臺灣博碩士論文加值系統

輪狀病毒是造成世界各地嬰幼兒病毒性急性腸胃炎的首要病因。依據病毒之VP6蛋白質，可將輪狀病毒分成 A-G 七群，目前已知會感染人類之輪狀病毒為A、B、C群。根據國外的研究報告指出，B、C群輪狀病毒主要感染對象為成人，而成人腹瀉時往往不會收集糞便送檢，因此欲研究B、C群輪狀病毒在台灣的感染狀況，在檢體取得上相當不易。
2003年本實驗室於台大醫院的輪狀病毒陽性糞便臨床檢體中，由RNA電泳之核酸分布型態發現了三株C群輪狀病毒。為進一步了解C群輪狀病毒在台灣的感染情況，今利用血清流行病學的方式進行研究。本研究所使用的抗原為利用重組桿狀病毒所表現之輪狀病毒VP6蛋白質，由於桿狀病毒的宿主為無脊椎動物，通常不會與人類血清產生交叉反應，適合用於表現血清學研究時所需的抗原。
為了解C群輪狀病毒抗體陽性率分布狀況，使用酵素連結吸附實驗偵測1376個血清檢體，這些檢體於2006年採取，是隨機選自台北縣市不同地區，並考慮性別以及各年齡層分布之平均，當 cut-off 值為 mean + 2 SD 時，總陽性率為 34.5 %；cut-off 值為 mean + 3 SD 時，總陽性率為 27.2 %，以此兩種 cut-off 值進行分析時，發現： ( 1 ) 隨著年齡層的增加，OD值及陽性率皆有上升的趨勢；( 2 ) 性別與陽性率無顯著相關 ( p = 0.936 )；( 3 ) 人口密度低之地區相較於人口密度高之地區，陽性率較高，且具顯著意義 ( p<0.05 )，顯示都會地區的血清較少存在針對C群輪狀病毒之抗體；( 4 ) 家畜密度高之地區相較於家畜密度低之地區，陽性率較高，且具顯著意義 ( p<0.05 )，顯示血清中針對C群輪狀病毒之抗體存在與否可能與家畜接觸有關。
在篩檢B群輪狀病毒抗體方面，本研究使用的是西方墨點法，隨機選取2006年台北縣市各年齡層及性別分布相近之檢體共 427 個，實驗結果發現只有6個檢體呈現陽性反應，陽性率為 1.41 %。

Rotavirus is one of the most important agents of acute infantile gastroenteritis. Rotaviruses have been classified into seven groups ( A to G ) on the basis of VP6. Group B and C are known to be able to infect human, especially adults. It is difficult to study the epidemiology of group B and C rotaviruses because adults seldom visit doctors and sand their stool samples for diarrheal illness.
In 2003, our laboratory found 3 cases of group C rotavirus, according to RNA PAGE pattern, from National Taiwan University Hospital. In this study, antibodies to group C rotavirus were detected in different populations in Taiwan. Recombinant VP6 expressed from baculovirus was used as the antigen for ELISA test to detect the antibodies to group C rotavirus.
For detection of antibodies to group C rotavirus 1376 serum samples collected in 2006 were random by selected, genders and age groups were evenly distributed. When cut-off value was set as the mean + 2 SD, the antibody positive rate was 34.5 %; when cut-off value was set as the mean + 3 SD, the antibodies positive rate was 27.2 %. We found that ( 1 ) OD values and the positive rates were increasing with age increasing; ( 2 ) there was no apparent difference between male and female ( p = 0.936 ); ( 3 ) the positive rates of regions with density of human population was higher than region with high density of human population ( p < 0.05 ); ( 4 ) the positive rates of region with high density of livestock population was higher than regions with low density of livestock population ( p<0.05 ), perhaps the chance of contact with livestock made the difference.
To screen the antibodies to group B rotavirus, Western blot assay was used. Only 6 samples were positive after detection of 427 serum samples collected from Taipei in 2006, the positive rate was 1.41 %.

