猪病毒性腹泻多重RT-PCR诊断方法的建立和应用及猪轮状病毒的分离
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摘要
在寒冷季节,猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)和猪A群轮状病毒(PGAR)是引起猪腹泻的主要病原,其临床症状、病理变化和流行病学极为相似,在临床和组织病理学上很难区分。本试验建立了能同时检测猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)和猪A群轮状病毒(PGAR)的多重RT-PCR检测方法。根据GenBank登录的PEDV M基因、TGEV N基因、PGAR VP7基因,利用软件Primer 5.0设计合成三对特异性引物。经条件优化,我们建立了一种能够同时检测PEDV, TGEV和PGAR的多重RT-PCR的方法,并将这种方法和常规的RT-PCR进行了比较。特异性试验中,多重RT-PCR检测方法能够检测到35 pg的TGEV-PEDV-PGAR三联苗的混合RNA。在临床检测中,应用该方法检测了华中地区75份腹泻猪粪样,同时利用常规RT-PCR检测方法对该检测方法的敏感性和特异性进行了分析(PEDV、TGEV、PGAR敏感性分别为92%、100%、100%,特异性均为100%)。该方法敏感性高、特异性强,可用于腹泻样品中PEDV、TGEV和PGAR的检测。
利用已建立的PEDV、TGEV、PGAR多重RT-PCR检测方法对2009年1月至2010年2月来自湖北、河南、湖南、江西、广西等地区部分养殖场的197份猪腹泻粪样进行了检测。结果为PEDV感染58份,占29.44%；TGEV感染1份,占0.51%；PGAR感染15份,占7.61%。哺乳仔猪、保育及育肥猪和母猪三个生长阶段进行了统计分析：在PEDV 58份阳性样品中哺乳仔猪占24.14%、保育及育肥猪占55.17%、母猪占20.69%; TGEV仅从哺乳仔猪检出、保育及育肥猪和母猪未检测出；PGAR 15份阳性样品中哺乳仔猪占13.33%、保育及育肥猪占6.67%、母猪占80.00%。检测结果显示：在寒冷季节,在本次收集的腹泻样品中猪流行性腹泻病毒(PEDV)是引起不同生长阶段猪病毒性腹泻最常见的病原,其次是猪A群轮状病毒。
将三份轮状病毒阳性的腹泻猪粪样接种MA104细胞进行了轮状病毒分离,盲传至第三代开始出现明显细胞病变(CPE),将分离毒株依次命名为：TM-a毒株、WH-a毒株和GX-a毒株。对分离病毒进行了常规RT-PCR鉴定；同时依据GenBank发表的猪A群轮状病毒的VP6和VP7基因序列各设计了一对引物,对三株分离毒株的VP6和VP7基因序列进行了克隆及序列分析。结果显示TM-a株VP6基因与中国武汉人源A群R479毒株亲缘关系更为接近(94.4%),而与其VP7基因序列亲缘性最近的是印度人源A群RMC321毒株(95.1%)；WH-a株VP6基因与中国北京人源A群LL3354毒株亲缘关系更为接近(96.0%),而与其VP7基因亲缘性最近的的是印度人源A群RMC321毒株(94.6%)；GX-a株VP6基因与泰国猪源A群CMP74/01毒株的亲缘性最近(94.4%)。这些进一步证实分离的三株病毒为A群猪轮状病毒以及轮状病毒的多样性和复杂性。
In winter, porcine epidemic diarrhea virus(PEDV),porcine transmissble gastroenteritis virus(TGEV) and porcine group A rotavirus(PGAR) are major agents in viral diarrhea of pigs.the three viruses induce similar clinical signs,lesions and epidemiology. To develop a multiplex reverse transcription polymerase chain reaction (mutiplex RT-PCR) for detection of porcine epidemic diarrhea virus (PEDV), porcine transmissble gastroenteritis virus(TGEV) and porcine group A rotavirus(PGAR). Three pairs of primers targeting the M gene, N gene, VP7 gene of PEDV, TGEV, PGAR were designed respectively. Using the three pairs of primes, A multiplex reverse transcription polymerase chain reaction (mutiplex RT-PCR) was developed.And we compared the mutiplex RT-PCR with routine RT-PCR in the field trail.In the laboratory test, we found that the detection limit of multiplex RT-PCR is 35 pg RNA of TGEV-PEDV-PGAR vaccine. And in the field trail, a total of 75 fecal specimens from pigs with diarrhea were collected in the central area of China. The relative sensitivity and specificity of multiplex RT-PCR were evaluated. The results suggesting that this assay is equal in quality to routine RT-PCR assys (sensitivity: 92%,100%,100% for PEDV, TGEV, PGAR respectively; specificity:100% for all 3 viruses). The results indicated that the multiplex RT-PCR with high sensitivity and specificity provided a new and alternative tool for the detection of PEDV, TGEV and PGAR.
A tatal of 197 fecal samples from swine with diarrheal were collected from HuBei, HeNan, HuNan, JiangXi and GuangXi and tested by a mutiplex RT-PCR that can detect PEDV, TGEV and PGAR. Fifty-eight (29.44%) infections were caused by PEDV, one (0.51%) infection was caused by TGEV and fifteen (7.61%) infections were caused by PGAR. In the PEDV and PGAR positive samples, the suckling piglets,the weaning or growing pigs and the sow accounts for 24.14%,55.17%,20.69% and 13.33%,6.67%, 80.00%. TGEV was only detected in the suckling piglets. The results indicated that porcine epidemic diarrhea virus (PEDV) and porcine group A rotavirus(PGAR) are major agents in enteric diseases of pigs.
Three strains of porcine group A rotavirus(PGAR) named as TM-a, WH-a and GX-a were isolated from fecal specimens of pigs with diarrhea by using MA 104 cell, and produced obvious cytopathic effects. And these viruses were identified by routine RT-PCR.Tow pairs of primers were designed according to VP6 and VP7 sequences of porcine group A rotavirus in GenBank, and VP6 and VP7 gene were amplified by RT-PCR and cloned into pMD18-T vectors. Gene sequences analysis showed that VP6 and VP7 gene sequences of TM-a were more closely related to human rotavirus R479 isolated from WuHan and human rotavirus RMC321 isolated from India; VP6 and VP7 gene sequences of WH-a were more closely related to human rotavirus LL3354 isolated from BeiJing and human rotavirus RMC321 isolated from India; VP6 gene sequence of GX-a was more closely related to porcine rotavirus CMP74/01 isolated from Thailand. These indicated that rotavirus was diversity and complexity.
