非分型流感嗜血杆菌Haps蛋白的分离纯化

目的优化非分型流感嗜血杆菌Haps蛋白质片段的分离纯化方法。方法采用盐析、透析脱盐、超滤浓缩、弱阳离子交换柱Hitrap CM Sepharose Fast Flow层析纯化HapS蛋白,优化Hap。蛋白质片段的洗脱条件,包括pH值和离子强度,测定各洗脱液样品在280nm波长处的光吸收值（D280）,用折线图显示,SDS-PAGE电泳检测分布图中处于峰值的样品,观察目的蛋白条带的出现。结果弱阳离子交换柱Hitrap CM Sepharose Fast Flow层析纯化HapS蛋白,缓冲液l洗脱液的D280分布为基线,缓冲液2洗脱液的D280分布折线图中有峰值出现,但峰有拖尾,SDS-PAGE电泳检测该洗脱峰主要为低分子量的蛋白条带;五种不同离子强度的缓冲液3洗脱液在100mmol／L NaCl离子强度时,D280的分布折线图显示有较高洗脱峰出现,SDS-PAGE电泳检测该洗脱峰有较明显的110000D的目的条带,其余离子强度下均未见明显洗脱峰出现。结论Sepharose CM FF层析柱分离纯化Haps蛋白质片段时,不同pH值的缓冲液对目的蛋白的洗脱没有明显影响,而主要影响杂蛋白的洗脱;100mmol／L NaCl离子强度的缓冲液3洗脱液获得较好的洗脱效果。     

B型流感嗜血杆菌疫苗的研究进展

B型流感嗜血杆菌(Haemophilus influenzae type b,Hib)是小儿细菌性感染的主要元凶之一。据世界卫生组织(WHO)2012年4月统计报道,2008年全世界因Hib感染死亡的5岁以下儿童人数为20.3万,而目前预防Hib最有效的措施是接种Hib疫苗。本文回顾了Hib疫苗的发展,对现有Hib结合疫苗的免疫原性、结合方式、结合策略进行综述。     

b型流感嗜血杆菌结合疫苗的安全性评价

目的:评价b型流感嗜血杆菌结合疫苗的安全性,为Hib疫苗的免疫接种推广提供依据。方法:回顾性分析自愿接种Hib疫苗的10 087名6个月龄~5周岁的婴幼儿的资料,按年龄分为6~11个月龄组(1 163名)和12~59个月龄组(8 924名);分析儿童接种后不良反应发生情况。结果:6~11个月龄组幼儿接种72 h后的局部不良反应发生率明显高于12~59个月龄组幼儿,差异具有统计学意义(P﹤0.05);6~11个月龄组幼儿接种72 h后全身不良反应发生率高于12~59个月龄组幼儿,差异无统计学意义(P﹥0.05)。结论:b型流感嗜血杆菌结合疫苗在6个月龄~5周岁的婴幼儿中使用具有较好的安全性,但在接种过程中应密切关注幼儿出现的局部反应和全身反应,并进行及时处理。     

通过CDAP活化多糖制备b型流感嗜血杆菌荚膜多糖-D蛋白疫苗初探

目的 用CDAP活化b型流感嗜血杆菌（Hib）荚膜多糖,制备b型流感嗜血杆菌荚膜多糖及D蛋白疫苗.方法 采用CDAP法在不同pH值条件下活化Hib荚膜多糖,再通过乙二酰肼（ADH）与D蛋白共价结合,制备Hib荚膜多糖与其D蛋白结合物原液,并对三批结合物原液的各项指标进行检测.获得的结合物原液免疫BALB/c小鼠,应用ELISA法检测血清中的IgG抗体水平,评价其免疫原性.结果 三批结合物原液以上检测指标均符合药典要求,衍生物中ADH含量对结合物的免疫原性有重要影响.结论 用CDAP活化工艺制备的Hib荚膜多糖-D蛋白结合物适宜制备结合疫苗,为以D蛋白为载体的Hib结合疫苗的制备及五价联苗的研制奠定实验基础.     