口試委員會審定書……………………………………………………………… 2
誌謝……………………………………………………………………………… 3
中文摘要………………………………………………………………………… 5
英文摘要…………………………………………………………………………. 6
第一章 緒論……………………………………………………………………. 13
第一節 輪狀病毒
一、基本結構………………………………………………............ 13
二、病理徵狀……………………………………………………… 13
三、人類輪狀病毒分群…………………………………………..... 14
第二節 偵測人類B群及C群輪狀病毒感染狀況之沿革
一、偵測抗原………………………………………………………. 16
二、偵測抗體………………………………………………………. 17
第三節 桿狀病毒
一、桿狀病毒分群…………………………………………………. 18
二、基本結構……………………………………………………..... 18
三、基因表現……………………………………………………..... 19
四、利用桿狀病毒做為蛋白質表現系統………………………..... 20
第二章 材料與方法…………………………………………………………........ 21
第一節 C型輪狀病毒基因之種系分析
一、檢體來源……………………………………………………….. 22
二、RNA萃取…………………………………………………….... 22
三、反轉錄聚合酶連鎖反應反應………………………………….. 23
四、純化DNA………………………………………………………. 23
五、核酸定序聚合酶連鎖反應………………………………………. 23
六、核酸定序反應物純化……………………………………………. 23
七、核酸定序儀操作………………………………………………… 24
第二節 昆蟲細胞培養
一、培養基配製……………………………………………………… 24
二、細胞繼代培養…………………………………………………….25
三、細胞的冰凍保存………………………………………………….26
第三節 桿狀病毒培養及蛋白質表現
一、偵測病毒力價…………………………………………………….26
二、病毒培養…………………………………………………………27
三、細胞的冰凍保存………………………………………………….28
第四節 蛋白質及抗體偵測
一、蛋白質處理方法………………………………………………….28
二、蛋白質偵測……………………………………………………….30
三、抗體偵測………………………………………………………….33
第三章 結果……………………………………………………………………….. 35
第一節 台大醫院C群輪狀病毒基因序列之分析
一、基因序列分析…………………………………………………….35
二、種系分析………………………………………………………….35
第二節 VP6蛋白質表現偵測
一、重組桿狀病毒之力價測試……………………………………….36
二、重組桿狀病毒表現VP6收集之時間點測試……………………36
三、利用抗體確認VP6蛋白質表現狀況……………………………37
四、利用間接免疫螢光染色來確定 VP6 在細胞的表現狀況……..37
第三節 偵測血清中是否有針對C群輪狀病毒之抗體存在
一、曾感染過C群輪狀病毒之孩童與其家人……………………….38
二、以酵素連結免疫吸附實驗偵測台北縣市各年齡層血清……..…38
三、細胞的冰凍保存…………………………………………………..40
第四節 偵測血清中是否有針對B群輪狀病毒之抗體存在
一、中國大陸恢復期血清之抗體力價偵測…………….…………….40
二、台北縣市隨機血清抗體偵測……………………………………..41
第四章 討論 ……………………………………………………………………......42
參考文獻…………………………………………………………………….…………75
附錄…………………………………………………………………………………….81

圖目錄
圖1. 台大醫院C群輪狀病毒臨床檢體 VR030525、VR030549、
VR030673與可表現C群輪狀病毒VP6之重組桿狀病毒間的
VP6 核酸序列比較…………………………………………………….49
圖2. C群輪狀病毒 NSP4 基因之種系分析……………………………… 50
圖3. C群輪狀病毒VP7 基因之種系分析………………………………… 51
圖4. C群輪狀病毒VP6 基因之種系分析………………………………… 52
圖5. 重組桿狀病毒表現C群輪狀病毒VP6之收集時間點測試………… 53
圖6. 重組桿狀病毒表現B群輪狀病毒VP6之收集時間點測試………… 54
圖7. 使用西方墨點法偵測 HRP-anti ADRV antibody 與重組桿狀病毒
表現A、B、C群輪狀病毒VP6之抗體抗原反應………………… 55
圖8. 使用西方墨點法偵測 rabbit anti rotavirus group C VP6 antibody
與重組桿狀病毒表現A、B、C群輪狀病毒VP6之抗體抗原
反應…………………………………………………………………… 56
圖9. 使用西方墨點法偵測 rabbit anti NCDV rotavirus group A VP6 antibody
與重組桿狀病毒表現A、B、C群輪狀病毒VP6之抗體抗原反應… 57
圖10. 利用間接免疫螢光染色偵測VP6重組蛋白質在細胞中的表現狀況…58
圖11. 以西方墨點法偵測台大醫院三名感染過C群輪狀病毒者及其家人
於 2004 年至 2006 年提供之血清…………………………………… 59
圖12 . 利用間接免疫螢光染色偵測VR030673之姐的血清………………… 60
圖13. 以年齡分群，C群輪狀病毒之陽性率………………………………… 61

表目錄
表1. 針對C群輪狀病毒基因VP6、VP7、NSP4定序反應所使用
之核酸引子……………………………………………………………… 62
表 2. C群輪狀病毒NSP4 基因的核酸及胺基酸序列之比較…………… 63