利用已建立的PEDV、TGEV、PGAR多重RT-PCR检测方法对2009年1月至2010年2月来自湖北、河南、湖南、江西、广西等地区部分养殖场的197份猪腹泻粪样进行了检测。结果为PEDV感染58份,占29.44%；TGEV感染1份,占0.51%；PGAR感染15份,占7.61%。哺乳仔猪、保育及育肥猪和母猪三个生长阶段进行了统计分析：在PEDV 58份阳性样品中哺乳仔猪占24.14%、保育及育肥猪占55.17%、母猪占20.69%; TGEV仅从哺乳仔猪检出、保育及育肥猪和母猪未检测出；PGAR 15份阳性样品中哺乳仔猪占13.33%、保育及育肥猪占6.67%、母猪占80.00%。检测结果显示：在寒冷季节,在本次收集的腹泻样品中猪流行性腹泻病毒(PEDV)是引起不同生长阶段猪病毒性腹泻最常见的病原,其次是猪A群轮状病毒。
将三份轮状病毒阳性的腹泻猪粪样接种MA104细胞进行了轮状病毒分离,盲传至第三代开始出现明显细胞病变(CPE),将分离毒株依次命名为：TM-a毒株、WH-a毒株和GX-a毒株。对分离病毒进行了常规RT-PCR鉴定；同时依据GenBank发表的猪A群轮状病毒的VP6和VP7基因序列各设计了一对引物,对三株分离毒株的VP6和VP7基因序列进行了克隆及序列分析。结果显示TM-a株VP6基因与中国武汉人源A群R479毒株亲缘关系更为接近(94.4%),而与其VP7基因序列亲缘性最近的是印度人源A群RMC321毒株(95.1%)；WH-a株VP6基因与中国北京人源A群LL3354毒株亲缘关系更为接近(96.0%),而与其VP7基因亲缘性最近的的是印度人源A群RMC321毒株(94.6%)；GX-a株VP6基因与泰国猪源A群CMP74/01毒株的亲缘性最近(94.4%)。这些进一步证实分离的三株病毒为A群猪轮状病毒以及轮状病毒的多样性和复杂性。
In winter, porcine epidemic diarrhea virus(PEDV),porcine transmissble gastroenteritis virus(TGEV) and porcine group A rotavirus(PGAR) are major agents in viral diarrhea of pigs.the three viruses induce similar clinical signs,lesions and epidemiology. To develop a multiplex reverse transcription polymerase chain reaction (mutiplex RT-PCR) for detection of porcine epidemic diarrhea virus (PEDV), porcine transmissble gastroenteritis virus(TGEV) and porcine group A rotavirus(PGAR). Three pairs of primers targeting the M gene, N gene, VP7 gene of PEDV, TGEV, PGAR were designed respectively. Using the three pairs of primes, A multiplex reverse transcription polymerase chain reaction (mutiplex RT-PCR) was developed.And we compared the mutiplex RT-PCR with routine RT-PCR in the field trail.In the laboratory test, we found that the detection limit of multiplex RT-PCR is 35 pg RNA of TGEV-PEDV-PGAR vaccine. And in the field trail, a total of 75 fecal specimens from pigs with diarrhea were collected in the central area of China. The relative sensitivity and specificity of multiplex RT-PCR were evaluated. The results suggesting that this assay is equal in quality to routine RT-PCR assys (sensitivity: 92%,100%,100% for PEDV, TGEV, PGAR respectively; specificity:100% for all 3 viruses). The results indicated that the multiplex RT-PCR with high sensitivity and specificity provided a new and alternative tool for the detection of PEDV, TGEV and PGAR.
A tatal of 197 fecal samples from swine with diarrheal were collected from HuBei, HeNan, HuNan, JiangXi and GuangXi and tested by a mutiplex RT-PCR that can detect PEDV, TGEV and PGAR. Fifty-eight (29.44%) infections were caused by PEDV, one (0.51%) infection was caused by TGEV and fifteen (7.61%) infections were caused by PGAR. In the PEDV and PGAR positive samples, the suckling piglets,the weaning or growing pigs and the sow accounts for 24.14%,55.17%,20.69% and 13.33%,6.67%, 80.00%. TGEV was only detected in the suckling piglets. The results indicated that porcine epidemic diarrhea virus (PEDV) and porcine group A rotavirus(PGAR) are major agents in enteric diseases of pigs.
Three strains of porcine group A rotavirus(PGAR) named as TM-a, WH-a and GX-a were isolated from fecal specimens of pigs with diarrhea by using MA 104 cell, and produced obvious cytopathic effects. And these viruses were identified by routine RT-PCR.Tow pairs of primers were designed according to VP6 and VP7 sequences of porcine group A rotavirus in GenBank, and VP6 and VP7 gene were amplified by RT-PCR and cloned into pMD18-T vectors. Gene sequences analysis showed that VP6 and VP7 gene sequences of TM-a were more closely related to human rotavirus R479 isolated from WuHan and human rotavirus RMC321 isolated from India; VP6 and VP7 gene sequences of WH-a were more closely related to human rotavirus LL3354 isolated from BeiJing and human rotavirus RMC321 isolated from India; VP6 gene sequence of GX-a was more closely related to porcine rotavirus CMP74/01 isolated from Thailand. These indicated that rotavirus was diversity and complexity.
引文
1.黄军建等.猪传染性胃肠炎病毒国外株与国内株的比较研究[J].中国畜牧兽医年会,1980，2：13-16.
2.倪艳秀,林继煌,何孔旺,还红华.猪流行性腹泻研究概况[J].畜牧与兽医,2001,33(1)：38-39.
3.冉智光,王玉春.猪传染性胃肠炎病毒分子生物学研究新进展[J].天津畜牧兽医,1998,15(4)：4-6
4.田小艳,孙华,邓雨修,李春梅,徐贵娟,宋延华.猪轮状病毒分离与鉴定[J].畜牧与兽医,2009,41(10)：85-87.
5.余丽芸,刘建柱.多重PCR检测猪瘟病毒、猪细小病毒和猪伪狂犬病病毒的研究[J].动物医学进展,2004,25(26)：75-77.
6. Agbalika F, Hartemann P, Foliguet JM. Trypsin-treated Ma-104:a sensitive cell line for isolating enteric viruses from environmental samples[J]. Appl Environ Microbiol, 1984,47(2):378-380.
7. Anton IM, Sune C, Meloen RH, Borras-Cuesta F, Enjuanes L. A transmissible gastroenteritis coronavirus nucleoprotein epitope elicits T helper cells that collaborate in the in vitro antibody synthesis to the three major structural viral proteins[J]. Virology,1995,212(2):746-751.
8. Arriba ML, Carvajal A, Pozo J, Rubio P. Mucosal and systemic isotype-specific antibody responses and protection in conventional pigs exposed to virulent or attenuated porcine epidemic diarrhoea virus [J]. Veterinary Immunology and Immunopathology,2002,85(1-2):85-97
9. Benfield DA, Stotz IJ, Nelson EA, Groon KS. Comparison of a commercial enzyme-linked immunosorbent assay with electron microscopy, fluorescent antibody, and virus isolation for the detection of bovine and porcine rotavirus[J]. Am J Vet Res, 1984,45(10):1998-2002.
10. Britton P, Carmenes RS, Page KW, Garwes DJ, Parra F. Sequence of the nucleoprotein gene from a virulent British field isolate of transmissible gastroenteritis virus and its expression in Saccharomyces cerevisiae[J]. Mol Microbiol,1988,2(1):89-99.
11. Bridgen A, Kocherhans R, Tobler K, Laude H, Ackermann M. Sequence determination of the nucleocapsid protein gene of the porcine epidemic diarrhea virus confirms that this is a coronavirus related to human coronavirus 229E and porcine transmissible gastroenteritis virus[J]. J Gen Virol,1993,74:1795-1804.
12. Bosch BJ, van der Zee R, de Haan CA, Rottier PJ. The coronavirus spike protein is a class I virus fusion protein:structural and functional characterization of the fusion core complex[J]. J Virol,2003,77(16):8801-8811.
13. Broor S, Ghosh D, Mathur P. Molecular epidemiology of rotaviruses in India[J]. Indian J Med Res,2003,118:59-67.
14. Banyai K, Esona MD, Kerin TK, Hull JJ, Mijatovic S, Vasconez N, Torres C, de Filippis AM, Foytich KR, Gentsch JR. Molecular characterization of a rare, human-porcine reassortant rotavirus strain G11P[6] from Ecuador[J]. Arch Virol, 2009,154(11):1823-1829.
15. Ben Salem AN, Chupin Sergei A, Bjadovskaya Olga P, Andreeva Olga G, Mahjoub A, Prokhvatilova Larissa B. Multiplex nested RT-PCR for the detection of porcine enteric viruses[J]. J Virol Methods,2010,165(2):283-93.
16. Carvajal A, Lanza I, Diego R, Rubio P, Carmenes P. Seroprevalence of porcine epidemic diarrhea virus infection among different types of breeding swine farms in Spain[J]. Prev Vet Med,1995,23:33-40.
