新生牛血清病毒检测
摘要
现代生物技术一般认为包括基因工程技术、细胞工程技术、酶工程技术和发酵工程技术,而这些技术的发展几乎都与细胞培养有密切关系,特别是在医药领域的发展,细胞培养更具有特殊的作用和价值。细胞培养在整个生物技术产业的发展中起到了很关键的核心作用。细胞培养的发展,培养基的质量又是关键,而培养基的主要成份中动物血清对细胞的生长繁殖发挥着重要甚至是难以替代的作用。在动物血清的应用中牛血清又是最为广泛的,所以牛血清是医药生物技术产品中重要的原材料之一。保证牛血清质量也是促进生物制品质量提高的重要环节。
新生牛血清作为生物制品,尤其是疫苗的生产主要原料,其质量直接关系到疫苗质量,因此对新生牛血清中病毒检测尤为重要,如：牛腹泻病毒、牛腺病毒、牛细小病毒、呼肠孤病毒、狂犬病病毒、牛副流感病毒等,检查方法一般为细胞培养法、血吸附法检测法及免疫荧光抗体检查法。
本文主要研究细胞培养法、血吸附法检测法和免疫荧光抗体检查法检测新生牛血清的检测时间及检测准确度,为疫苗生产提供质量保证,使新生牛血清病毒检测方法完全满足GMP要求。
新生牛血清系从14小时内未进食的新生牛采血分离血清,经除菌过滤后制成,牛血清生产过程中不得任意添加其他物质成分。新生牛血清用于生产前应逐批进行蛋白质含量、血红蛋白、大肠杆菌噬菌体、渗透压摩尔浓度、无菌检查、支原体检查、细菌内毒素检查以及病毒检查,本文仅讨论病毒检查的内容,对30组供试品使用细胞培养法及免疫荧光抗体检查法进行检测,检测结果应为阴性,符合要求。
Generally, modern biological technology has been included genetic engineering, cellular engineeering, enzyme engineering technology and fermentation engineering technology, the developpment of these techniques almost all have relationship with the technology of cell cultu-re, es- pecially, the cell culture has more special action and value in development of medical field. The technology of cell culture plays an essential role in the development of biotechnology industry. Development of the cell culture technology, the quality of the medium is pivotal, but the main ingredients of animal medium for cells to grow breeding is the serum which play an important even irreplaceable role. In the application of animal serum the bovine serum is the most extens- ive, so the bovine serum is one of the important raw materials for the pharmaceuti-cal biotechnology product. Ensuring the quality of bovine serum is also an important part of promoting the quality of biological products.
The newborn bovine serum as the raw materials of the biological products, especially the production of vaccine, its quality related to production and quality of the vaccine. So the virus detection of the newborn bovine serum is particularly important. Such as:Bovine viral diarrhea virus(BVDV), Bovine adenovirus(BAV), Bovine Parvo virus(BPV), Reovirus, Rabies virus(RV) and Bovine parainfluenza virus(BPIV). etc must be detected, the method of the detection is the cell culture, blood adsorption assay and immune fluorescent antibody(IFA).
This paper mainly studies the cell culture, blood adsorption assay and immune fluorescent antibody methods to test of testing time and detecting accuracy of newborn bovine serum. Make the detection method of the virus from newborn bovine serum meeting completely the require-ments of GMP.
The newborn bovine serum is that take from the newborn cattle without food below 14 hours, and made after aseptic filtration, the process of producting the bovine serum can not add other material composition arbitrarily. Before the newborn bovine serum which used in the pro-duction should detect batch-by-batch for protein content, hematoglobin, escherichia coli phage, osmotic pressure molarity, asepsis check, mycoplasma inspection, bacterial endotoxin check and a virus-check. This paper discussed the contents of virus checking only in the newborn bovine serum that used the 30 batch of samples by the methods of cell culture and immune fluorescent antibody. The test results should be negative, and meets the requirement.
