新型聚维酮碘对牛传染性鼻气管炎病毒灭活效果试验
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摘要
甲醛是广泛应用的病毒灭活剂,但其缺点也很明显,如刺激性强、破坏免疫原性、灭活不彻底等,因此需要寻找一种可以替代传统灭活剂甲醛的新型灭活剂。通过细胞盲传三代、无菌检验和台盼蓝染色等方法,结合不同灭活浓度与灭活时间双因素来筛选聚维酮碘与甲醛灭活牛传染性鼻气管炎病毒的最优条件,同时,向MDBK细胞中接种不同浓度的聚维酮碘和甲醛观察是否对MDBK细胞具有毒性和刺激性。结果表明10%的聚维酮碘24 h时可彻底灭活传染性鼻气管炎病毒,0.3%的甲醛48 h时可彻底灭活牛传染性鼻气管炎病毒,0.5%的甲醛对细胞具有强烈刺激导致绝大多数细胞死亡,而浓度为15%的聚维酮碘对MDBK细胞仍无刺激,直到浓度高达20%时才对细胞有了较大刺激,说明聚维酮碘无刺激性和毒性、安全性更高。为病毒灭活疫苗的理想灭活剂提供了新的方向,并为新型灭活剂的研发奠定基础。
Formaldehyde was a widely used virus inactivator,but its disadvantages were also obvious,such as strong irritant,destructive immunogenicity,incomplete inactivation,so it was urgent to look for a new type of formaldehyde which could replace the traditional formaldehyde inactivated.Through the blind passage of cells for three generations,sterility testing,and trypan blue staining,combining different inactivation concentration and inactivation time to screen the optimal conditions for povidone-iodine and formaldehyde inactivation of infectious rhinotracheitis virus.At the same time,inoculation of MDBK cells with various concentrations of povidone iodine and formaldehyde was observed to be toxic and irritant to MDBK cells.The results showed that 10% povidoneiodine completely inactivated infectious rhinotracheitis virus at 24 h,0.3% formaldehyde completely inactivated infectious rhinotracheitis virus at 48 h,and 0.5% formaldehyde strongly irritated cells resulting in the most dead cells,but 15% povidone-iodine had still no stimulation to MDBK cells,until the concentration of up to 20% of the cells had a greater stimulus,indicating povidone-iodine non-irritating,toxic and had more security.It provided a new direction for the ideal inactivators of virus inactivated vaccines,and laid the foundation for the development of new inactivators.
Formaldehyde was a widely used virus inactivator,but its disadvantages were also obvious,such as strong irritant,destructive immunogenicity,incomplete inactivation,so it was urgent to look for a new type of formaldehyde which could replace the traditional formaldehyde inactivated.Through the blind passage of cells for three generations,sterility testing,and trypan blue staining,combining different inactivation concentration and inactivation time to screen the optimal conditions for povidone-iodine and formaldehyde inactivation of infectious rhinotracheitis virus.At the same time,inoculation of MDBK cells with various concentrations of povidone iodine and formaldehyde was observed to be toxic and irritant to MDBK cells.The results showed that 10% povidoneiodine completely inactivated infectious rhinotracheitis virus at 24 h,0.3% formaldehyde completely inactivated infectious rhinotracheitis virus at 48 h,and 0.5% formaldehyde strongly irritated cells resulting in the most dead cells,but 15% povidone-iodine had still no stimulation to MDBK cells,until the concentration of up to 20% of the cells had a greater stimulus,indicating povidone-iodine non-irritating,toxic and had more security.It provided a new direction for the ideal inactivators of virus inactivated vaccines,and laid the foundation for the development of new inactivators.
引文
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[9]黄建飞,吴彻,洪海彬,等.聚维酮碘口服液对黄羽肉鸡肠道菌群及肠道组织结构影响的研究[J].中国家禽,2016,38(17):21-25.
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[12]雷元元,何生虎,郭亚男,等.甲醛灭活牛支原体的条件探究[J].黑龙江畜牧兽医,2015(21):36-39+287.
[13]刘全离.聚维酮碘体外抑菌效果试验观察[J].中国消毒学杂志,2008(1):28-29.
[14]胡兆勇,屈安,丛新德.聚维酮碘的临床应用[J].中国医院药学杂志,1998,18(7):307-308.
[15]钱毅春,攀树理,尹幸念,等.碘伏消毒液的毒性研究[J].中国消毒学杂志,2004,21(2):128-129.
[16]杨美玲,张紫虹,胡永成,等.一种聚维酮碘消毒液的毒理学安全性评价[J].中国卫生检验杂志,2007,17(7):1286-1288.
[17]周雯,唐慧,李慧,等.某种碘伏消毒剂的毒性试验研究[J].预防医学文献信息志,2005,10(3):313-315.
[18]陈明中,吴心勤,蔺红光,等.某种消毒液的毒性试验[J].预防医学论坛,2006,12(3):307-309.
[19]赵生友,姜晓春,王艳,等.四种消毒剂的急性毒性试验和微核试验[J].研究简报,2004,16(3):178-180.