17. Coulson BS. VP4 and VP7 typing using monoclonal antibodies [J]. Arch Virol Suppl, 1996,12:113-118.
18. Cavanagh D. Nidovirales:a new order comprising Coronaviridae and Arteriviridae[J]. Arch Virol,1997,142:629-633.
19. Chae C, Kim O, Choi C, Min K, Cho WS, Kim J, Tai JH. Prevalence of porcine epidemic diarrhea virus and transmissible gastroenteritis virus infection in Korean pigs[J]. Vet Rec,2000,18:606-608.
20. Carman S, Josephson G, McEwen B, Maxie G, Antochi M, Eernisse K, Nayar G, Halbur P, Erickson G, Nilsson E. Field validation of a commercial blocking ELISA to differentiate antibody to transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus and to identify TGEV-infected swine herds [J].J Vet Diagn Invest,2002,14(2):97-105.
21. Chang SH, Bae JL, Kang TJ, Kim J, Chung GH, Lim CW, Laude H, Yang MS, Jang YS. Identification of the epitope region capable of inducing neutralizing antibodies against the porcine epidemic diarrhea virus[J]. Mol Cells,2002,14(2):295-299.
22. Carman S, Josephson G, McEwen B, Maxie G, Antochi M, Eernisse K, Nayar G, Halbur P, Erickson G, Nilsson E. Field validation of a commercial blocking ELISA to differentiate antibody to transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus and to identify TGEV-infected swine herds[J]. J Vet Diagn Invest,2002,14(2):97-105.
23. Chen R, Huang W, Lin Z, Zhou Z, Yu H, Zhu D. Development of a novel real-time RT-PCR assay with LUX primer for the detection of swine transmissible gastroenteritis virus[J]. J Virol Methods,2004,122(1):57-61.
24. Chen JF, Sun DB, Wang CB, Shi HY, Cui XC, Liu SW, Qiu HJ, Feng L. Molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in China[J]. Virus Genes,2008,36(2):355-364.
25. Debouck P, Pensaert MB.Experimental infection of pigs with a new porcine enteric coronavirus CV777[J]. Am J Vet Res,1980,41:219-223
26. Delmas B, Rasschaert D, Godet M, Gelfi J, Laude H. Four major antigenic site of the cornavirus transmissible gastroenteritis virus are locate on the amino-terminal half of spike glycoprotein S[H].J Gen Virol,1990,71:1313-1323.
27. Duarte M, Laude H. Sequence of the spike protein of the porcine epidemic diarrhoea virus[J]. J Gen Virol,1994,75 (5):1195-1200.
28. Duarte M, Tobler K, Bridgen A, Rasschaert D, Ackermann M, Laude H. Sequence analysis of the porcine epidemic diarrhea virus genome between the nucleocapsid and spike protein genes reveals a polymorphic ORF[J]. Virology,1994,198(2): 466-476.
29. Dae-Sub Song, Bo-Kyu Kang, Sang-Sun Lee, Jeong-Sun Yang, Hyoung-Joon Moon, Jin-Sik Oh, Gun-Woo Ha, Yong-Suk Jang, Bong-Kyun Park. Use of an internal control in a quantitative RT-PCR assay for quantitation of porcine epidemic diarrhea virus shedding in pigs[J]. Journal of Virological Methods,2006,133(1):27-33
30. De Grazia S, Giammanco GM, Potgieter CA, Matthijnssens J, Banyai K, Platia MA, Colomba C, Martella V. Unusual assortment of segments in 2 rare human rotavirus genomes[J]. Emerg Infect Dis,2010,16(5):859-862.
31. Enjuanes L, Smerdou C, Castilla J, Anton IM, Torres JM, Sola I, Golvano J, Sanchez JM, Pintado B. Development of protection against coronavirus induced diseases [J]. Adv Exp Med Biol,1995,380:197-211
32. Elschner M, Prudlo J, Hotzel H, Otto P, Sachse K. Nested reverse transcriptase-polymerase chain reaction for the detection of group A rotaviruses[J]. J Vet Med B Infect Dis Vet Public Health,2002,49(2):77-81.
33. Enrica Sozzi, Andrea Luppi, Davide Lelli, Ana Moreno Martin, Elena Canelli, Emiliana Brocchi, Antonio Lavazza, Paolo Cordioli. Comparison of enzyme-linked immunosorbent assay and RT-PCR for the detection of porcine epidemic diarrhoea virus[J]. Research in Veterinary Science,2010,31(1):166-168
34. Greenberg H, McAuliffe V, Valdesuso J, Wyatt R, Flores J, Kalica A, Hoshino Y, Singh N. Serological analysis of the subgroup protein of rotavirus using monoclonal antibodies[J]. Infect Immun,1983,39(1):91-99.
35. Gebauer F, Posthumus WP, Correa I, Sune C, Smerdou C, Sanchez CM, Lenstra JA, Meloen RH, Enjuanes L. Residues Involved in the antigenic sites of transmissible gastroenteritis coronavirus S Glycoprotein[J]. Virology,1991,18(3):225-238
36. Godet M, Grosclaude J, Delmas B, Laude H. Major receptor-binding and neutralization determinants are located within the same domain of the transmissible gastroenteritis virus (coronavirus) spike protein[J]. J Virol,1994,68(12):8008-8016.
37. Harada K, Kaji T, Kumagai T, Sasahara J. Studies on transmissible gastroenteritis in pigs. Ⅳ. Physicochemical and biological properties of TGE virus[J]. J Vet Sci,1968, 8(3):140-147.
38. Hofmann M, Wyler R. Propagation of the virus of porcine epidemic diarrhea in cell culture[J]. J Clin Microbiol,1988,26:2235-2239.
39. Hoshino Y,Honma S,Jones RW,Ross J,Santos N,Gentsch JR,Kapikian AZ,Hesse RA. A porcine G9 rotavirus strain shares neutralization and VP7 phylogenetic sequence lineage 3 characteristics with contemporary human G9 rotavirus strains[J]. Virology, 2005,5332(1):177-188.
40. Jackwood DJ, Kwon HM, Saif LJ. Molecular differentiation of transmissible gastroenteritis virus and porcine respiratory coronavirus strains.Correlation with antigenicity and pathogenicity[J]. Adv Exp Med Biol,1995,380:35-41.
41. Jackwood MW, Hilt DA, Callison SA, Lee CW, Plaza H, Wade E. Spike glycoprotein cleavage recognition site analysis of infectious bronchitis virus[J]. Avian Dis,2001, 45(2):366-372.
42. Jain V, Das BK, Bhan MK, Glass RI, Gentsch JR; Indian Strain Surveillance Collaborating Laboratories. Great diversity of group A rotavirus strains and high prevalence of mixed rotavirus infections in India[J]. J Clin Microbiol,2001,39(10): 3524-3529.
43. Jung K, Kang BK, Lee CS, Song DS. Impact of porcine group A rotavirus co-infection on porcine epidemic diarrhea virus pathogenicity in piglets[J]. Res Vet Sci,2008,84(3):502-506.
44. Kemeny LJ, Woods RD. Quantitative transmissible gastroenteritis virus shedding patterns in lactating sows[J]. Am J Vet Res,1977,38(3):307-310.
45. Knuchel M, Ackermann M., Muller HK., Kihm U. An ELISA for detection of antibodies against porcine epidemic diarrhoea virus (PEDV) based on the specific solubility of the viral surface glycoprotein[J]. Veterinary Microbiology,1992,32(2): 117-134.
46. Kim L, Hayes J, Lewis P, Parwani AV, Chang KO, Saif LJ. Molecular characterization and pathogenesis of transmissible gastroenteritis virus(TGEV) and porcine respiratory coronavirus(PRCV) field isolates cocirculating in a swine herd[J]. Arch Virol,2000,145(6):1133-1147.
47. Kim L, Hayes J, Lewis P, Parwani AV, Chang KO, Saif LJ. Molecular characterization and pathogenesis of transmissible gastroenteritis coronavirus (TGEV) and porcine respiratory coronavirus (PRCV) field isolates co-circulating in a swine herd[J]. Arch Virol,2000,145(6):1133-1147.
