發(fā)布時間：2017-12-31 20:13

  本文關(guān)鍵詞：湘瓊兩地蝙蝠攜帶星狀病毒與諾如病毒的調(diào)查研究　出處：《南方醫(yī)科大學》2011年碩士論文　論文類型：學位論文

  更多相關(guān)文章： 蝙蝠 諾如病毒 星狀病毒 進化樹分析 檢出率

【摘要】：研究背景和目的 研究表明,蝙蝠是多種人獸共患疾病病毒的自然宿主,并可以跨越物種間屏障,將病毒傳播給人類和其他動物。目前研究人員已經(jīng)從蝙蝠身上發(fā)現(xiàn)或分離很多種病毒,約有18個病毒科包含70多種病毒。已知能從蝙蝠傳播給人類的病毒主要有狂犬病毒、尼巴病毒、亨德拉病毒,還有類SARS冠狀病毒,這些病毒都可以引起人類致命性的疾病。其他病毒,如甲病毒屬、黃病毒屬和布尼亞病毒屬病毒可通過節(jié)肢動物傳播給蝙蝠,但還不確定蝙蝠是不是這類病毒所致疾病的播散宿主�？梢�,還需要大量研究,以明確蝙蝠在病毒性自然疫源性疾病中所扮演的角色。 我國約有120種蝙蝠,但國內(nèi)對蝙蝠的研究工作起步較晚,對其生態(tài)學、行為學研究尚不夠深入,尤其是對我國蝙蝠攜帶哪些種類病毒,以及病毒與蝙蝠生態(tài)學的關(guān)系,對人類健康的影響的研究還較少。迄今為止,我國科研工作者對蝙蝠所攜帶病毒的研究有以下發(fā)現(xiàn)：從蝙蝠體內(nèi)分離到羅斯河病毒、乙型腦炎病毒、基孔肯雅病毒、森林腦炎病毒、SARS類冠狀病毒、狂犬病病毒、星狀病毒、尼巴病毒等。這些發(fā)現(xiàn)表明我國蝙蝠可攜帶某些我國常見傳染病的病毒。 病毒性胃腸炎(viral gastroenteritis)又稱病毒性腹瀉,是一組由多種病毒引起的急性腸道傳染病。臨床特點為起病急、惡心、嘔吐、腹痛、腹瀉,排水樣便或稀便,也可有發(fā)熱及全身不適等癥狀,病程短,病死率低。病毒性腹瀉在兒童中更是一種常見病和多發(fā)病,是導致兒童死亡的重要原因。引起兒童腹瀉的病毒主要有輪狀病毒(rotavirus)、腸道腺病毒(adenovirus)、星狀病毒(astrovirus, Astv)、諾如病毒(norovirus, NoV)等。 患者與無癥狀帶毒者是病毒性胃腸炎的主要傳染源。糞-口途徑傳播是病毒性胃腸炎最重要的傳播途徑,少部分可以通過空氣傳播,其可通過被污染的食物或水引起人群胃腸炎暴發(fā)或散發(fā)�；仡櫳鲜兰o90年代的亨德拉病毒和尼帕病毒的暴發(fā)事件,也是源于食源性感染,是由于蝙蝠病毒污染了水域、水果,再依次傳播給馬、豬、人,最后導致疾病暴發(fā)。 星狀病毒(Astrovirus, Astv)于1975年在胃腸炎患兒糞便標本中首次發(fā)現(xiàn),由于電鏡下病毒顆粒呈星形,因此被命名為星狀病毒。此后世界各地均已有星狀病毒感染的報道,它既可散發(fā)也可以引起暴發(fā)流行,亦可引起醫(yī)源性感染。住院腹瀉患兒中星狀病毒檢出率為2.5%-9.0%,目前認為星狀病毒是嬰幼兒病毒性胃腸炎的第二位病因,僅次于輪狀病毒。 星狀病毒既可感染人,也可感染牛、羊、豬、狗、鹿、火雞、鴨、鼠等多種動物,導致不同程度的胃腸炎。人群感染的星狀病毒,傳統(tǒng)上可分為8個血清型。最近,研究者又在腹瀉病人中發(fā)現(xiàn)兩種的新星狀病毒,astrovirus MLB和astrovirus VA。研究還發(fā)現(xiàn),Astv MLB 1與新近發(fā)現(xiàn)的鼠星狀病毒很相近;Astv VA1在進化樹上與感染羊和貂的星狀病毒最為接近。這提示新發(fā)現(xiàn)的人星狀病毒可能和動物的星狀病毒存在一定的關(guān)系。 近年來,香港大學研究小組分別從香港和中國大陸的蝙蝠體內(nèi)檢出星狀病毒,檢出率高達46%和44.8%；基因分析顯示蝙蝠星狀病毒具有高度基因多態(tài)性,部分病毒可能與感染人的星狀病毒有系統(tǒng)進化的關(guān)系。然而關(guān)于蝙蝠星狀病毒的生態(tài)學資料還很稀少,有必要進一步擴充不同地區(qū)和不同種類的蝙蝠樣本進行研究。 諾如病毒(Norovirus, NoV)是一組杯狀病毒屬病毒,其原型株諾瓦克病毒于1972年在美國諾瓦克市被分離發(fā)現(xiàn)。由于該組病毒極易變異,此后在其他地區(qū)又相繼發(fā)現(xiàn)并命名了多種類似病毒,統(tǒng)稱為諾如病毒。NoV是一種高致病的、傳染性極強的腸胃病毒,能導致急性胃腸炎,一旦有人感染,通常會發(fā)展為群體性的大規(guī)模傳染。