新生牛血清作为生物制品,尤其是疫苗的生产主要原料,其质量直接关系到疫苗质量,因此对新生牛血清中病毒检测尤为重要,如：牛腹泻病毒、牛腺病毒、牛细小病毒、呼肠孤病毒、狂犬病病毒、牛副流感病毒等,检查方法一般为细胞培养法、血吸附法检测法及免疫荧光抗体检查法。
本文主要研究细胞培养法、血吸附法检测法和免疫荧光抗体检查法检测新生牛血清的检测时间及检测准确度,为疫苗生产提供质量保证,使新生牛血清病毒检测方法完全满足GMP要求。
新生牛血清系从14小时内未进食的新生牛采血分离血清,经除菌过滤后制成,牛血清生产过程中不得任意添加其他物质成分。新生牛血清用于生产前应逐批进行蛋白质含量、血红蛋白、大肠杆菌噬菌体、渗透压摩尔浓度、无菌检查、支原体检查、细菌内毒素检查以及病毒检查,本文仅讨论病毒检查的内容,对30组供试品使用细胞培养法及免疫荧光抗体检查法进行检测,检测结果应为阴性,符合要求。
Generally, modern biological technology has been included genetic engineering, cellular engineeering, enzyme engineering technology and fermentation engineering technology, the developpment of these techniques almost all have relationship with the technology of cell cultu-re, es- pecially, the cell culture has more special action and value in development of medical field. The technology of cell culture plays an essential role in the development of biotechnology industry. Development of the cell culture technology, the quality of the medium is pivotal, but the main ingredients of animal medium for cells to grow breeding is the serum which play an important even irreplaceable role. In the application of animal serum the bovine serum is the most extens- ive, so the bovine serum is one of the important raw materials for the pharmaceuti-cal biotechnology product. Ensuring the quality of bovine serum is also an important part of promoting the quality of biological products.
The newborn bovine serum as the raw materials of the biological products, especially the production of vaccine, its quality related to production and quality of the vaccine. So the virus detection of the newborn bovine serum is particularly important. Such as:Bovine viral diarrhea virus(BVDV), Bovine adenovirus(BAV), Bovine Parvo virus(BPV), Reovirus, Rabies virus(RV) and Bovine parainfluenza virus(BPIV). etc must be detected, the method of the detection is the cell culture, blood adsorption assay and immune fluorescent antibody(IFA).
This paper mainly studies the cell culture, blood adsorption assay and immune fluorescent antibody methods to test of testing time and detecting accuracy of newborn bovine serum. Make the detection method of the virus from newborn bovine serum meeting completely the require-ments of GMP.
The newborn bovine serum is that take from the newborn cattle without food below 14 hours, and made after aseptic filtration, the process of producting the bovine serum can not add other material composition arbitrarily. Before the newborn bovine serum which used in the pro-duction should detect batch-by-batch for protein content, hematoglobin, escherichia coli phage, osmotic pressure molarity, asepsis check, mycoplasma inspection, bacterial endotoxin check and a virus-check. This paper discussed the contents of virus checking only in the newborn bovine serum that used the 30 batch of samples by the methods of cell culture and immune fluorescent antibody. The test results should be negative, and meets the requirement.