48. Kim B, Chae C. In situ hybridization for the detection of transmissible gastroenteritis virus in pigs and comparison with other methods[J]. Can J Vet Res,2001,65(1): 33-37.
49. Kim SY, Song DS, Park BK. Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT-PCR[J]. J Vet Diagn Invest, 2001,13:516-520.
50. Kocherhans R, Bridgen A, Ackermann M, Tobler K. Completion of the porcine epidemic diarrhoea coronavirus (PEDV) genome sequence[J]. Virus Genes,2001, 23(2):137-144.
51. Kim SY, Song DS, Park BK..Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT-PCR[J]. J Vet Diagn Invest, 2001,13:516-520.
52. Kim O, Chae C. Experimental Infection of Piglets with a Korean Strain of Porcine Epidemic Diarrhoea Virus[J]. Journal of Comparative Pathology,2003,129(1): 55-60.
53. Kwonil Jung, Junghyum Kim, Okjin Kim, Bongtae Kim, Chanhee Chae. Differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in formalin-fixed paraffin-embedded tissues by multiplex RT-nested PCR and comparison with in situ hybridization[J]. Journal of Virological Methods,2003,108(1):41-47
54. Kwonil Jung, Chanhee Chae. RT-PCR-based dot blot hybridization for the detection and differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in fecal samples using a non-radioactive digoxigenin cDNA probe[J]. Journal of Virological Methods,2005,123(2):141-146.
55. Kim SH, Kim IJ, Pyo HM, Tark DS, Song JY, Hyun BH. Multiplex real-time RT-PCR for the simultaneous detection and quantification of transmissible gastroenteritis virus and porcine epidemic diarrhea virus[J]. J Virol Methods,2007,146(1-2): 172-177.
56. Kang JH, Kwon DH, Chung TW, Kim YD, Lee HG, Kim JW, Choe IS, Kim KW, Lim JS, Song EY, Kim CH. Development of a simple and rapid immunochromatographic strip test for diarrhea-causative porcine rotavirus in swine stool[J]. J Virol Methods, 2007,146(1-2):74-79.
57. Kim HJ, Park SI, Ha TP, Jeong YJ, Kim HH, Kwon HJ, Kang MI, Cho KO, Park SJ. Detection and genotyping of Korean porcine rotaviruses[J]. Vet Microbiol,2010.
58. Lanza I, Rubio P, Munoz M, Carmenes P. Comparison of a monoclonal antibody capture ELISA (MACELISA) to indirect ELISA and virus neutralization test for the serodiagnosis of transmissible gastroenteritis virus[J]. J Vet Diagn Invest,1993,5(1): 21-25.
59. Liu C, Kokuho T, Kubota T, Watanabe S, Inumaru S,Yokomizo Y, Onodera T:A serodiagnostic ELISA using recombinant antigen of swine transmissible gastroenteritis virus nucleoprotein[J]. J Vet Med Sci,2001,63(11):1253-1256.
60. Li D, Gu AZ, Yang W, He M, Hu XH, Shi HC. An integrated cell culture and reverse transcription quantitative PCR assay for detection of infectious rotaviruses in environmental waters[J]. J Microbiol Methods,2010.
61. Lee DK, Park CK, Kim SH, Lee C. Heterogeneity in spike protein genes of porcine epidemic diarrhea viruses isolated in Korea[J]. Virus Res,2010,149(2):175-182.
62. Martella V, Ciarlet M, Banyai K, Lorusso E, Cavalli A, Corrente M, Elia G, Arista S, Camero M, Desario C, Decaro N, Lavazza A, Buonavoglia C. Identification of a novel VP4 genotype carried by a serotype G5 porcine rotavirus strain[J]. Virology, 2006,346(2):301-311.
63. Min BS, Noh YJ, Shin JH, Baek SY, Min KI, Ryu SR, Kim BG, Park MK, Choi SE, Yang EH, Park SN, Hur SJ, Ahn BY. Assessment of the quantitative real-time polymerase chain reaction using a cDNA standard for human group A rotavirus[J]. J Virol Methods,2006,137(2):280-286.
64. Martelli P, Lavazza A, Nigrelli AD, Merialdi G, Alborali LG, Pensaert MB. Epidemic of diarrhoea caused by porcine epidemic diarrhoea virus in Italy[J]. Vet Rec,2008, 162(10):307-310.
65. Martella V, Colombrita D, Lorusso E, Draghin E, Fiorentini S, De Grazia S, Banyai K, Ciarlet M, Caruso A, Buonavoglia C. Detection of a porcine-like rotavirus in a child with enteritis in Italy[J]. J Clin Microbiol,2008,46(10):3501-3507.
66. Matthijnssens J, Ciarlet M, Heiman E, Arijs I, Delbeke T, McDonald SM, Palombo EA, Iturriza-Gomara M, Maes P, Patton JT, Rahman M,Van Ranst M. Full genome-based classification of rotaviruses reveals a common origin between human Wa-Like and porcine rotavirus strains and human DS-1-like and bovine rotavirus strains[J]. J Virol,2008,82(7):3204-3219.
67. Mulherin E, Bryan J, Beltman M, OGrady L, Pidgeon E, Garon L, Lloyd A, Bainbridge J, O'Shea H, Whyte P, Fanning S. Molecular characterisation of a bovine-like rotavirus detected from a giraffe [J]. BMC Vet Res,2008,4:46.
68. Mukherjee A, Dutta D, Ghosh S, Bagchi P, Chattopadhyay S, Nagashima S, Kobayashi N, Dutta P, Krishnan T, Naik TN, Chawla-Sarkar M. Full genomic analysis of a human group A rotavirus G9P[6] strain from Eastern India provides evidence for porcine-to-human interspecies transmission[J]. Arch Virol,2009,154(5): 733-746.
69. Miyazaki A, Fukuda M, Kuga K, Takagi M, Tsunemitsu H. Prevalence of Antibodies against Transmissible Gastroenteritis Virus and Porcine Respiratory Coronavirus among Pigs in Six Regions in Japan[J]. J Vet Med Sci,2010.
70. Nakagomi O, Nakagomi T, Akatani K, Ikegami N. Identification of rotavirus genogroups by RNA-RNA hybridization[J]. Mol Cell Probes,1989,3(3):251-261.
71. Nakagomi O, Nakagomi T. Molecular evidence for naturally occurring single VP7 gene substitution reassortant between human rotaviruses belonging to two different genogroups[J]. Arch Virol,1991,119(1-2):67-81.
72. Nakagomi O,Nakagomi T. Genomic relationships among rotaviruses recovered from various animal species as revealed by RNA-RNA hybridization assays[J]. Res Vet Sci, 2002,73(3):207-214.
73. Oh JS, Song DS, Yang JS, Song JY, Moon HJ, Kim TY, Park BK. Comparison of an enzyme-linked immunosorbent assay with serum neutralization test for serodiagnosis of porcine epidemic diarrhea virus infection[J]. J Vet Sci,2005,6(4):349-52.
74. Pritchard GC, Panton DJ, Wibberley G, Ibata G. Transmissible gastroenteritis and porcine epidemic diarrhea in Britain[J]. Vet Rec,1999,144:616-618.
75. Paton D, Ibata G, Sands J, McGoldrick A. Detection of transmissible gastroenteritis virus by RT-PCR and differentiation from porcine respiratory coronavirus[J]. J Virol Methods,1997,66(2):303-309.
76. Pritchard GC, Paton DJ, Wibberley G, Ibata G.transmissible gastroenteritis and porcine epidemic diarrhoea in Britain[J]. J Vet Res,1999,144:616-618.
77. Prabha S, Verghese S. Detection of porcine rotavirus from tissue and faecal specimens[J]. Indian J Med Microbiol,2009,27(2):149-152.