遺憾的是,由于病毒的高度變異性及其缺少合適的動物研究模型,現(xiàn)今還未研發(fā)出理想的NoV疫苗,患者容易發(fā)生反復感染。 自然界中攜帶NoV的物種很多,包括有人、豬、牛、鼠、海獅等,可導致它們發(fā)生不同程度的癥狀。根據(jù)基因特征,NoV被分為五個遺傳群(Gene Group, GG):GGⅠ、GGⅡ、GGⅢ、GGⅣ和GGV。五個遺傳群中,GGⅠ、GGⅡ和GGIV主要感染人類,GGⅢ感染牛,GGV則在鼠類動物內(nèi)檢出。 雖然研究已表明NoV在多種動物體內(nèi)廣泛存在,但是關(guān)于動物NoV感染的流行病學還不太清楚。動物NoV在動物體內(nèi)感染率差別較大,研究發(fā)現(xiàn)北美的實驗鼠鼠類NoV的感染率高達22.1%,英國檢測發(fā)現(xiàn)牛的腹瀉病例中有11%比例為NoV感染所致,而豬NoV檢出率比較低,在日本豬GGⅡNoV的檢測率僅為0.35%,在荷蘭檢測率為2.0%。 盡管現(xiàn)今沒有證據(jù)表明動物NoV可以感染人類,但是分子生物學分析表明動物NoV和人類NoV的毒株相互之間有密切的關(guān)系,尤其是豬NoV的毒株可被歸入到與人類NoV毒株相同的基因群(GGⅡ)。新近研究發(fā)現(xiàn)豬體內(nèi)普遍存在人NoV的抗體,這進一步加強了動物可能充當著人類NoV儲存庫的假設。此外,研究還發(fā)現(xiàn)貝殼類動物可同時攜帶人類NoV和動物NoV,這提示一種新的基因重組株可能出現(xiàn)。最近,在加拿大人們檢測到豬NoV和牛NoV的序列與人NoVGGⅡ.4毒株的序列非常接近。因此,將來或許要重新評估NoV種間傳播的可能性及其人畜共患的風險。 2009年,武漢病毒研究所在蝙蝠體內(nèi)檢測到另一種導致人腹瀉的病毒—腸道腺病毒,檢出率為8%(28/350),基因分析顯示蝙蝠腺病毒亦具有高度基因多樣性,這提示蝙蝠可能有攜帶一系列高變異病毒的能力。研究已經(jīng)在蝙蝠體內(nèi)發(fā)現(xiàn)了星狀病毒和腺病毒兩種腹瀉相關(guān)病毒,關(guān)于蝙蝠是否攜帶有NoV,暫未見有文獻報道,因而我們不禁疑問蝙蝠是否攜帶有可導致爆發(fā)流行的NoV。鑒于NoV的高度基因變異性和多宿主性,作為種類第二多的哺乳動物(1000多種)一蝙蝠也很有可能攜帶此類病毒,甚至可能是它們的宿主。 因此,為進一步了解不同地區(qū)不同種類蝙蝠攜帶星狀病毒的本底情況,探討其與感染人星狀病毒的關(guān)系,同時了解這些地區(qū)的蝙蝠是否攜帶諾如病毒,本研究擬在先前未見報道的湖南、海南部分地區(qū)為現(xiàn)場,對當?shù)仳饠y帶星狀病毒和諾如病毒情況進行流行病學調(diào)查。 研究方法 1.樣本的采集和處理 2007年7月至2008年7月,于岳陽、邵陽、�？谌貐^(qū)蝙蝠長期監(jiān)測點中選取4個蝙蝠自然棲息地,現(xiàn)場捕捉蝙蝠,進行編號后,請蝙蝠研究專家進行鑒定。標本采集方式分兩種,分別為采集肛拭子和剪取直腸組織。采集的樣本存放于含200μl RNAlater儲存液的凍存管中,置放于密封的冰袋泡沫盒中當天運回實驗室,放于-80℃冰箱貯存,待檢。 2.蝙蝠病毒的檢測與鑒定2.1星狀病毒 ①半巢式PCR篩檢樣本 參照文獻,針對星狀病毒高度保守的RdRp基因,設計合成5條用于巢式PCR的引物,包括4條前引物和1條共同的后引物,應用巢式PCR方法檢測標本。 ②序列鑒定與分析 對陽性PCR產(chǎn)物進行切膠、純化和回收,然后送樣至公司測序。如遇陽性PCR產(chǎn)物條帶很弱者,則先做克隆,再取接種培養(yǎng)后菌液送樣至公司測序鑒定。 ③將測序結(jié)果導入GenBank的BLAST軟件進行在線同源性分析,然后在GenBank下載相關(guān)核酸序列,利用Clastal_X軟件對所要分析的序列進行多序列比對分析,并用DNAstar軟件對核酸序列相似性進行計算。使用MEGA4.1軟件,用Neighbor-Joining法構(gòu)建進化樹進行分析。 2.2諾如病毒 ①TaqMan real-time PCR檢測基因組Ⅱ型諾如病毒(GGⅡNoV) 參照文獻并合成用于檢測GGⅡNoV公認的特異引物和探針,利用TaqMan real-time PCR方法對本進行檢測。 ②SYBR Green real-time PCR檢測諾如病毒 參照文獻合成可檢測NoV的兩對通用引物P289/P290和12Y/13I,應用SYBR Green real-time PCR方法分別檢測標本。 ③GGV型NoV特異引物檢測標本 參照文獻合成公認的可檢測GGV型NoV引物,應用普通RT-PCR方法檢測標本。 ④對普通RT-PCR擴增獲得的可疑陽性PCR產(chǎn)物進行切膠、純化和回收,然后送樣至公司測序。對TaqMan real-time PCR檢測的可疑陽性PCR產(chǎn)物,送樣至公司克隆鑒定。 3.質(zhì)量控制 ①移液槍頭、凍存管、EP管等實驗耗材均為一次性用品。 ②RNA提取和逆轉(zhuǎn)錄實驗所采用的所有容器和配制試劑用DEPC水處理,高溫高壓處理,烘干待用,以去除RNA酶污染。 ③全程戴口罩、手套、帽子,在無菌室生物安全柜進行操作,防止試劑和實驗過程受到污染。 ④實驗過程嚴格設立陽性對照、陰性對照和空白對照。 結(jié)果 1蝙蝠基本情況 本次研究在湖南、海南兩省四個監(jiān)測點共采集蝙蝠321只,涉及2個蝙蝠亞目4個蝙蝠科9個種類的蝙蝠。共制備蝙蝠糞便標本321份,包括肛拭子59份和直腸樣本262份。 2星狀病毒的檢測 ①巢式RT-PCR檢測到陽性標本29例,總陽性率為9.03%(29/321),在9種表面健康的蝙蝠中發(fā)現(xiàn)有3種蝙蝠攜帶有星狀病毒,陽性蝙蝠種類包括長翼蝠(22/187)、小黃蝠(6/38)和棕果蝠(1/59),以小黃蝠的檢出率最高。 ②以星狀病毒RdRp基因部分氨基酸序列構(gòu)建的進化樹分析顯示,本研究檢測的蝙蝠星狀病毒不僅與來源于其他種類的星狀病毒差異較大,而且本身也有很大的差異。然而,有5個蝙蝠星狀病毒卻與羊星狀病毒(ovine Astv)、貂星狀病毒(mink Astv)和人新型星狀病毒Astv VA在進化樹上最為相近,序列相似性為51.6%-63.1%。另外,有8株從邵陽地區(qū)同一個洞穴長翼蝠檢測到的星狀病毒相聚在一小分支,它們的氨基酸序列相似性為97%-99%。 3諾如病毒的檢測 本研究采用TaqMan real-time PCR、SYBR Green real-time PCR和普通RT-PCR三種實驗方法,應用多對引物分別篩查上述蝙蝠糞便樣本GGⅠ-Ⅳ型NoV攜帶情況,檢測結(jié)果均為陰性。 結(jié)論 ①巢式PCR檢測到星狀病毒陽性率為9.03%(29/321),并首次在果蝠身上檢出星狀病毒(1/59),提示此類星狀病毒可能適應于翼手目動物,盡管不同種類的蝙蝠對它的易感程度不一。 ②本研究檢出的蝙蝠星狀病毒基因多態(tài)性豐富,這結(jié)果與先前研究一致。5株蝙蝠星狀病毒卻與羊星狀病毒、貂星狀病毒、人星狀病毒Astv VA在進化樹上最為相近,并具有較高的氨基酸序列相似性。這提示蝙蝠星狀病毒可能跨種族傳播給人類或其他動物,亦或是在它們之間相互傳播。 ③來自邵陽同一個洞穴檢測得到的8株星狀病毒具有高度的基因相似性,進一步提示蝙蝠星狀病毒在同一個洞穴內(nèi)傳播的可能。 ④本研究從所收集的321份蝙蝠糞便樣本中未檢測到諾如病毒,提示蝙蝠攜帶諾如病毒的可能性不大。由于諾如病毒高度遺傳變異,而且現(xiàn)今沒有一對單獨引物可同時檢測出全部五個基因組的諾如病毒。蝙蝠是否攜帶諾如病毒還不清楚,有待進一步研究。
[Abstract]:Background and purpose of research
Research shows that bats are many zoonotic virus natural host, and can cross the species barrier and spread the virus to humans and other animal. At present, researchers have found that separation from the bats or many types of viruses, about 18 virus, including more than 70 kinds of virus. Known to spread to humans from bats the virus mainly has the rabies virus, Nipah virus, Hendra virus, and SARS coronavirus, these viruses can cause human fatal disease. Other viruses, such as a virus, and the virus flavivirus bunyavirus by arthropod animal spread to bat, but it is not clear whether bats are of this kind of disease caused by virus the spread of the host. Obviously, we still need a lot of research, in order to clear the bat in viral disease of natural focus in the role.
