小鵝瘟病毒PCR檢測方法的建立和山東濟寧地區(qū)小鵝瘟流行病學調(diào)查
發(fā)布時間：2018-10-21 11:52
【摘要】：小鵝瘟是由鵝細小病毒引起的一種急性傳染病,主要侵害30日齡以內(nèi)的雛鵝和雛番鴨,以發(fā)病急,死亡率高,傳播快為特點。發(fā)病率和死亡率隨著日齡的增加而下降,主要侵害7日齡以內(nèi)的雛鵝和雛番鴨,根據(jù)發(fā)病情況分為3種類型,最急性型,急性型和亞急性型(殷震and劉景華,1997)。發(fā)病鵝主要以腸粘膜脫落,壞死,腸栓,脫水等為癥狀。1966年,匈牙利學者Derszy用鵝胚也分離到病毒,他是國外第一個用鵝胚分離到病毒的人,并于1967年對本病進行了詳盡的描述,但當時他卻把該病誤認為流感,直到1971年Schettler確定這種病是由細小病毒引起。1978年建議把這種病稱為鵝細小病毒感染,1974年世界禽類學會為紀念Derzsy的先期工作而將此病命名為Derzsy氏病(Derzsy D,1975)。1995年Zadori等首次報道了GPV全基因序列,并將其與番鴨細小病毒基因序列進行了同源性比較(Zadori Z,1995)。本病大流行具有一定周期性。在大流行以后當年余下的鵝群都獲得了主動免疫,使次年的雛鵝具有天然被動免疫力,故本病不會在同一地區(qū)連續(xù)兩年發(fā)生大流行。由此可見,本病的發(fā)病率和死亡率一方面和日齡有關(guān),另一方面取決于親代母鵝的特異性抗體水平。免疫預防是控制小鵝瘟的重要措施。目前已建立了PCR、ELISA、間接免疫熒光技術(shù)、微量中和試驗等檢測小鵝瘟的方法,但是這幾種檢測方法均需特殊的儀器設備,且耗時長,不適于在基層推廣應用。因此,建立一種更加快速、靈敏的病毒檢測方法迫在眉睫。本研究建立的聚合酶鏈式反應,具有操作簡單,快速,適合基層推廣的特點。近年來小鵝瘟在我國的一些省、市、自治區(qū)均有流行,給養(yǎng)漖業(yè)造成嚴重的危害,同時也給飼主帶來巨大的經(jīng)濟損失,因此受到國內(nèi)外學者和工作者的廣泛關(guān)注。本研究通過工作之便,對所在地區(qū)鵝養(yǎng)殖現(xiàn)狀和小鵝瘟發(fā)病情況做了相應的調(diào)查,并對其流行情況做了匯總,以期對養(yǎng)鵝業(yè)小鵝瘟疾病的發(fā)生做出相關(guān)的預防措施。本研究根據(jù)GENBANK上發(fā)表的小鵝瘟的vp3的序列信息,設計了一對特異性引物,通過優(yōu)化各種條件,最終擴增出與預期片段大小一致的314bp的核酸片段,在通過相應的特異性實驗發(fā)現(xiàn)此引物僅能擴增出小鵝瘟的特異片段,能夠用來檢測小鵝瘟的發(fā)生。本研究中對于分離到的10株病毒進行測序分析,結(jié)果顯示分離株之間,同源性最高達100%,最低也在95.2%以上,說明小鵝瘟病毒的擴增片段核苷酸變異很小,VP3是小鵝瘟的主要抗原,所以本研究對于免疫預防具有指導性的意義。依據(jù)本研究中調(diào)查的小鵝瘟的發(fā)病日齡等基本信息,尤其每個鵝場的免疫預防程序進行了調(diào)查,發(fā)現(xiàn)小鵝瘟的發(fā)病日齡集中在12-2月份之間,根據(jù)小鵝瘟的發(fā)病情況,對其免疫程序作出了相應的調(diào)整,結(jié)果顯示小鵝瘟的發(fā)病概率降低了,說明合適的免疫程序是預防該病的比較有效的措施。
[Abstract]:Gosling plague is an acute infectious disease caused by goose parvovirus, which mainly infects goslings and young muscovy ducks within 30 days of age. It is characterized by rapid onset, high mortality and rapid transmission. The morbidity and mortality decreased with the increase of the age of the day, mainly affecting the goslings and young muscovy ducks within 7 days of age, which were divided into three types according to the incidence, the most acute type, the acute type and the subacute type (Yin Zhen and Liu Jinghua, 1997). In 1966, Hungarian scholar Derszy also isolated the virus from goose embryos. He was the first person in foreign countries to isolate the virus from goose embryos, and described the disease in detail in 1967. But at the time he mistook the disease for the flu, It was not until 1971 that Schettler determined that the disease was caused by parvovirus. In 1978, it was suggested that the disease should be called goose parvovirus infection. In 1974, the World Avian Society named the disease Derzsy's disease (Derzsy Dl1975) to commemorate the advance work of Derzsy. Zadori et al. The entire GPV gene sequence, The gene sequence of muscovy duck parvovirus was compared with that of muscovy duck parvovirus (Zadori ZJ 1995). The epidemic has a certain periodicity. After the pandemic, the remaining geese got active immunity, which made the next goslings have natural passive immunity, so the disease would not occur in the same area for two consecutive years. Therefore, the morbidity and mortality of the disease are related to the age of day, on the other hand, it depends on the specific antibody level of the parent goose. Immune prevention is an important measure to control goose plague. At present, PCR,ELISA, indirect immunofluorescence technique and micro neutralization test have been established to detect Gosling plague. However, these methods all need special instruments and equipment, and take a long time, so they are not suitable for popularization and application in basic level. Therefore, it is urgent to establish a more