本文選題：內(nèi)羅畢羊病毒 + 蜱組織��； 參考：《吉林農(nóng)業(yè)大學》2015年碩士論文

【摘要】：眾所周知,蜱是一種寄生在動物體表的吸血寄生蟲,是多種人畜共患傳染病的重要傳播媒介。蜱蟲分類上屬于動物界節(jié)肢動物門、蛛形綱、螨亞綱、寄螨目、蜱總科。全世界已經(jīng)發(fā)現(xiàn)18個屬897種,而中國已發(fā)現(xiàn)10屬,119種。蜱可引起多種重要的自然疫源性疾病和人獸共患病,如Q熱、發(fā)熱伴血小板減少綜合征、森林腦炎、出血熱等。內(nèi)羅畢羊病毒(Nairobi sheep disease virus)在分類上屬于布尼亞病毒科。其主要傳播媒介是蜱蟲,對公眾危害較為廣泛,能夠引發(fā)急性出血性胃腸炎,綿羊和山羊的死亡率高達90%。該病于1910年在肯尼亞的內(nèi)羅畢附近首次發(fā)現(xiàn),并在1917年確定其病原為內(nèi)羅畢羊病毒。1994年首次報道發(fā)現(xiàn)人感染內(nèi)羅畢羊病毒發(fā)病。內(nèi)羅畢羊病主要發(fā)生在非洲東部的肯尼亞、烏干達、坦桑尼亞、埃塞爾比亞和索馬里。目前,內(nèi)羅畢羊病尚未在中國乃至東亞報道過,因此,對中國東北部內(nèi)羅畢羊病毒(NSDV)進行鑒定及全基因組序列分析具有重要意義�；谀壳皷|北地區(qū)蜱類分布不清、病毒本底尚不明確等因素,本研究在吉林省的集安、敬信、春化、南山、松林,黑龍江省的哈爾濱、呼瑪和遼寧省的丹東等10余個縣市進行了蜱類的采集。通過形態(tài)學鑒定,主要發(fā)現(xiàn)四個蜱蟲物種,豐富的是長角血蜱屬longicornis(84.8%),其次是矩頭蟬屬silvarum(7.2%),矩頭蟬屬nuttalli(5.5%)和Ixodes persulcatus(2.5%)。通過病毒宏基因組學分析,共獲得26411條reads,進一步拼接得到平均長度為108堿基的18997條重疊序列。其中,動物性病毒科數(shù)量最多,占病毒序列總數(shù)的65%(7505/11547),包括:細小病毒科(Parvoviridae),黃病毒科(Flaviviridae),布尼亞病毒科(Bunyaviridae)等;植物性病毒占19%(2189/11547);噬菌體序列占16%(1853/11547)。本研究進一步對蜱組織進行裂解,采用RT-PCR手段再次對內(nèi)羅畢羊病毒(NSDV)進行驗證,結(jié)果得出吉林集安、敬信及遼寧丹東地區(qū)蜱蟲均攜帶內(nèi)羅畢羊病毒。通過以病毒宏基因組學分析數(shù)據(jù)及國內(nèi)外NSDVs病毒序列為參考,本研究成功克隆了內(nèi)羅畢羊病毒(NSDV)的全基因組。該序列與國外NCBI已收錄的全基因組序列進行核苷酸同源性比較,基因片段序列比較顯示:在核苷酸水平上相比于其他NSDVs同源性為75.1~89.6%之間,推導出氨基酸同源性在81.3~96.7%之間。相比于其他物種的成員NSDVs(Dugbe,Kupe,Hazara and Crimean Congo hemorrhagic fever viruses),同源性在37.5-68.6%之間。本研究在東亞地區(qū)首次證實了蜱傳內(nèi)羅畢羊病毒的存在。另外,基于對以上陽性地區(qū)蜱蟲形態(tài)學鑒定,首次發(fā)現(xiàn)了長角血蜱可攜帶內(nèi)羅畢羊病毒,同時為開展東北地區(qū)內(nèi)羅畢羊病毒(NSDV)致病性及疫苗的研究奠定了良好的基礎。
[Abstract]:It is well known that tick is a blood-sucking parasite parasitic on animal surface and an important transmission medium of zoonotic infectious diseases.Ticks are classified as Arthropoda, Arachne, Acari, Acaridae, Acaridae.There are 897 species of 18 genera in the world, while 119 species of 10 genera have been found in China.Ticks can cause many important natural diseases and zoonosis, such as Q fever, fever with thrombocytopenia syndrome, forest encephalitis, hemorrhagic fever and so on.Nairobi sheep disease virus belongs to the Bunia virus family.Its main transmission vector is ticks, which are widely harmful to the public and can cause acute hemorrhagic gastroenteritis. The mortality rate of sheep and goats is as high as 90%.The disease was first found near Nairobi, Kenya, in 1910, and was identified as Nairobi sheep virus in 1917. It was first reported in 1994 that people were infected with Nairobi sheep virus.Nairobi sheep disease mainly occurs in eastern Africa Kenya, Uganda, Tanzania, Ethiopia and Somalia.At present, Nairobi sheep disease has not been reported in China or even in East Asia, so it is of great significance to identify