表 3. C群輪狀病毒VP7 基因的核酸及胺基酸序列之比較……………… 64
表 4. C群輪狀病毒VP6 基因的核酸及胺基酸序列之比較……………… 65
表 5. 台大醫院三名曾感染過C群輪狀病毒孩童與其家人以ELISA
偵測C群輪狀病毒之抗體……………………………………………… 66
表 6. 血清中針對C群輪狀病毒VP6之酵素連結吸附試驗OD值與
西方墨點法對照………………………………………………………… 67
表 7. 年齡分群後使用不同 cut-off 值比較其陽性率……………………… 68
表 8. 東門國小三、四、五年級以不同 cut-off 值比較其陽性率……………69
表 9. 以人口密度分群，比較群組間輪狀病毒之陽性率………………………70
表 10. 人口密度與C群輪狀病毒陽性率之相關性……………………………71
表 11. 家畜密度與C群輪狀病毒陽性率之相關性……………………………72
表 12. 中國大陸感染過B群輪狀病毒恢復期血清，以西方墨點法偵測
針對B 群輪狀病毒VP6之抗體力價………………………………… 73
表 13. 以西方墨點法偵測台北縣市隨機血清樣本中B群輪狀病毒VP6
之抗體陽性率……………………………………………………………74

Flewett, T.H., et al., Relation between viruses from acute gastroenteritis of children and newborn calves. Lancet, 1974. 2(7872): p. 61-3.
2.Kalica, A.R., et al., Comparison of human and animal rotavirus strains by gel electrophoresis of viral RNA. Virology, 1978. 87(2): p. 247-55.
3.KE., M., Rotavirus and their replication. Field B N and Knipe D M 4th ed. Vol. 2. 1747-1786.
4.Clark, S.M., R.S. Spendlove, and B.B. Barnett, Role of two particle types in bovine rotavirus morphogenesis. J Virol, 1980. 34(1): p. 272-6.
5.Vende, P., Z.F. Taraporewala, and J.T. Patton, RNA-binding activity of the rotavirus phosphoprotein NSP5 includes affinity for double-stranded RNA. J Virol, 2002. 76(10): p. 5291-9.
6.Jayaram, H., et al., Rotavirus protein involved in genome replication and packaging exhibits a HIT-like fold. Nature, 2002. 417(6886): p. 311-5.
7.Taraporewala, Z.F., et al., Analysis of a temperature-sensitive mutant rotavirus indicates that NSP2 octamers are the functional form of the protein. J Virol, 2002. 76(14): p. 7082-93.
8.Mirazimi, A., K.E. Magnusson, and L. Svensson, A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum. J Gen Virol, 2003. 84(Pt 4): p. 875-83.
9.Estes, M.K., et al., Synthesis and immunogenicity of the rotavirus major capsid antigen using a baculovirus expression system. J Virol, 1987. 61(5): p. 1488-94.
10.Roseto, A., et al., Structure of rotaviruses as studied by the freeze-drying technique. Virology, 1979. 98(2): p. 471-5.
11.Estes, M.K. and J. Cohen, Rotavirus gene structure and function. Microbiol Rev, 1989. 53(4): p. 410-49.
12.Liu, M., P.A. Offit, and M.K. Estes, Identification of the simian rotavirus SA11 genome segment 3 product. Virology, 1988. 163(1): p. 26-32.
13.Estes, M.K., et al., Cloning and nucleotide sequence of the simian rotavirus gene 6 that codes for the major inner capsid protein. Nucleic Acids Res, 1984. 12(4): p. 1875-87.
14.Glass, R.I., et al., Rotavirus vaccines at the threshold. Nat Med, 1997. 3(12): p. 1324-5.
15.Parashar, U.D., et al., Global illness and deaths caused by rotavirus disease in children. Emerg Infect Dis, 2003. 9(5): p. 565-72.