There are about 120 species of bats in China, but the domestic research work on bat started late, the ecology, behavioral research is still not enough, especially on what kind of virus carrying our bat, the relationship between virus and bat ecology, study the impact on human health is less. So far, the research in our country scientists are carrying the virus of bats are as follows: from separation to bat in the Ross River virus, Japanese encephalitis virus, chikungunya virus, Semliki Forest virus, SARS coronavirus, rabies virus, astrovirus, Nipah virus. These findings suggest that China's bat can carry some of common infectious diseases in China virus.
Viral gastroenteritis (viral gastroenteritis) also known as viral diarrhea, is a group of acute intestinal infectious disease caused by a variety of viruses. The clinical features of acute onset, nausea, vomiting, abdominal pain, diarrhea, drain or rare, also can have fever and malaise and other symptoms, short duration, low fatality rate. Viral diarrhea in children is a common disease, is an important cause of death in children. Children's diarrhea virus mainly caused by rotavirus (rotavirus), enteric adenovirus (adenovirus), astrovirus (astrovirus, Astv), norovirus (norovirus, NoV)..
Patients with asymptomatic carriers is the main source of infectious viral gastroenteritis. The fecal oral route transmission is viral gastroenteritis is the most important part, can be spread through the air, it can be through contaminated food or water caused by the crowd or sporadic events. An outbreak of gastroenteritis outbreaks were Hendra and Nipah viruses in the world in 90s, also due to foodborne infection, is due to bat virus contaminated water, fruit, then spread to the horse, pig, human, and finally lead to disease outbreaks.
