布氏菌纯蛋白衍生物.doc（布氏菌纯蛋白衍生物.doc）

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1、布氏菌纯蛋白衍生物.doc（布氏菌纯蛋白衍生物.doc）布氏菌纯蛋白衍生物BushijunchundanbaiyanshengwuPurifiedProteinderivative(PPDbrucellinofBR-)本品系用布氏菌经培养、杀菌、除去菌体后的上清液制成的纯蛋白衍生物,用于布氏病的临床诊断、布氏菌苗接种对象的选择及布氏菌苗接种后机体免疫反应的监测.1基本要求生产和检定用设施、原料及辅料、水、器具、动物等应符合"凡例"的有关要求.布氏菌纯蛋白衍生物(BP-PPD)生产车间必须与其他生物制品生产车间及实验室

2、分开,原液生产全部过程,包括布氏杆菌的灭活,应在完全隔离的区域内进行,所需设备及器具均须单独设置并专用.直接用于生产的金属或玻璃等器具,应经过严格清洗及灭菌处理.2制造2.1菌种生产用菌种应符合"生物制品生产检定用菌毒种管理规程"的有关规定.2.2.1名称及来源采用猪布氏菌Ⅰ型cmcc55007菌株(S2).2.1.2种子批的建立应符合"生物制品生产检定用菌毒种管理规程"的有关规定.2.1.3种子批的传代工作种子批启开至菌体收集传代应不超过5代.2.1.4种子批菌种的检定2.1.4.1染色镜检菌体染色后镜检应为革兰阴

3、性小杆菌.2.1.4.2生化特性1:1000,three,emodinshouldbenegativeforvariationtest;itcangrowinsulfurvioletmedium;doesnotgrowinbasicfuchsinmedium;canproduceobvioushydrogensulfideinthegrowthofculture,withoutcarbondioxide.Serologicalcharacteristicsof2.1.4.3ThestrainsofBrucellare

4、ferenceserumagglutinationtitershouldreach1to800(++)above.Cleavagecharacteristicsof2.1.4.4phageItcanbecleavedbyBrucellabacteriophageWb.Preservationof2.and1.5seedbatchesFreezedriedstrainsarepreservedbelow8degrees,andliquidstrainsarepreservedbelow-70centigrade.2.2

5、dopeSeedfor2.2.1productionTheopeningstrainswereinoculatedintheliveragarmedium,37DEGCandculturedfor2to3days,accordingtothegrowthsituation,intheliveronagarslantthenpassaged1times,thegrowthofgoodlawn,availableforalargenumberoftheuseofvaccination.Culturemediumfor2.

6、2.2productionLiverextractagarmediumorotherapprovedmediumwasused.Inoculationandcultureof2.2.3strains2.2.1agoodgrowthinlargelawn,inoculationamountofliverbevelorKbottleasseed,37DEGCandculturedfor2days.Alargehepaticslopeoraliveragarbottlewasinoculatedat37daysfor2da

7、ys.2.2.4harvestandsterilizationCultureswereterminated,washedwithSodiumChloridePhysiologicalSolution,sterilizedat121Cfor30minutes,centrifugedandremovedthesupernatant,andsupernatantwascollected.Theabovesolutionshouldbeaddedtokeep3.0g/Lphenolorothersuitablepreserv

8、atives,preservedin4~8degrees,preservationperiodofnotmorethan30days.Freezingshouldbeprevented.2.2.5purificationThesupernatantwascollectedforpurification.Theproteinswereprecip