78. Park JE, Cruz DJ, Shin HJ. Trypsin-induced hemagglutination activity of porcine epidemic diarrhea virus[J]. Arch Virol,2010,155(4):595-599.
79. Riffault S, Grosclaude J, Vayssier M, Laude H, Charley B. Reconstituted coronavirus TGEV virosomes lose the virus ability to induce porcine interferon-alpha production[J]. Vet Res,1997,28(1):77-86
80. Rodak L, Smid B, Nevorankova Z, Valicek L, Smitalova R. Use of monoclonal antibodies in blocking ELISA detection of transmissible gastroenteritis virus in faeces of piglets [J].J Vet Med B Infect Dis Vet Public Health,2005,52(3):105-11.
81. Rodak L, Smitalova R, Smid B, Nevorankova Z, Valicek L, Psikal I. Comparing electron microscopy and a competitive blocking ELISA in the detection of rotaviruses in porcine faeces[J]. Vet J,2009.
82. Saif LJ, Terrett LA, Miller KL, Cross RF. Serial propagation of porcine group C rotavirus (pararotavirus) in a continuous cell line and characterization of the passaged virus[J]. J Clin Microbiol,1988,26:1277-1282.
83. Sirinarumitr T, Siddle S, Grahare F. Porcine transmissible gastroenteritis virus induce apoptosis inswine testes cell culture[J]. Archives of Virology,1998,143(12): 2471-2485.
84. Sestak K, Zhou Z, Shoup DI, Saif LJ. Evaluation of the baculovirus-expressed S glycoprotein of transmissible gastroenteritis virus(TGEV) as antigen in a competition ELISA to differentiate porcine respiratory coronavirus from TGEV antiboies in pig[J]. J Vet Diagn Invest,1999,11(3):205-14.
85. Santos N, Lima RC, Nozawa CM, Linhares RE, Gouvea V. Detection of porcine rotavirus type G9 and of a mixture of types G1 and G5 associated with Wa-like VP4 specificity:evidence for natural human-porcine genetic reassortment[J]. J Clin Microbiol,1999,37(8):2734-2736.
86. Sestak K, Zhou Z, Shoup DI, Saif LJ. Evaluation of the baculovirus-expressed S glycoprotein of transmissible gastroenteritis virus (TGEV) as antigen in a competition ELISA to differentiate porcine respiratory coronavirus from TGEV antibodies in pigs[J]. J Vet Diagn Invest,1999,11(3):205-214.
87. Song DS, Kang BK, Oh JS, Ha GW, Yang JS, Moon HJ, Jang YS, Park BK. Multiplex reverse transcription-PCR for rapid differential detection of porcine epidemic diarrhea virus,transmissible gastroenteritis,and porcine group A rotavirus[J]. J Vet Diagn Invest,2006,18:278-281.
88. Sasirekha Ramani, Miren Iturriza-Gomara, Atanu Kumar Jana, Kurien Anil Kuruvilla, James J. Gray, David W. Brown, Gagandeep Kang. Whole genome characterization of reassortant G10P[11] strain (N155) from a neonate with symptomatic rotavirus infection:Identification of genes of human and animal rotavirus origin[J]. Journal of Clinical Virology,2009,45(3):237-244.
89. Sergeev OV. Porcine epidemic diarrhea[J]. Vopr Virusol,2009,54(2):4-8.
90. Theil KW, Saif LJ, Bohl EH, Agnes AG, Kohler EM. Concurrent porcine rotaviral and transmissible gastroenteritis viral infections in a three-day-old conventional pig[J]. Am J Vet Res,1979,40(5):719-721.
91. Vemulapalli R, Gulani J, Santrich C. A real-time TaqMan RT-PCR assay with an internal amplification control for rapid detection of transmissible gastroenteritis virus in swine fecal samples[J]. J Virol Methods,2009,162(1-2):231-235.
92. Woode GN, Bridger J, Hall GA, Jones JM, Jackson G The isolation of reovirus-like agents (rota-viruses) from acute gastroenteritis of piglets[J]. J Med Microbiol,1976, 9:203-209.
93. Wyatt RG, James HD Jr, Pittman AL, Hoshino Y, Greenberg HB, Kalica AR, Flores J, Kapikian AZ. Direct isolation in cell culture of human rotaviruses and their characterization into four serotypes[J]. J Clin Microbiol,1983,18(2):310-7.
94. Ward RL, Nakagomi O, Knowlton DR, McNeal MM, Nakagomi T, Clemens JD, Sack DA, Schiff GM. Evidence for natural reassortants of human rotaviruses belonging to different genogroups[J]. J Virol,1990,64(7):3219-3225
95. Wang C, Chen J, Shi H, Qiu HJ, Xue F, Liu S, Liu C, Zhu Y, Almazan F, Enjuanes L, Feng L. Rapid differentiation of vaccine strain and Chinese field strains of transmissible gastroenteritis virus by restriction fragment length polymorphism of the N gene[J]. Virus Gene,2010,10(4):81-88
96. Wang C, Chen J, Shi H, Qiu H, Xue F, Liu C, Zhu Y, Liu S, Almazan F, Enjuanes L, Feng L. Molecular characterization of a Chinese vaccine strain of transmissible gastroenteritis virus:mutations that may contribute to attenuation[J]. Virus Gene, 2010,40(3):4030-409.
97. Wang YH, Kobayashi N, Nagashima S, Zhou X, Ghosh S, Peng JS, Hu Q, Zhou DJ, Yang ZQ. Full genomic analysis of a porcine-bovine reassortant G4P[6] rotavirus strain R479 isolated from an infant in China[J]. J Med Virol,2010,82(6):1094-1102.
98. Xi-Lin Hou, Li-Yun Yu, Jianzhu Liu. Development and evaluation of enzyme-linked immunosorbent assay based on recombinant nucleocapsid protein for detection of porcine epidemic diarrhea (PEDV) antibodies[J]. Veterinary Microbiology,2007, 123(1-3):86-92
2.倪艳秀,林继煌,何孔旺,还红华.猪流行性腹泻研究概况[J].畜牧与兽医,2001,33(1)：38-39.
3.冉智光,王玉春.猪传染性胃肠炎病毒分子生物学研究新进展[J].天津畜牧兽医,1998,15(4)：4-6
4.田小艳,孙华,邓雨修,李春梅,徐贵娟,宋延华.猪轮状病毒分离与鉴定[J].畜牧与兽医,2009,41(10)：85-87.
5.余丽芸,刘建柱.多重PCR检测猪瘟病毒、猪细小病毒和猪伪狂犬病病毒的研究[J].动物医学进展,2004,25(26)：75-77.
6. Agbalika F, Hartemann P, Foliguet JM. Trypsin-treated Ma-104:a sensitive cell line for isolating enteric viruses from environmental samples[J]. Appl Environ Microbiol, 1984,47(2):378-380.
7. Anton IM, Sune C, Meloen RH, Borras-Cuesta F, Enjuanes L. A transmissible gastroenteritis coronavirus nucleoprotein epitope elicits T helper cells that collaborate in the in vitro antibody synthesis to the three major structural viral proteins[J]. Virology,1995,212(2):746-751.
8. Arriba ML, Carvajal A, Pozo J, Rubio P. Mucosal and systemic isotype-specific antibody responses and protection in conventional pigs exposed to virulent or attenuated porcine epidemic diarrhoea virus [J]. Veterinary Immunology and Immunopathology,2002,85(1-2):85-97
9. Benfield DA, Stotz IJ, Nelson EA, Groon KS. Comparison of a commercial enzyme-linked immunosorbent assay with electron microscopy, fluorescent antibody, and virus isolation for the detection of bovine and porcine rotavirus[J]. Am J Vet Res, 1984,45(10):1998-2002.