16.Kapikian, A.Z., et al., Oral administration of human rotavirus to volunteers: induction of illness and correlates of resistance. J Infect Dis, 1983. 147(1): p. 95-106.
17.Isolauri, E., T. Jalonen, and M. Maki, Acute gastroenteritis. Changing pattern of clinical features and management. Acta Paediatr Scand, 1989. 78(5): p. 685-91.
18.Gouvea, V., et al., Detection of group B and C rotaviruses by polymerase chain reaction. J Clin Microbiol, 1991. 29(3): p. 519-23.
19.Saif, L.J. and B. Jiang, Nongroup A rotaviruses of humans and animals. Curr Top Microbiol Immunol, 1994. 185: p. 339-71.
20.Rahman, M., et al., Detection and characterization of human group C rotaviruses in Bangladesh. J Clin Microbiol, 2005. 43(9): p. 4460-5.
21.Hoshino, Y. and A.Z. Kapikian, Classification of rotavirus VP4 and VP7 serotypes. Arch Virol Suppl, 1996. 12: p. 99-111.
22.Phan, T.G., et al., Detection and genetic characterization of group A rotavirus strains circulating among children with acute gastroenteritis in Japan. J Virol, 2007. 81(9): p. 4645-53.
23.Zarate, S., et al., The VP5 domain of VP4 can mediate attachment of rotaviruses to cells. J Virol, 2000. 74(2): p. 593-9.
24.Mebus, C.A., M.B. Rhodes, and N.R. Underdahl, Neonatal calf diarrhea caused by a virus that induces villous epithelial cell syncytia. Am J Vet Res, 1978. 39(7): p. 1223-8.
25.Theil, K.W., et al., Porcine rotavirus-like virus (group B rotavirus): characterization and pathogenicity for gnotobiotic pigs. J Clin Microbiol, 1985. 21(3): p. 340-5.
26.Tsunemitsu, H., et al., Molecular characterization of the major capsid protein VP6 of bovine group B rotavirus and its use in seroepidemiology. J Gen Virol, 2005. 86(Pt 9): p. 2569-75.
27.Chen, C.M., et al., Chinese adult rotavirus is a group B rotavirus. Lancet, 1985. 2(8464): p. 1123-4.
28.Hung, T., et al., Seroepidemiology and molecular epidemiology of the Chinese rotavirus. Ciba Found Symp, 1987. 128: p. 49-62.
29.Hung, T., et al., Waterborne outbreak of rotavirus diarrhoea in adults in China caused by a novel rotavirus. Lancet, 1984. 1(8387): p. 1139-42.
30.Hung, T., et al., Rotavirus-like agent in adult non-bacterial diarrhoea in China. Lancet, 1983. 2(8358): p. 1078-9.
31.Mackow, E.R., et al., Cloning sequencing and expression of the gene encoding the VP2 protein of the human group B rotavirus, ADRV. Virology, 1994. 201(1): p. 162-8.
32.Tao, H., Rotavirus and adult diarrhea. Adv Virus Res, 1988. 35: p. 193-218.
33.Yang, H., S. Chen, and S. Ji, [A novel rotavirus causing large scale of adult diarrhea in Shi Jiazhuang]. Zhonghua Liu Xing Bing Xue Za Zhi, 1998. 19(6): p. 336-8.
34.Hung, T., et al., Seroepidemiology of adult rotavirus. Lancet, 1985. 2(8450): p. 325-6.
35.Penaranda, M.E., et al., Seroepidemiology of adult diarrhea rotavirus in China, 1977 to 1987. J Clin Microbiol, 1989. 27(10): p. 2180-3.
36.Krishnan, T., et al., Emergence of adult diarrhoea rotavirus in Calcutta, India. Lancet, 1999. 353(9150): p. 380-1.
37.Sanekata, T., et al., Human group B rotavirus infections cause severe diarrhea in children and adults in Bangladesh. J Clin Microbiol, 2003. 41(5): p. 2187-90.
38.Carmona, R.C., et al., Characterization of rotavirus strains from hospitalized and outpatient children with acute diarrhoea in Sao Paulo, Brazil. J Med Virol, 2004. 74(1): p. 166-72.