Astrovirus (Astrovirus, Astv) in 1975 for the first time found in the stool samples of children with gastroenteritis, because the virus particles under the electron microscope is the star, it is named astrovirus. Then all over the world have astrovirus infection reported, it can also cause sporadic epidemic of violence, can also cause iatrogenic infection hospitalized children with diarrhea. Astrovirus detection rate was 2.5%-9.0%, the astrovirus is infant viral gastroenteritis second cause, second only to rotavirus.
Astrovirus can infect people, but also can infect cattle, sheep, pigs, dogs, deer, turkeys, ducks, rats and other animal, lead to different degrees of gastroenteritis. Astrovirus infection, traditionally can be divided into 8 serotypes. Recently, researchers have found that two kinds of star shaped virus in diarrhea patients, astrovirus MLB and astrovirus VA., the study also found that Astv and MLB 1 were astrovirus newly found very similar; Astv VA1 in evolutionary tree with infected sheep and mink astrovirus is most close. There is a certain relationship between the stellate virus suggesting human astrovirus and may the newly discovered animal.
In recent years, were detected astrovirus from Hongkong and Chinese, the bat in the University of Hong Kong research team, the detection rate of 46% and 44.8%; gene analysis showed that bat astrovirus has a high degree of polymorphism, relationship between system of astrovirus infection with the virus may some people. However ecological information about bats star the shape of the virus is still scarce, it is necessary to further expand and different kinds of bats samples in different regions.
Norovirus (Norovirus, NoV) is a group of calicivirus is a virus, the prototype strains of Novak virus in the United States in 1972, Novak was found. Because of this group of viruses isolated easily after variation, in other areas have been discovered and named several similar viruses, collectively referred to as norovirus is a highly pathogenic.NoV the highly contagious stomach virus, can cause acute gastroenteritis, once a person is infected, usually for the development of large-scale infectious group. Unfortunately, due to the high variability of the virus and the lack of suitable animal model, now has not yet developed the ideal NoV vaccine, patients prone to recurrent infections.
The nature of NoV carrying many species, including human, pig, cattle, rat, sea lions, they can lead to different degrees of symptoms. According to the characteristics of NoV gene, were divided into five genetic groups (Gene Group, GG): GG 1, GG II, GG III, GG I GG IV and GGV. five genetic groups, GG, 2 and GGIV can infect humans, GG infection in cattle, GGV is detected in rodent animal.
Although studies have shown that NoV is widely present in a variety of animal body, but epidemiological about animal NoV infection is not clear. NoV in different animal rate in infected animal, experimental study found that mouse class NoV North America infection rate as high as 22.1%, the British found cattle in 11% cases of diarrhea caused by NoV infection ratio but, pig NoV detection rate is relatively low, in the detection of Japanese pig GG II NoV rate was only 0.35% in Holland, the detection rate was 2.0%.
Although there is no evidence that the animal NoV can infect humans, but molecular analysis revealed that NoV animal and human NoV strains were closely related to each other, especially the swine NoV strain can be classified into the same group of genes with human NoV strains (GG II). Recent studies have found that the antibody against porcine is common in human NoV this, to further strengthen the animal may play the role of a human NoV storage hypothesis. In addition, the study also found that shellfish animal can simultaneously carry human and animal NoV NoV, suggesting a possible new gene recombinant strains. Recently, people in Canada are very close to the sequence detected in porcine NoV and bovine NoV sequences and NoVGG II.4 strain. Therefore, the future may have to re evaluate the risk possibility of the spread of NoV species and zoonoses.
In 2009, Wuhan virus Institute bats detected another cause diarrhea virus, enteric adenovirus, the detection rate was 8% (28/350), gene analysis showed that bat adenovirus also has high genetic diversity, suggesting that bats may have the ability to carry a series of highly variable virus research has found the star. Rotavirus and adenovirus two diarrhea related virus in bats, a bat is carrying NoV, yet see the reports in the literature, so we can not help but doubt whether bats carrying a outbreak of NoV. in height based NoV due to variability and multiple hosts, as the more than 2 species of mammals (1000 kinds) a bat is likely to carry the virus, or even their host.
Therefore, in order to further understand the different regions of different types of bats astrovirus the situation at the end of its relationship with human astrovirus infection, while understanding in these areas are carrying bats norovirus, this study has not been reported previously in Hunan, parts of Hainan for the scene, the local bats in the investigation of astrovirus and norovirus.
research method
Collection and processing of 1. samples
From July 2007 to July 2008, in Yueyang, Shaoyang, Haikou three areas of long-term monitoring of bats from 4 bats natural habitat in the scene to capture the bat, number, please bat research experts were identified. Samples were collected two hours, including rectal swab samples and cut from rectum tissue samples stored in water. 200 L RNAlater storage solution in freezing tube, placed in the ice box sealing foam when in the day back to the lab, put in the -80 C refrigerator storage, to be inspected.