10. Britton P, Carmenes RS, Page KW, Garwes DJ, Parra F. Sequence of the nucleoprotein gene from a virulent British field isolate of transmissible gastroenteritis virus and its expression in Saccharomyces cerevisiae[J]. Mol Microbiol,1988,2(1):89-99.
11. Bridgen A, Kocherhans R, Tobler K, Laude H, Ackermann M. Sequence determination of the nucleocapsid protein gene of the porcine epidemic diarrhea virus confirms that this is a coronavirus related to human coronavirus 229E and porcine transmissible gastroenteritis virus[J]. J Gen Virol,1993,74:1795-1804.
12. Bosch BJ, van der Zee R, de Haan CA, Rottier PJ. The coronavirus spike protein is a class I virus fusion protein:structural and functional characterization of the fusion core complex[J]. J Virol,2003,77(16):8801-8811.
13. Broor S, Ghosh D, Mathur P. Molecular epidemiology of rotaviruses in India[J]. Indian J Med Res,2003,118:59-67.
14. Banyai K, Esona MD, Kerin TK, Hull JJ, Mijatovic S, Vasconez N, Torres C, de Filippis AM, Foytich KR, Gentsch JR. Molecular characterization of a rare, human-porcine reassortant rotavirus strain G11P[6] from Ecuador[J]. Arch Virol, 2009,154(11):1823-1829.
15. Ben Salem AN, Chupin Sergei A, Bjadovskaya Olga P, Andreeva Olga G, Mahjoub A, Prokhvatilova Larissa B. Multiplex nested RT-PCR for the detection of porcine enteric viruses[J]. J Virol Methods,2010,165(2):283-93.
16. Carvajal A, Lanza I, Diego R, Rubio P, Carmenes P. Seroprevalence of porcine epidemic diarrhea virus infection among different types of breeding swine farms in Spain[J]. Prev Vet Med,1995,23:33-40.
17. Coulson BS. VP4 and VP7 typing using monoclonal antibodies [J]. Arch Virol Suppl, 1996,12:113-118.
18. Cavanagh D. Nidovirales:a new order comprising Coronaviridae and Arteriviridae[J]. Arch Virol,1997,142:629-633.
19. Chae C, Kim O, Choi C, Min K, Cho WS, Kim J, Tai JH. Prevalence of porcine epidemic diarrhea virus and transmissible gastroenteritis virus infection in Korean pigs[J]. Vet Rec,2000,18:606-608.
20. Carman S, Josephson G, McEwen B, Maxie G, Antochi M, Eernisse K, Nayar G, Halbur P, Erickson G, Nilsson E. Field validation of a commercial blocking ELISA to differentiate antibody to transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus and to identify TGEV-infected swine herds [J].J Vet Diagn Invest,2002,14(2):97-105.
21. Chang SH, Bae JL, Kang TJ, Kim J, Chung GH, Lim CW, Laude H, Yang MS, Jang YS. Identification of the epitope region capable of inducing neutralizing antibodies against the porcine epidemic diarrhea virus[J]. Mol Cells,2002,14(2):295-299.
22. Carman S, Josephson G, McEwen B, Maxie G, Antochi M, Eernisse K, Nayar G, Halbur P, Erickson G, Nilsson E. Field validation of a commercial blocking ELISA to differentiate antibody to transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus and to identify TGEV-infected swine herds[J]. J Vet Diagn Invest,2002,14(2):97-105.
23. Chen R, Huang W, Lin Z, Zhou Z, Yu H, Zhu D. Development of a novel real-time RT-PCR assay with LUX primer for the detection of swine transmissible gastroenteritis virus[J]. J Virol Methods,2004,122(1):57-61.
24. Chen JF, Sun DB, Wang CB, Shi HY, Cui XC, Liu SW, Qiu HJ, Feng L. Molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in China[J]. Virus Genes,2008,36(2):355-364.
25. Debouck P, Pensaert MB.Experimental infection of pigs with a new porcine enteric coronavirus CV777[J]. Am J Vet Res,1980,41:219-223
26. Delmas B, Rasschaert D, Godet M, Gelfi J, Laude H. Four major antigenic site of the cornavirus transmissible gastroenteritis virus are locate on the amino-terminal half of spike glycoprotein S[H].J Gen Virol,1990,71:1313-1323.
27. Duarte M, Laude H. Sequence of the spike protein of the porcine epidemic diarrhoea virus[J]. J Gen Virol,1994,75 (5):1195-1200.
28. Duarte M, Tobler K, Bridgen A, Rasschaert D, Ackermann M, Laude H. Sequence analysis of the porcine epidemic diarrhea virus genome between the nucleocapsid and spike protein genes reveals a polymorphic ORF[J]. Virology,1994,198(2): 466-476.
29. Dae-Sub Song, Bo-Kyu Kang, Sang-Sun Lee, Jeong-Sun Yang, Hyoung-Joon Moon, Jin-Sik Oh, Gun-Woo Ha, Yong-Suk Jang, Bong-Kyun Park. Use of an internal control in a quantitative RT-PCR assay for quantitation of porcine epidemic diarrhea virus shedding in pigs[J]. Journal of Virological Methods,2006,133(1):27-33
30. De Grazia S, Giammanco GM, Potgieter CA, Matthijnssens J, Banyai K, Platia MA, Colomba C, Martella V. Unusual assortment of segments in 2 rare human rotavirus genomes[J]. Emerg Infect Dis,2010,16(5):859-862.
31. Enjuanes L, Smerdou C, Castilla J, Anton IM, Torres JM, Sola I, Golvano J, Sanchez JM, Pintado B. Development of protection against coronavirus induced diseases [J]. Adv Exp Med Biol,1995,380:197-211
32. Elschner M, Prudlo J, Hotzel H, Otto P, Sachse K. Nested reverse transcriptase-polymerase chain reaction for the detection of group A rotaviruses[J]. J Vet Med B Infect Dis Vet Public Health,2002,49(2):77-81.
33. Enrica Sozzi, Andrea Luppi, Davide Lelli, Ana Moreno Martin, Elena Canelli, Emiliana Brocchi, Antonio Lavazza, Paolo Cordioli. Comparison of enzyme-linked immunosorbent assay and RT-PCR for the detection of porcine epidemic diarrhoea virus[J]. Research in Veterinary Science,2010,31(1):166-168
34. Greenberg H, McAuliffe V, Valdesuso J, Wyatt R, Flores J, Kalica A, Hoshino Y, Singh N. Serological analysis of the subgroup protein of rotavirus using monoclonal antibodies[J]. Infect Immun,1983,39(1):91-99.
35. Gebauer F, Posthumus WP, Correa I, Sune C, Smerdou C, Sanchez CM, Lenstra JA, Meloen RH, Enjuanes L. Residues Involved in the antigenic sites of transmissible gastroenteritis coronavirus S Glycoprotein[J]. Virology,1991,18(3):225-238
36. Godet M, Grosclaude J, Delmas B, Laude H. Major receptor-binding and neutralization determinants are located within the same domain of the transmissible gastroenteritis virus (coronavirus) spike protein[J]. J Virol,1994,68(12):8008-8016.
37. Harada K, Kaji T, Kumagai T, Sasahara J. Studies on transmissible gastroenteritis in pigs. Ⅳ. Physicochemical and biological properties of TGE virus[J]. J Vet Sci,1968, 8(3):140-147.
38. Hofmann M, Wyler R. Propagation of the virus of porcine epidemic diarrhea in cell culture[J]. J Clin Microbiol,1988,26:2235-2239.
39. Hoshino Y,Honma S,Jones RW,Ross J,Santos N,Gentsch JR,Kapikian AZ,Hesse RA. A porcine G9 rotavirus strain shares neutralization and VP7 phylogenetic sequence lineage 3 characteristics with contemporary human G9 rotavirus strains[J]. Virology, 2005,5332(1):177-188.