and analyze the whole genome sequence of Nairobi sheep virus (NSDV) in northeastern China.Due to the unclear distribution of ticks and the unclear background of the virus in Northeast China, this study was conducted in Ji'an, Jingxin, Chunhua, Nanshan, Pine Forest and Harbin, Heilongjiang Province.More than 10 counties and cities, such as Huma and Dandong of Liaoning Province, collected ticks.By morphological identification, four species of ticks were found, which were abundant in the genus Haemaphysalis longicornis84.8, followed by Silvarum 7.2dicum, nuttallius 5.5) and Ixodes persulcatus 2.5.A total of 26411 readswere obtained by virus macrogenomics analysis, and 18997 overlapping sequences with an average length of 108bp were obtained by further splicing.Among them, the animal viridae has the largest number, accounting for 65% of the total number of virus sequences, including Parvoviridaeae, Flaviviridaeae, Bunyaviridaeae and so on; plant viruses account for 191899 / 11547; bacteriophage sequences account for 161853 / 11547; phage sequences account for 161853 / 11547.In this study, the tissues of ticks were further cracked, and the Nairobi sheep virus (NSDV) was verified again by RT-PCR. The results showed that Jilin Ji'an, Jingxin and Dandong of Liaoning all carried Nairobi sheep virus.The whole genome of Nairobi Sheep virus (NSDV) was successfully cloned by reference to the data of viral macrogenomics analysis and the sequences of NSDVs virus at home and abroad.The nucleotide homology of this sequence was compared with that of the whole genome sequence of foreign NCBI. Compared with other NSDVs, the nucleotide homology of this sequence was 75.1% and the deduced amino acid homology was 81.33% and 96.7% respectively.The homology of Hazara and Crimean Congo hemorrhagic fever virusesus was 37.5-68.6% compared with other species.In this study, the existence of tick-borne Nairobi sheep virus was first confirmed in East Asia.In addition, based on the morphological identification of the above positive ticks, Haemaphysalis longicornis was found to carry the Nairobi sheep virus for the first time, which laid a good foundation for the study of the pathogenicity and vaccine of NSDV in Northeast China.
【學位授予單位】：吉林農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S852.654

【參考文獻】

相關期刊論文 前3條

1 沈愛華;鮑昌俊;胡建利;李陸遜;羊海濤;;6例人感染新型布尼亞病毒病的臨床特征和流行病學調(diào)查分析[J];東南大學學報(醫(yī)學版);2012年05期

2 ZHANG XiaoShuang;LIU Yan;ZHAO Li;LI Bing;YU Hao;WEN HongLing;YU Xue-Jie;;An emerging hemorrhagic fever in China caused by a novel bunyavirus SFTSV[J];Science China(Life Sciences);2013年08期

3 張晶波,溫博海,陳梅玲,邱玲,牛東升,李麗莉;檢測貝氏柯克斯體的實時熒光定量PCR[J];中國人獸共患病雜志;2005年08期

，

本文編號：1767053