39.Nakata, S., et al., Detection of antibody to group B adult diarrhea rotaviruses in humans. J Clin Microbiol, 1987. 25(5): p. 812-8.
40.Eiden, J.J., et al., Serum antibody response to recombinant major inner capsid protein following human infection with group B rotavirus. J Clin Microbiol, 1994. 32(6): p. 1599-603.
41.Chasey, D., J.C. Bridger, and M.A. McCrae, A new type of atypical rotavirus in pigs. Arch Virol, 1986. 89(1-4): p. 235-43.
42.Otto, P., P. Schulze, and W. Herbst, Demonstration of group C rotaviruses in fecal samples of diarrheic dogs in Germany. Arch Virol, 1999. 144(12): p. 2467-73.
43.Nicolas, J.C., et al., Isolation of a human pararotavirus. Virology, 1983. 124(1): p. 181-4.
44.Espejo, R.T., et al., Characterization of a human pararotavirus. Infect Immun, 1984. 44(1): p. 112-6.
45.Pereira, H.G., et al., An atypical rotavirus detected in a child with gastroenteritis in Rio de Janeiro, Brazil. Mem Inst Oswaldo Cruz, 1983. 78(3): p. 245-50.
46.Bridger, J.C., S. Pedley, and M.A. McCrae, Group C rotaviruses in humans. J Clin Microbiol, 1986. 23(4): p. 760-3.
47.Oishi, I., K. Yamazaki, and Y. Minekawa, An occurrence of diarrheal cases associated with group C rotavirus in adults. Microbiol Immunol, 1993. 37(6): p. 505-9.
48.Iturriza-Gomara, M., et al., Seroepidemiology of group C rotavirus infection in England and Wales. Eur J Epidemiol, 2004. 19(6): p. 589-95.
49.James, V.L., et al., Seroepidemiology of human group C rotavirus in the UK. J Med Virol, 1997. 52(1): p. 86-91.
50.Caul, E.O., et al., Group C rotavirus associated with fatal enteritis in a family outbreak. J Med Virol, 1990. 30(3): p. 201-5.
51.Riepenhoff-Talty, M., et al., Epidemiology of group C rotavirus infection in Western New York women of childbearing age. J Clin Microbiol, 1997. 35(2): p. 486-8.
52.Ojeh, C.K., et al., Development of a biotin-streptavidin-enhanced enzyme-linked immunosorbent assay which uses monoclonal antibodies for detection of group C rotaviruses. J Clin Microbiol, 1992. 30(7): p. 1667-73.
53.Jiang, B., et al., First detection of group C rotavirus in fecal specimens of children with diarrhea in the United States. J Infect Dis, 1995. 172(1): p. 45-50.
54.Steele, A.D. and V.L. James, Seroepidemiology of human group C rotavirus in South Africa. J Clin Microbiol, 1999. 37(12): p. 4142-4.
55.Kuzuya, M., et al., Seroepidemiology of human group C rotavirus in Japan based on a blocking enzyme-linked immunosorbent assay. Clin Diagn Lab Immunol, 2001. 8(1): p. 161-5.
56.Kuzuya, M., et al., Survey of human group C rotaviruses in Japan during the winter of 1992 to 1993. J Clin Microbiol, 1998. 36(1): p. 6-10.
57.Kuzuya, M., et al., Rapid detection of human group C rotaviruses by reverse passive hemagglutination and latex agglutination tests using monoclonal antibodies. J Clin Microbiol, 1993. 31(5): p. 1308-11.
58.Von Bonsdorf, C.H. and L. Svensson, Human serogroup C rotavirus in Finland. Scand J Infect Dis, 1988. 20(5): p. 475-8.
59.Adah, M.I., et al., Detection of human group C rotaviruses in Nigeria and sequence analysis of their genes encoding VP4, VP6, and VP7 proteins. J Med Virol, 2002. 66(2): p. 269-75.
60.Nilsson, M., G. Sigstam, and L. Svensson, Antibody prevalence and specificity to group C rotavirus in Swedish sera. J Med Virol, 2000. 60(2): p. 210-5.
61.Penaranda, M.E., et al., Group C rotavirus infections in patients with diarrhea in Thailand, Nepal, and England. J Infect Dis, 1989. 160(3): p. 392-7.
62.Rasool, N.B., et al., Identification of a human group C rotavirus in Malaysia. J Med Virol, 1994. 43(3): p. 209-11.