40. Jackwood DJ, Kwon HM, Saif LJ. Molecular differentiation of transmissible gastroenteritis virus and porcine respiratory coronavirus strains.Correlation with antigenicity and pathogenicity[J]. Adv Exp Med Biol,1995,380:35-41.
41. Jackwood MW, Hilt DA, Callison SA, Lee CW, Plaza H, Wade E. Spike glycoprotein cleavage recognition site analysis of infectious bronchitis virus[J]. Avian Dis,2001, 45(2):366-372.
42. Jain V, Das BK, Bhan MK, Glass RI, Gentsch JR; Indian Strain Surveillance Collaborating Laboratories. Great diversity of group A rotavirus strains and high prevalence of mixed rotavirus infections in India[J]. J Clin Microbiol,2001,39(10): 3524-3529.
43. Jung K, Kang BK, Lee CS, Song DS. Impact of porcine group A rotavirus co-infection on porcine epidemic diarrhea virus pathogenicity in piglets[J]. Res Vet Sci,2008,84(3):502-506.
44. Kemeny LJ, Woods RD. Quantitative transmissible gastroenteritis virus shedding patterns in lactating sows[J]. Am J Vet Res,1977,38(3):307-310.
45. Knuchel M, Ackermann M., Muller HK., Kihm U. An ELISA for detection of antibodies against porcine epidemic diarrhoea virus (PEDV) based on the specific solubility of the viral surface glycoprotein[J]. Veterinary Microbiology,1992,32(2): 117-134.
46. Kim L, Hayes J, Lewis P, Parwani AV, Chang KO, Saif LJ. Molecular characterization and pathogenesis of transmissible gastroenteritis virus(TGEV) and porcine respiratory coronavirus(PRCV) field isolates cocirculating in a swine herd[J]. Arch Virol,2000,145(6):1133-1147.
47. Kim L, Hayes J, Lewis P, Parwani AV, Chang KO, Saif LJ. Molecular characterization and pathogenesis of transmissible gastroenteritis coronavirus (TGEV) and porcine respiratory coronavirus (PRCV) field isolates co-circulating in a swine herd[J]. Arch Virol,2000,145(6):1133-1147.
48. Kim B, Chae C. In situ hybridization for the detection of transmissible gastroenteritis virus in pigs and comparison with other methods[J]. Can J Vet Res,2001,65(1): 33-37.
49. Kim SY, Song DS, Park BK. Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT-PCR[J]. J Vet Diagn Invest, 2001,13:516-520.
50. Kocherhans R, Bridgen A, Ackermann M, Tobler K. Completion of the porcine epidemic diarrhoea coronavirus (PEDV) genome sequence[J]. Virus Genes,2001, 23(2):137-144.
51. Kim SY, Song DS, Park BK..Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT-PCR[J]. J Vet Diagn Invest, 2001,13:516-520.
52. Kim O, Chae C. Experimental Infection of Piglets with a Korean Strain of Porcine Epidemic Diarrhoea Virus[J]. Journal of Comparative Pathology,2003,129(1): 55-60.
53. Kwonil Jung, Junghyum Kim, Okjin Kim, Bongtae Kim, Chanhee Chae. Differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in formalin-fixed paraffin-embedded tissues by multiplex RT-nested PCR and comparison with in situ hybridization[J]. Journal of Virological Methods,2003,108(1):41-47
54. Kwonil Jung, Chanhee Chae. RT-PCR-based dot blot hybridization for the detection and differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in fecal samples using a non-radioactive digoxigenin cDNA probe[J]. Journal of Virological Methods,2005,123(2):141-146.
55. Kim SH, Kim IJ, Pyo HM, Tark DS, Song JY, Hyun BH. Multiplex real-time RT-PCR for the simultaneous detection and quantification of transmissible gastroenteritis virus and porcine epidemic diarrhea virus[J]. J Virol Methods,2007,146(1-2): 172-177.
56. Kang JH, Kwon DH, Chung TW, Kim YD, Lee HG, Kim JW, Choe IS, Kim KW, Lim JS, Song EY, Kim CH. Development of a simple and rapid immunochromatographic strip test for diarrhea-causative porcine rotavirus in swine stool[J]. J Virol Methods, 2007,146(1-2):74-79.
57. Kim HJ, Park SI, Ha TP, Jeong YJ, Kim HH, Kwon HJ, Kang MI, Cho KO, Park SJ. Detection and genotyping of Korean porcine rotaviruses[J]. Vet Microbiol,2010.
58. Lanza I, Rubio P, Munoz M, Carmenes P. Comparison of a monoclonal antibody capture ELISA (MACELISA) to indirect ELISA and virus neutralization test for the serodiagnosis of transmissible gastroenteritis virus[J]. J Vet Diagn Invest,1993,5(1): 21-25.
59. Liu C, Kokuho T, Kubota T, Watanabe S, Inumaru S,Yokomizo Y, Onodera T:A serodiagnostic ELISA using recombinant antigen of swine transmissible gastroenteritis virus nucleoprotein[J]. J Vet Med Sci,2001,63(11):1253-1256.
60. Li D, Gu AZ, Yang W, He M, Hu XH, Shi HC. An integrated cell culture and reverse transcription quantitative PCR assay for detection of infectious rotaviruses in environmental waters[J]. J Microbiol Methods,2010.
61. Lee DK, Park CK, Kim SH, Lee C. Heterogeneity in spike protein genes of porcine epidemic diarrhea viruses isolated in Korea[J]. Virus Res,2010,149(2):175-182.
62. Martella V, Ciarlet M, Banyai K, Lorusso E, Cavalli A, Corrente M, Elia G, Arista S, Camero M, Desario C, Decaro N, Lavazza A, Buonavoglia C. Identification of a novel VP4 genotype carried by a serotype G5 porcine rotavirus strain[J]. Virology, 2006,346(2):301-311.
63. Min BS, Noh YJ, Shin JH, Baek SY, Min KI, Ryu SR, Kim BG, Park MK, Choi SE, Yang EH, Park SN, Hur SJ, Ahn BY. Assessment of the quantitative real-time polymerase chain reaction using a cDNA standard for human group A rotavirus[J]. J Virol Methods,2006,137(2):280-286.
64. Martelli P, Lavazza A, Nigrelli AD, Merialdi G, Alborali LG, Pensaert MB. Epidemic of diarrhoea caused by porcine epidemic diarrhoea virus in Italy[J]. Vet Rec,2008, 162(10):307-310.
65. Martella V, Colombrita D, Lorusso E, Draghin E, Fiorentini S, De Grazia S, Banyai K, Ciarlet M, Caruso A, Buonavoglia C. Detection of a porcine-like rotavirus in a child with enteritis in Italy[J]. J Clin Microbiol,2008,46(10):3501-3507.
66. Matthijnssens J, Ciarlet M, Heiman E, Arijs I, Delbeke T, McDonald SM, Palombo EA, Iturriza-Gomara M, Maes P, Patton JT, Rahman M,Van Ranst M. Full genome-based classification of rotaviruses reveals a common origin between human Wa-Like and porcine rotavirus strains and human DS-1-like and bovine rotavirus strains[J]. J Virol,2008,82(7):3204-3219.
67. Mulherin E, Bryan J, Beltman M, OGrady L, Pidgeon E, Garon L, Lloyd A, Bainbridge J, O'Shea H, Whyte P, Fanning S. Molecular characterisation of a bovine-like rotavirus detected from a giraffe [J]. BMC Vet Res,2008,4:46.
68. Mukherjee A, Dutta D, Ghosh S, Bagchi P, Chattopadhyay S, Nagashima S, Kobayashi N, Dutta P, Krishnan T, Naik TN, Chawla-Sarkar M. Full genomic analysis of a human group A rotavirus G9P[6] strain from Eastern India provides evidence for porcine-to-human interspecies transmission[J]. Arch Virol,2009,154(5): 733-746.