[20]李继昌,丛霞,符春艳,等.聚维酮碘消毒剂的急性毒性试验[J].黑龙江畜牧兽医,1998(5):48.
[21]夏勇,魏兰芬,高筱萍,等.聚维酮碘消毒剂毒性的试验研究[J].中国消毒学杂志,2005,22(3):263-267.
[22]黄晓波,夏涛,彭明军,等.复方聚维酮碘消毒液的毒性研究[J].公共卫生与预防医学,2007,18(1):21-23.
[23]周立国,程灵,邵雪熹,等.碘伏消毒剂活力碘的毒性测定[J].中国消毒学杂志,1993,10(3):157-161.
[24]蒋莉,袁庆霞,武雪冰,等.聚维酮碘对脊髓灰质炎病毒杀灭效果及影响因素的观察[J].中国消毒学杂志,2005(4):368-370.
[25]张伟,张丽娜,陈亚双,等.聚维酮碘的临床应用[J].黑龙江中医药,2016,45(4):62-63.
[2]孙建滨.大鲵虹彩病毒灭活方法比较与疫苗制备技术研究[D].武汉:华中农业大学,2013.
[3]徐守振,尹燕博,王新.动物疫苗中常用抗原灭活剂的研究进展[J].中国畜牧兽医,2010,37(9):162-167.
[4]姜平.兽医生物制品学[M].2版.北京:中国农业出版社,2006.
[5]IgaN,Otsuka A,Iwata M,et al.Generalize dmorphe a with preceding severe pain and coexis tent early primary biliary cirrhosis[J].EJ D,2015,25(4):365-366.
[6]Deng C,Hu C,Wang L,et al.Serological comparative proteomic sanalys is of mitoc hond ria lautoanti body negative and positive primary biliary cirrhosis[J].Ele c Trop Horesis,2015,36(14):1588-1595.
[7]Sharon D,Ma s on A L.Role of novel retroviruses in chronic live rd is ease:assessing the link of human bet are trovirus with primary biliary cirrhosis[J].Current Infectious Disease Reports,2015,17(2):460.
[8]安多福研制出创新型非抗生素类新兽药--“万金水”聚维酮碘口服液[J].广东经济,2015(10):96.
[9]黄建飞,吴彻,洪海彬,等.聚维酮碘口服液对黄羽肉鸡肠道菌群及肠道组织结构影响的研究[J].中国家禽,2016,38(17):21-25.
[10]周慧芳,洪敏,王梦甜,等.聚维酮碘溶液对子宫内膜细胞的灭活作用研究[J].浙江医学,2016,38(24):1973-1976.
[11]梁宏儒,高佳滨,李安,等.牛传染性鼻气管炎的分离鉴定和灭活条件的优化[J].黑龙江八一农垦大学学报,2013,25(6):35-38.
[12]雷元元,何生虎,郭亚男,等.甲醛灭活牛支原体的条件探究[J].黑龙江畜牧兽医,2015(21):36-39+287.
[13]刘全离.聚维酮碘体外抑菌效果试验观察[J].中国消毒学杂志,2008(1):28-29.
[14]胡兆勇,屈安,丛新德.聚维酮碘的临床应用[J].中国医院药学杂志,1998,18(7):307-308.
[15]钱毅春,攀树理,尹幸念,等.碘伏消毒液的毒性研究[J].中国消毒学杂志,2004,21(2):128-129.
[16]杨美玲,张紫虹,胡永成,等.一种聚维酮碘消毒液的毒理学安全性评价[J].中国卫生检验杂志,2007,17(7):1286-1288.
[17]周雯,唐慧,李慧,等.某种碘伏消毒剂的毒性试验研究[J].预防医学文献信息志,2005,10(3):313-315.
[18]陈明中,吴心勤,蔺红光,等.某种消毒液的毒性试验[J].预防医学论坛,2006,12(3):307-309.
[19]赵生友,姜晓春,王艳,等.四种消毒剂的急性毒性试验和微核试验[J].研究简报,2004,16(3):178-180.
[20]李继昌,丛霞,符春艳,等.聚维酮碘消毒剂的急性毒性试验[J].黑龙江畜牧兽医,1998(5):48.
[21]夏勇,魏兰芬,高筱萍,等.聚维酮碘消毒剂毒性的试验研究[J].中国消毒学杂志,2005,22(3):263-267.
[22]黄晓波,夏涛,彭明军,等.复方聚维酮碘消毒液的毒性研究[J].公共卫生与预防医学,2007,18(1):21-23.
[23]周立国,程灵,邵雪熹,等.碘伏消毒剂活力碘的毒性测定[J].中国消毒学杂志,1993,10(3):157-161.
[24]蒋莉,袁庆霞,武雪冰,等.聚维酮碘对脊髓灰质炎病毒杀灭效果及影响因素的观察[J].中国消毒学杂志,2005(4):368-370.
[25]张伟,张丽娜,陈亚双,等.聚维酮碘的临床应用[J].黑龙江中医药,2016,45(4):62-63.