rapid and sensitive method for virus detection. The polymerase chain reaction established in this study has the characteristics of simple operation, fast operation and suitable for basic level popularization. In recent years, Gosling plague has been prevalent in some provinces, cities and autonomous regions of China, which has caused serious harm to the industry of feeding and feeding, and at the same time has brought huge economic losses to the breeders, so it has been widely concerned by scholars and workers at home and abroad. In this study, the current situation of goose breeding and the incidence of Gosling plague in the region were investigated, and the epidemic situation was summarized, in order to make relevant preventive measures to the occurrence of goose plague in the goose industry. In this study, a pair of specific primers were designed based on the sequence information of vp3 of Gosling plague published on GENBANK. By optimizing various conditions, the nucleic acid fragments of 314bp with the expected size were amplified. It was found that the primer could only amplify the specific fragment of Gosling plague and be used to detect the occurrence of Gosling plague. In this study, 10 isolates were sequenced. The results showed that the highest homology was 100% and the lowest was over 95.2% among the isolates, indicating that the nucleotide variation of the amplified fragments of Gosling plague virus was very small, and VP3 was the main antigen of Gosling plague. Therefore, this study has instructive significance for immune prevention. According to the basic information of Gosling plague in this study, especially the immune prevention program in every goose farm, it was found that the incidence age of Gosling plague was between December and February, and according to the incidence of Gosling plague, The results showed that the incidence probability of Gosling plague had been reduced, which indicated that the proper immunization procedure was an effective measure to prevent the disease.
【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.33
本文編號：2285013
[Abstract]:Gosling plague is an acute infectious disease caused by goose parvovirus, which mainly infects goslings and young muscovy ducks within 30 days of age. It is characterized by rapid onset, high mortality and rapid transmission. The morbidity and mortality decreased with the increase of the age of the day, mainly affecting the goslings and young muscovy ducks within 7 days of age, which were divided into three types according to the incidence, the most acute type, the acute type and the subacute type (Yin Zhen and Liu Jinghua, 1997). In 1966, Hungarian scholar Derszy also isolated the virus from goose embryos. He was the first person in foreign countries to isolate the virus from goose embryos, and described the disease in detail in 1967. But at the time he mistook the disease for the flu, It was not until 1971 that Schettler determined that the disease was caused by parvovirus. In 1978, it was suggested that the disease should be called goose parvovirus infection. In 1974, the World Avian Society named the disease Derzsy's disease (Derzsy Dl1975) to commemorate the advance work of Derzsy. Zadori et al. The entire GPV gene sequence, The gene sequence of muscovy duck parvovirus was compared with that of muscovy duck parvovirus (Zadori ZJ 1995). The epidemic has a certain periodicity. After the pandemic, the remaining geese got active immunity, which made the next goslings have natural passive immunity, so the disease would not