引文
[1]中国生物制品生产用主要原辅材料质控标准,2000年版.北京：中国生物制品标准化委员会,2000
[2]中华人民共和国药典2010年版,第三部,附录113,国家药典委员会编.北京：中国医药科技出版社,2010
[3]尹红章,李秀华,曾麟,等.应用细胞倍增时间测定新生牛血清的质量[J].中国生物制品学杂志,1993,6(3),116-117
[4]乔自林,冯玉萍,李明生,等.我国新生牛血清的生产现状[J].西北民族大学学报,30(76)：43-45
[5]宋立荣,李思经.谈新生牛血清的质量控制[J].黑龙江畜牧兽医,2008(10)92-94
[6]张浩.谈牛血清生产工艺设计的指导原则[J].山西科技,2002(2)：51-52
[7]张浩,刘文静,张晓杰.粗制牛血清的质量控制[J].科技情报开发与经济,2001,11(6)：128-129
[8]刘文静.保证牛血清质量促进生物制品品质的提高[J].科技情报开发与经济,2001,11(5)：58-59
[9]ISO 14644-1:1999(E), Part 1,3. Geneva, Switzerland:International Organization for Standardization. 1999
[10]中华人民共和国药典2010年版,第二部.国家药典委员会编.北京：中国医药科技出版社,2010
[11]熊永忠,苏雅君,万文英.如何做好新生犊牛血清的生产管理和质量监控[J].畜牧兽医科技信息,2005.06
[12]用动物细胞体外培养生产生物制品规程,Geneva, Switzerland:World Health Organization,2003
[13]中国生物制品标准化委员会.中国生物制品主要原辅材料质控标准[M].北京：化学工业出版社,2000.57
[14]董关木.细胞培养用牛血清现状与进展[J].中国生物制品学杂志,2007,20(9)：697-700
[15]陈波等.牛血清中牛腹泻病毒牛肾细胞直接病变检测方法的建立[J].中国生物制品学杂志,2009,22(8)：819-822
[16]Greiser-Wilk I, Grummer B, Moennig V. Bovine viral diarrhea eradication and control programmes in Europe. Biologicals,2003,31 (2):113-118
[17]Lopez OJ, Osorio FA, Donis RO. Rapid detection of bovine viral diarrhea virus by polymerase chain reaction. J Clin Microbiol,1991,29(3):578-582
[18]Sasaki T, Harasawa R, Shintani M, et al. Application of PCR for detection of mycoplasma DNA and pestivirus RNA in human live viral vaccines. Biologicals,1996,24(4):371-375
[19]Bolin SR, Ridpath JF, Black J, et al. Survey of cell lines in the American Type Culture Collection for bovine viral diarrhea virus. JVirol Methods,1994,48(2-30:211-221
[20]Harasawa R, Tomiyama T. Evidence of pestivirus RNA in humanvirus vaccines. J Clin Microbiol,1994, 32(6):1604-1605
[21]Wilks CR, Abrahain G, Blackmore DK. Bovine pestivirus and human infection. The Lancet,1989,1 (8629):107
[22]Makoschey B, Gelder P, Keijsers V, et al. Bovine viral diarrhea virus antigen in fetal calf serum batches and consequences of such contamination for vaccine production. Biologicals,2003,31 (3):203-208
[23]黄瑜,甘孟侯,刘尚高,等.中国畜牧兽医学会家畜传染病学分会第五次学术研讨会论文集,1993,247～249
[24]李文刚,甘孟侯.中国畜牧兽医学会家畜传染病学分会第6次学术研讨会论文集,1995,168
[25]Klein M.el al, Proc, Soc. Expll. Biol.&Med.1959,102,1-4
[26]Belak D.E. Am.J.Vet.Res.1978,39,12,1904-1906
[27]Wigand, R.et al, Intervirnary,1982,18,169-176
[28]Coria, M.F.and A.W.MeClurkin, Am.J.Vet Res.1978,39.12,1975-1976
[29]Mohany, S.B.et al, Velerinary Viro-logy Balliere Tindall. London.1981
[30]Shiu-lok Hu et al, Journal of Virology,1984,51 (3),880-883
[31]Belak, S.et al, Veterinary Microbiology,1986,12,87-91
[32]Baber, D.I, Research in Veterinary,1981,31,69-75
[33]Coria,M.F, Am.J.Vet.Res.1978,39,12,1904-1906
[34]黎炜.牛细小病毒感染[J].黑龙江畜牡兽医,1989,10：35-36
[35]蔡翼虎,王玉芳,严雪仙.狂犬病病毒检测方法的研究进展[J].浙江畜牧兽医,2009,2：12-13