69. Miyazaki A, Fukuda M, Kuga K, Takagi M, Tsunemitsu H. Prevalence of Antibodies against Transmissible Gastroenteritis Virus and Porcine Respiratory Coronavirus among Pigs in Six Regions in Japan[J]. J Vet Med Sci,2010.
70. Nakagomi O, Nakagomi T, Akatani K, Ikegami N. Identification of rotavirus genogroups by RNA-RNA hybridization[J]. Mol Cell Probes,1989,3(3):251-261.
71. Nakagomi O, Nakagomi T. Molecular evidence for naturally occurring single VP7 gene substitution reassortant between human rotaviruses belonging to two different genogroups[J]. Arch Virol,1991,119(1-2):67-81.
72. Nakagomi O,Nakagomi T. Genomic relationships among rotaviruses recovered from various animal species as revealed by RNA-RNA hybridization assays[J]. Res Vet Sci, 2002,73(3):207-214.
73. Oh JS, Song DS, Yang JS, Song JY, Moon HJ, Kim TY, Park BK. Comparison of an enzyme-linked immunosorbent assay with serum neutralization test for serodiagnosis of porcine epidemic diarrhea virus infection[J]. J Vet Sci,2005,6(4):349-52.
74. Pritchard GC, Panton DJ, Wibberley G, Ibata G. Transmissible gastroenteritis and porcine epidemic diarrhea in Britain[J]. Vet Rec,1999,144:616-618.
75. Paton D, Ibata G, Sands J, McGoldrick A. Detection of transmissible gastroenteritis virus by RT-PCR and differentiation from porcine respiratory coronavirus[J]. J Virol Methods,1997,66(2):303-309.
76. Pritchard GC, Paton DJ, Wibberley G, Ibata G.transmissible gastroenteritis and porcine epidemic diarrhoea in Britain[J]. J Vet Res,1999,144:616-618.
77. Prabha S, Verghese S. Detection of porcine rotavirus from tissue and faecal specimens[J]. Indian J Med Microbiol,2009,27(2):149-152.
78. Park JE, Cruz DJ, Shin HJ. Trypsin-induced hemagglutination activity of porcine epidemic diarrhea virus[J]. Arch Virol,2010,155(4):595-599.
79. Riffault S, Grosclaude J, Vayssier M, Laude H, Charley B. Reconstituted coronavirus TGEV virosomes lose the virus ability to induce porcine interferon-alpha production[J]. Vet Res,1997,28(1):77-86
80. Rodak L, Smid B, Nevorankova Z, Valicek L, Smitalova R. Use of monoclonal antibodies in blocking ELISA detection of transmissible gastroenteritis virus in faeces of piglets [J].J Vet Med B Infect Dis Vet Public Health,2005,52(3):105-11.
81. Rodak L, Smitalova R, Smid B, Nevorankova Z, Valicek L, Psikal I. Comparing electron microscopy and a competitive blocking ELISA in the detection of rotaviruses in porcine faeces[J]. Vet J,2009.
82. Saif LJ, Terrett LA, Miller KL, Cross RF. Serial propagation of porcine group C rotavirus (pararotavirus) in a continuous cell line and characterization of the passaged virus[J]. J Clin Microbiol,1988,26:1277-1282.
83. Sirinarumitr T, Siddle S, Grahare F. Porcine transmissible gastroenteritis virus induce apoptosis inswine testes cell culture[J]. Archives of Virology,1998,143(12): 2471-2485.
84. Sestak K, Zhou Z, Shoup DI, Saif LJ. Evaluation of the baculovirus-expressed S glycoprotein of transmissible gastroenteritis virus(TGEV) as antigen in a competition ELISA to differentiate porcine respiratory coronavirus from TGEV antiboies in pig[J]. J Vet Diagn Invest,1999,11(3):205-14.
85. Santos N, Lima RC, Nozawa CM, Linhares RE, Gouvea V. Detection of porcine rotavirus type G9 and of a mixture of types G1 and G5 associated with Wa-like VP4 specificity:evidence for natural human-porcine genetic reassortment[J]. J Clin Microbiol,1999,37(8):2734-2736.
86. Sestak K, Zhou Z, Shoup DI, Saif LJ. Evaluation of the baculovirus-expressed S glycoprotein of transmissible gastroenteritis virus (TGEV) as antigen in a competition ELISA to differentiate porcine respiratory coronavirus from TGEV antibodies in pigs[J]. J Vet Diagn Invest,1999,11(3):205-214.
87. Song DS, Kang BK, Oh JS, Ha GW, Yang JS, Moon HJ, Jang YS, Park BK. Multiplex reverse transcription-PCR for rapid differential detection of porcine epidemic diarrhea virus,transmissible gastroenteritis,and porcine group A rotavirus[J]. J Vet Diagn Invest,2006,18:278-281.
88. Sasirekha Ramani, Miren Iturriza-Gomara, Atanu Kumar Jana, Kurien Anil Kuruvilla, James J. Gray, David W. Brown, Gagandeep Kang. Whole genome characterization of reassortant G10P[11] strain (N155) from a neonate with symptomatic rotavirus infection:Identification of genes of human and animal rotavirus origin[J]. Journal of Clinical Virology,2009,45(3):237-244.
89. Sergeev OV. Porcine epidemic diarrhea[J]. Vopr Virusol,2009,54(2):4-8.
90. Theil KW, Saif LJ, Bohl EH, Agnes AG, Kohler EM. Concurrent porcine rotaviral and transmissible gastroenteritis viral infections in a three-day-old conventional pig[J]. Am J Vet Res,1979,40(5):719-721.
91. Vemulapalli R, Gulani J, Santrich C. A real-time TaqMan RT-PCR assay with an internal amplification control for rapid detection of transmissible gastroenteritis virus in swine fecal samples[J]. J Virol Methods,2009,162(1-2):231-235.
92. Woode GN, Bridger J, Hall GA, Jones JM, Jackson G The isolation of reovirus-like agents (rota-viruses) from acute gastroenteritis of piglets[J]. J Med Microbiol,1976, 9:203-209.
93. Wyatt RG, James HD Jr, Pittman AL, Hoshino Y, Greenberg HB, Kalica AR, Flores J, Kapikian AZ. Direct isolation in cell culture of human rotaviruses and their characterization into four serotypes[J]. J Clin Microbiol,1983,18(2):310-7.
94. Ward RL, Nakagomi O, Knowlton DR, McNeal MM, Nakagomi T, Clemens JD, Sack DA, Schiff GM. Evidence for natural reassortants of human rotaviruses belonging to different genogroups[J]. J Virol,1990,64(7):3219-3225
95. Wang C, Chen J, Shi H, Qiu HJ, Xue F, Liu S, Liu C, Zhu Y, Almazan F, Enjuanes L, Feng L. Rapid differentiation of vaccine strain and Chinese field strains of transmissible gastroenteritis virus by restriction fragment length polymorphism of the N gene[J]. Virus Gene,2010,10(4):81-88
96. Wang C, Chen J, Shi H, Qiu H, Xue F, Liu C, Zhu Y, Liu S, Almazan F, Enjuanes L, Feng L. Molecular characterization of a Chinese vaccine strain of transmissible gastroenteritis virus:mutations that may contribute to attenuation[J]. Virus Gene, 2010,40(3):4030-409.
97. Wang YH, Kobayashi N, Nagashima S, Zhou X, Ghosh S, Peng JS, Hu Q, Zhou DJ, Yang ZQ. Full genomic analysis of a porcine-bovine reassortant G4P[6] rotavirus strain R479 isolated from an infant in China[J]. J Med Virol,2010,82(6):1094-1102.
98. Xi-Lin Hou, Li-Yun Yu, Jianzhu Liu. Development and evaluation of enzyme-linked immunosorbent assay based on recombinant nucleocapsid protein for detection of porcine epidemic diarrhea (PEDV) antibodies[J]. Veterinary Microbiology,2007, 123(1-3):86-92