occur in the same area for two consecutive years. Therefore, the morbidity and mortality of the disease are related to the age of day, on the other hand, it depends on the specific antibody level of the parent goose. Immune prevention is an important measure to control goose plague. At present, PCR,ELISA, indirect immunofluorescence technique and micro neutralization test have been established to detect Gosling plague. However, these methods all need special instruments and equipment, and take a long time, so they are not suitable for popularization and application in basic level. Therefore, it is urgent to establish a more rapid and sensitive method for virus detection. The polymerase chain reaction established in this study has the characteristics of simple operation, fast operation and suitable for basic level popularization. In recent years, Gosling plague has been prevalent in some provinces, cities and autonomous regions of China, which has caused serious harm to the industry of feeding and feeding, and at the same time has brought huge economic losses to the breeders, so it has been widely concerned by scholars and workers at home and abroad. In this study, the current situation of goose breeding and the incidence of Gosling plague in the region were investigated, and the epidemic situation was summarized, in order to make relevant preventive measures to the occurrence of goose plague in the goose industry. In this study, a pair of specific primers were designed based on the sequence information of vp3 of Gosling plague published on GENBANK. By optimizing various conditions, the nucleic acid fragments of 314bp with the expected size were amplified. It was found that the primer could only amplify the specific fragment of Gosling plague and be used to detect the occurrence of Gosling plague. In this study, 10 isolates were sequenced. The results showed that the highest homology was 100% and the lowest was over 95.2% among the isolates, indicating that the nucleotide variation of the amplified fragments of Gosling plague virus was very small, and VP3 was the main antigen of Gosling plague. Therefore, this study has instructive significance for immune prevention. According to the basic information of Gosling plague in this study, especially the immune prevention program in every goose farm, it was found that the incidence age of Gosling plague was between December and February, and according to the incidence of Gosling plague, The results showed that the incidence probability of Gosling plague had been reduced, which indicated that the proper immunization procedure was an effective measure to prevent the disease.
【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.33
【參考文獻】
相關(guān)期刊論文 前10條
1 郭卉;涂飛;呂亞楠;史榮華;金文杰;邵紅霞;錢琨;秦愛建;;抗鵝細小病毒單克隆抗體的制備及其免疫熒光檢測方法的建立[J];中國動物傳染病學報;2013年04期
2 朱小麗;陳少鶯;程曉霞;林鋒強;陳仕龍;王劭;李兆龍;王燕玲;;番鴨小鵝瘟病直接熒光診斷試劑的研制[J];畜牧獸醫(yī)雜志;2012年04期
3 呂雪峰;李忠勛;李志慧;吉鳳濤;蘇雙;;小鵝瘟免疫熒光抗體的制備及檢測方法研究[J];中國動物保健;2012年02期
4 趙文婧;魯承;杜秋明;高旭;高建偉;申峰勇;張穎;袁娜;房靖添;;小鼠抗鵝細小病毒VP3蛋白單克隆抗體的制備[J];動物醫(yī)學進展;2011年12期
5 鮮思美;文心田;曹三杰;黃小波;;鵝副黏病毒和鵝細小病毒復合PCR檢測方法的建立[J];黑龍江畜牧獸醫(yī);2011年21期
6 董浩;徐楠楠;;鵝細小病毒熒光定量PCR檢測方法的建立及應用[J];中國獸藥雜志;2011年09期
7 杜秋明;魯承;王暖成;賈文影;趙文婧;申峰勇;;鵝細小病毒單抗雙夾心ELISA檢測方法的建立[J];中國獸醫(yī)學報;2011年03期
8 朱德康;黎敏;車茜;程安春;汪銘書;陳孝躍;;小鵝瘟病毒VP3真核表達質(zhì)粒與弱毒疫苗誘導鵝體免疫應答的比較[J];中國農(nóng)業(yè)科學;2011年03期
9 李干洲;王璇;徐景娥;余波;楊粵黔;汪德生;;平壩灰鵝群鵝細小病毒病流行病學調(diào)查[J];畜牧與獸醫(yī);2010年12期
10 尚緒增;張雅為;王兆鵬;鞠環(huán)宇;馬波;王君偉;;鵝細小病毒NS2蛋白間接ELISA方法的建立[J];中國預防獸醫(yī)學報;2010年08期
，本文編號：2285013
本文鏈接：http://sikaile.net/yixuelunwen/dongwuyixue/2285013.html
最近更新
教材專著