[36]Alexander Bukreyev, Mario HS, Brian RM, et al. Nonsegmented negative-strand viruses as vaccine vectors. Journal of Virology,2006,80(21):10293-10306
[37]Reisinger, Heddleston, Manthei, Isolation of bovine parainfluenza-3 virus in chick embryos. J am vet med assoc,1959,135:147
[38]刘鹏,侯喜林,周玉龙,等.牛副流感病毒3型的分离鉴定[J].微生物学通报,2009,36(9)：1384-1389
[39]David PG, Robert EW, Lee MS, et al. A bovine parainfluenza virus type3 vaccine is safe and immunogenic in early infancy. The Journal of Infectious Diseases,2005,191:1116-1122
[40]Horwood PF, Gravel JL, Mahony TJ, et al. Identification of two distinct bovine parainfluenza virus type 3 genotypes.The Journal of Gen Virol,2008,89:1643-1648
[41]常继涛,李鑫,张亚科,等.犊牛腹泻粪样中牛呼肠孤病毒的分离鉴定[J].中国预防兽医学报,2008-9,30(9),711-715
[42]Knipe, David M, Howley. Fields Virology 4th Edition [M]. Lippincott Willians and Wilkins,2001, 2636-2663
[43]L William Cashdollar, Richard A Chmelo, Jon R Wiener, et al.Sequences of the S1 genes of the three serotypes of reovirus [J]. Proc Natl Acad Sci,1985,82:24-28
[44]曾智勇,郭万柱,徐志文等.仔猪腹泻粪样中猪呼肠孤病毒的分离鉴定[J].畜牧兽医学报,2007, 38：574-580
[45]殷震,刘景华.动物病毒学[M].第2版,北京：科学出版社,1997,538-578
[46]端青,朱虹,杨怡等.呼肠病毒在SARS患者中分离与初步鉴定[J].科学通报,2003,48(13)：1369-1372
[47]Tillot son J R, Lerner A M. Reovirus type 3 associated with fatal pneumonia [J]. The New England Journal of Medicine,1967,276 (19):1060-1063
[48]朱虹,何君,何诚,等.新分离呼肠病毒的致病性及其与SARS相关性的实验动物研究[J].军事医学科学院院刊,2004,28(3)：295-297
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[52]梁名践,蒋茨蕾,鲁宏,等.新生新生牛血清在精制地鼠肾细胞狂犬疫苗组织培养中对肾细胞保护作用的观察[J].中国媒介生物学及控制杂志,2001,12(2)：157
[53]刘学平,沈劲草,魏至栋,等.牛肾原代细胞培养最适接种浓度的选择[J].甘肃科技2009,19：62
[54]陈波,陈秀珍,徐程林,等.直接病变法检测牛腹泻病毒的方法建立与验证,2005年10月1日,第八次全国生物制品学术会议论文汇编,2004-2005
[2]中华人民共和国药典2010年版,第三部,附录113,国家药典委员会编.北京：中国医药科技出版社,2010
[3]尹红章,李秀华,曾麟,等.应用细胞倍增时间测定新生牛血清的质量[J].中国生物制品学杂志,1993,6(3),116-117
[4]乔自林,冯玉萍,李明生,等.我国新生牛血清的生产现状[J].西北民族大学学报,30(76)：43-45
[5]宋立荣,李思经.谈新生牛血清的质量控制[J].黑龙江畜牧兽医,2008(10)92-94
[6]张浩.谈牛血清生产工艺设计的指导原则[J].山西科技,2002(2)：51-52
[7]张浩,刘文静,张晓杰.粗制牛血清的质量控制[J].科技情报开发与经济,2001,11(6)：128-129
[8]刘文静.保证牛血清质量促进生物制品品质的提高[J].科技情报开发与经济,2001,11(5)：58-59
[9]ISO 14644-1:1999(E), Part 1,3. Geneva, Switzerland:International Organization for Standardization. 1999
[10]中华人民共和国药典2010年版,第二部.国家药典委员会编.北京：中国医药科技出版社,2010
[11]熊永忠,苏雅君,万文英.如何做好新生犊牛血清的生产管理和质量监控[J].畜牧兽医科技信息,2005.06
[12]用动物细胞体外培养生产生物制品规程,Geneva, Switzerland:World Health Organization,2003
[13]中国生物制品标准化委员会.中国生物制品主要原辅材料质控标准[M].北京：化学工业出版社,2000.57
[14]董关木.细胞培养用牛血清现状与进展[J].中国生物制品学杂志,2007,20(9)：697-700
[15]陈波等.牛血清中牛腹泻病毒牛肾细胞直接病变检测方法的建立[J].中国生物制品学杂志,2009,22(8)：819-822
[16]Greiser-Wilk I, Grummer B, Moennig V. Bovine viral diarrhea eradication and control programmes in Europe. Biologicals,2003,31 (2):113-118
[17]Lopez OJ, Osorio FA, Donis RO. Rapid detection of bovine viral diarrhea virus by polymerase chain reaction. J Clin Microbiol,1991,29(3):578-582
