same as those of other virulent strains reported. The genome contained three main reading frames V1 / rep. C1 / cap and C2 / ORF3. None of the 4 strains were inserted or deleted in the coding region. The phylogenetic tree drawn from the whole gene sequence and the nucleotide sequences of the two main viral proteins also showed that the 4 strains were gene 1. Type.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.32

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