Detection and identification of 2. bats virus 2.1 stellate viruses
1. Semi nested PCR screening sample
Referring to the literature, for the highly conserved RdRp gene of the stellate virus, we designed and synthesized 5 primers for nested PCR, including 4 primers and 1 common primers. Nested PCR was used to detect the samples.
Sequence identification and analysis
The positive PCR products were excised, purified and recovered, and then sent to the company for sequencing. In case of positive PCR products with very weak, then do cloning, then after inoculation of bacteria feeding to DNA sequencing.
The sequencing results into GenBank BLAST software for analysis of online source, and then download the related nucleic acid sequences in GenBank, use Clastal_X software to analyze the multiple sequence alignment of sequences to be analyzed, and the nucleic acid sequence similarity was calculated by DNAstar software. Using MEGA4.1 software, were analyzed by Neighbor-Joining method to construct phylogenetic trees.
2.2 norovirus
(1) TaqMan real-time PCR detection of geno type Nuo virus (GG II NoV)
The specific primers and probes used to detect GG II NoV were synthesized with reference to the literature, and the TaqMan real-time PCR method was used to detect the primers.
(2) SYBR Green real-time PCR detection of norovirus
Two pairs of universal primers P289/P290 and 12Y/13I were synthesized with reference to the literature, and the specimens were detected by SYBR Green real-time PCR method respectively.
GGV type NoV specific primer detection specimens
GGV type NoV primers were detected by reference to the literature, and the specimens were detected by the common RT-PCR method.
Fourth, the suspicious and positive PCR products obtained from common RT-PCR were cut, purified, recovered, then sent to the company for sequencing. The suspicious positive PCR products detected by TaqMan real-time PCR were sent to the company for cloning and identification.
3. quality control
(1) the experimental materials such as the liquid gun head, the cryopreservation tube and the EP tube are all disposable supplies.
(2) all the containers and reagents used in the RNA extraction and reverse transcriptase experiment were treated with DEPC water, high temperature and high pressure treatment, and dried to remove RNA enzyme pollution.
3. Wear masks, gloves and hats in the whole process, and operate in the asepsis room biological safety cabinet to prevent the reagent and the experimental process from being polluted.
4. Positive control was strictly set up in the experimental process, negative control and blank control.
Result
1 basic situation of bats
In this study, 321 bat species were collected from four monitoring sites in two provinces of Hunan and Hainan, involving 2 bat species, 4 bat families, 9 species of bats. Altogether 321 specimens of bat feces were prepared, including 59 59 anal swabs and 262 262 rectal samples.
Detection of 2 stellate viruses
The nested RT-PCR detected 29 positive samples, the total positive rate was 9.03% (29/321), in 9 apparently healthy bats were found in 3 species of bats carrying astrovirus, positive bat species including m.schreibersii (22/187), yellow bat (6/38) and brown (1/59), with yellow fruit bats the highest detection rate.
The star virus RdRp gene to construct phylogenetic tree of amino acid sequence part analysis shows that bat astrovirus in this study not only to detect astrovirus differences and from other types of large, but also have great differences. However, there are 5 bat astrovirus and sheep astrovirus (ovine Astv) mink astrovirus (mink, Astv) and a novel human astrovirus Astv VA in evolutionary tree is most similar to the sequence similarity of 51.6%-63.1%. and 8 strains from Shaoyang region in the same cave m.schreibersii detected astrovirus gathered on a small branch, their amino acid sequence similarity to 97%-99%.
Detection of 3 norovirus
In this study, TaqMan real-time PCR, SYBR Green real-time PCR and ordinary RT-PCR were used in three experimental methods. Multiple pairs of primers were used to screen the carrying condition of GG type I - IV NoV in the stool samples.

【學位授予單位】：南方醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R373

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相關(guān)期刊論文 前1條

1 李文東,梁國棟,梁冰,胡志紅,石正麗,張樹義;蝙蝠攜帶病毒的研究進展[J];中國病毒學;2004年04期

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