[18]Sasaki T, Harasawa R, Shintani M, et al. Application of PCR for detection of mycoplasma DNA and pestivirus RNA in human live viral vaccines. Biologicals,1996,24(4):371-375
[19]Bolin SR, Ridpath JF, Black J, et al. Survey of cell lines in the American Type Culture Collection for bovine viral diarrhea virus. JVirol Methods,1994,48(2-30:211-221
[20]Harasawa R, Tomiyama T. Evidence of pestivirus RNA in humanvirus vaccines. J Clin Microbiol,1994, 32(6):1604-1605
[21]Wilks CR, Abrahain G, Blackmore DK. Bovine pestivirus and human infection. The Lancet,1989,1 (8629):107
[22]Makoschey B, Gelder P, Keijsers V, et al. Bovine viral diarrhea virus antigen in fetal calf serum batches and consequences of such contamination for vaccine production. Biologicals,2003,31 (3):203-208
[23]黄瑜,甘孟侯,刘尚高,等.中国畜牧兽医学会家畜传染病学分会第五次学术研讨会论文集,1993,247～249
[24]李文刚,甘孟侯.中国畜牧兽医学会家畜传染病学分会第6次学术研讨会论文集,1995,168
[25]Klein M.el al, Proc, Soc. Expll. Biol.&Med.1959,102,1-4
[26]Belak D.E. Am.J.Vet.Res.1978,39,12,1904-1906
[27]Wigand, R.et al, Intervirnary,1982,18,169-176
[28]Coria, M.F.and A.W.MeClurkin, Am.J.Vet Res.1978,39.12,1975-1976
[29]Mohany, S.B.et al, Velerinary Viro-logy Balliere Tindall. London.1981
[30]Shiu-lok Hu et al, Journal of Virology,1984,51 (3),880-883
[31]Belak, S.et al, Veterinary Microbiology,1986,12,87-91
[32]Baber, D.I, Research in Veterinary,1981,31,69-75
[33]Coria,M.F, Am.J.Vet.Res.1978,39,12,1904-1906
[34]黎炜.牛细小病毒感染[J].黑龙江畜牡兽医,1989,10：35-36
[35]蔡翼虎,王玉芳,严雪仙.狂犬病病毒检测方法的研究进展[J].浙江畜牧兽医,2009,2：12-13
[36]Alexander Bukreyev, Mario HS, Brian RM, et al. Nonsegmented negative-strand viruses as vaccine vectors. Journal of Virology,2006,80(21):10293-10306
[37]Reisinger, Heddleston, Manthei, Isolation of bovine parainfluenza-3 virus in chick embryos. J am vet med assoc,1959,135:147
[38]刘鹏,侯喜林,周玉龙,等.牛副流感病毒3型的分离鉴定[J].微生物学通报,2009,36(9)：1384-1389
[39]David PG, Robert EW, Lee MS, et al. A bovine parainfluenza virus type3 vaccine is safe and immunogenic in early infancy. The Journal of Infectious Diseases,2005,191:1116-1122
[40]Horwood PF, Gravel JL, Mahony TJ, et al. Identification of two distinct bovine parainfluenza virus type 3 genotypes.The Journal of Gen Virol,2008,89:1643-1648
[41]常继涛,李鑫,张亚科,等.犊牛腹泻粪样中牛呼肠孤病毒的分离鉴定[J].中国预防兽医学报,2008-9,30(9),711-715
[42]Knipe, David M, Howley. Fields Virology 4th Edition [M]. Lippincott Willians and Wilkins,2001, 2636-2663
[43]L William Cashdollar, Richard A Chmelo, Jon R Wiener, et al.Sequences of the S1 genes of the three serotypes of reovirus [J]. Proc Natl Acad Sci,1985,82:24-28
[44]曾智勇,郭万柱,徐志文等.仔猪腹泻粪样中猪呼肠孤病毒的分离鉴定[J].畜牧兽医学报,2007, 38：574-580
[45]殷震,刘景华.动物病毒学[M].第2版,北京：科学出版社,1997,538-578
[46]端青,朱虹,杨怡等.呼肠病毒在SARS患者中分离与初步鉴定[J].科学通报,2003,48(13)：1369-1372
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