63.Rodger, S.M., R.F. Bishop, and I.H. Holmes, Detection of a rotavirus-like agent associated with diarrhea in an infant. J Clin Microbiol, 1982. 16(4): p. 724-6.
64.Wu, H., et al., Serological and genomic characterization of human rotaviruses detected in China. J Med Virol, 1998. 55(2): p. 168-76.
65.Burns, J.W., et al., Characterization of monoclonal antibodies to human group B rotavirus and their use in an antigen detection enzyme-linked immunosorbent assay. J Clin Microbiol, 1989. 27(2): p. 245-50.
66.Rohrmann, G.F., Polyhedrin structure. J Gen Virol, 1986. 67 ( Pt 8): p. 1499-513.
67.Rohrmann, G.F., Baculovirus structural proteins. J Gen Virol, 1992. 73 ( Pt 4): p. 749-61.
68.Harrap, K.A., The structure of nuclear polyhedrosis viruses. II. The virus particle. Virology, 1972. 50(1): p. 124-32.
69.Miller, L.K., Baculoviruses as gene expression vectors. Annu Rev Microbiol, 1988. 42: p. 177-99.
70.Okano, K., et al., Conserved molecular systems of the Baculoviridae. Virology, 2006. 344(1): p. 77-87.
71.Russell, R.L., C.J. Funk, and G.F. Rohrmann, Association of a baculovirus-encoded protein with the capsid basal region. Virology, 1997. 227(1): p. 142-52.
72.Volkman, L.E. and M.D. Summers, Autographa californica nuclear polyhedrosis virus: comparative infectivity of the occluded, alkali-liberated, and nonoccluded forms. J Invertebr Pathol, 1977. 30(1): p. 102-3.
73.Guarino, L.A. and M.D. Summers, Functional mapping of Autographa california nuclear polyhedrosis virus genes required for late gene expression. J Virol, 1988. 62(2): p. 463-71.
74.Thiem, S.M. and L.K. Miller, Identification, sequence, and transcriptional mapping of the major capsid protein gene of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol, 1989. 63(5): p. 2008-18.
75.Carstens, E.B., Mapping the Mutation Site of an Autographa californica Nuclear Polyhedrosis Virus Polyhedron Morphology Mutant. J Virol, 1982. 43(3): p. 809-818.
76.Russell, R.L., M.N. Pearson, and G.F. Rohrmann, Immunoelectron microscopic examination of Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus-infected Lymantria dispar cells: time course and localization of major polyhedron-associated proteins. J Gen Virol, 1991. 72 ( Pt 2): p. 275-83.
77.Smith, G.E., M.J. Fraser, and M.D. Summers, Molecular Engineering of the Autographa californica Nuclear Polyhedrosis Virus Genome: Deletion Mutations Within the Polyhedrin Gene. J Virol, 1983. 46(2): p. 584-593.
78.Smith, G.E., M.D. Summers, and M.J. Fraser, Production of human beta interferon in insect cells infected with a baculovirus expression vector. Mol Cell Biol, 1983. 3(12): p. 2156-65.
79.Vlak, J.M., et al., Functional studies on the p10 gene of Autographa californica nuclear polyhedrosis virus using a recombinant expressing a p10-beta-galactosidase fusion gene. J Gen Virol, 1988. 69 ( Pt 4): p. 765-76.
80.Kitts, P.A., M.D. Ayres, and R.D. Possee, Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucleic Acids Res, 1990. 18(19): p. 5667-72.
81.Tessier, D.C., et al., Enhanced secretion from insect cells of a foreign protein fused to the honeybee melittin signal peptide. Gene, 1991. 98(2): p. 177-83.
82.Luckow, V.A. and M.D. Summers, Signals important for high-level expression of foreign genes in Autographa californica nuclear polyhedrosis virus expression vectors. Virology, 1988. 167(1): p. 56-71.
83.Carbonell, L.F. and L.K. Miller, Baculovirus interaction with nontarget organisms: a virus-borne reporter gene is not expressed in two mammalian cell lines. Appl Environ Microbiol, 1987. 53(7): p. 1412-7.
84.Matsumoto, K., et al., An outbreak of gastroenteritis associated with acute rotaviral infection in schoolchildren. J Infect Dis, 1989. 160(4): p. 611-5.