流感嗜血杆菌的蛋白电泳图谱分析及分型

目的 分析流感嗜血杆菌蛋白的成分及免疫原性。为流感嗜血杆菌菌苗的研制提供依据。方法 应用聚丙烯酰胺凝胶电泳分析５６株流感嗜血杆菌蛋白成分。结果 ５６株流感嗜血杆菌中不同分型流感嗜血杆菌４４株,占７９％。生物分型中,Ⅱ型为最高,占４８％,流感嗜血杆菌约有２０多条蛋白带,１１条为所有型别菌株共同具有的,其中以相对分子量为９６０００、５１０００及３１０００者含量较高,各菌株间的差异蛋白的相对分子量主要为     

312例国产b型流感嗜血杆菌结合疫苗副反应与其相关因素分析

目的 了解国产的b型流感嗜血杆菌结合疫苗副反应以及相关因素,为安全接种提供科学依据.方法 收集本辖区满1-5周岁儿童在2006年～2009年间接种b型流感嗜血杆菌结合疫苗3487例出现的312例副反应进行分析.结果 四年来,国产b型流感嗜血杆菌结合疫苗的副反应呈逐年下降趋势,与疫苗本身无关,主要原因与工作人员对家长施行的卫生宣教逐年加强和接种时气温有关,气温高时局部和全身副反应发生率高,反之,发生率低,(P＜0.05),差别有显著意义.结论 接种国产b型流感嗜血杆菌结合疫苗前的儿童要经医生严格体检,身体状态良好者方可接种,接种后对家长施行详细而到位的卫生宣教,接种时最好避开高温天气.     

B型流感嗜血杆菌感染所致疾病及其免疫预防

Ｂ型流行性感冒嗜血杆菌 (Ｈａｅｍｏｐｈｉｌｕｓｉｎｆｌｕｅｎｚａｅｔｙｐｅｂ ,Ｈｉｂ)是引起小儿严重细菌感染的主要致病菌 ,通过鼻咽部和呼吸道分泌物形成的飞沫传播 ,所致感染性疾病主要发生在 5岁以下儿童 ,病死率在发达国家为 5 %,在发展中国家可高达 4 0 %,是引起婴幼儿死亡的主要原因[1] 。由于其发病范围广 ,后果严重 ,目前已成为一大公共卫生问题。1　Ｈｉｂ感染所致疾病在未实施Ｈｉｂ疫苗免疫接种前 ,发达国家 5岁以下儿童Ｈｉｂ感染所致疾病年发病率为 2 0 / 10万～ 10 0 /10万 ,大约 4 0 %病例发生在 1周岁以内 ,10 %发生…     

接种b型流感嗜血杆菌疫苗致重症荨麻疹1例

患儿女，9个月，因接种b型流感嗜血杆菌（Hib）疫苗4小时全身出现风团样皮疹，于2006年6月12日2pm就诊。4小时前患儿接种Hib疫苗（上海葛兰索史克生物制品有限公司生产，批号为B342BA，商品名为贺新立适，有效期2008年8月），4小时后注射部位（左上臂三角肌附着处）出现红肿，即而全身瘙痄，随后出现风团样皮疹，不伴有发热，无咳嗽、气喘，无腹痛及腹泻。查体：T37℃，P108次／分，R29次／分，WT8Kg。急性病容，     

接种b型流感嗜血杆菌结合疫苗有讲究

并非所有儿童都可以接种b型流感嗜血杆菌结合疫苗，必须讲究方式方法，否则会导致接种儿童发生不良反应，甚至危及生命。b型流感嗜血杆菌结合疫苗的接种对象是2月龄－5周岁的儿童。接种疫苗后，可刺激机体产生抗b型流感嗜血杆菌的免疫力，用于预防由b型流感嗜血杆菌引起的脑膜炎、肺炎、败血症、蜂窝组织炎、关节炎等感染性疾病。     

不可分型流感嗜血杆菌重组Hap蛋白的表达及其生物学活性分析

目的 在原核系统中表达并纯化不可分型流感嗜血杆菌(NTHi)的重要粘附因子Hap蛋白,并对其免疫原性和粘附活性进行初步研究.方法 IPTG诱导重组质粒pET32a(+)-Hap在E.coli BL21中高效表达,Ni2+亲和层析柱纯化表达产物.用纯化的Hap重组蛋白进行体外竞争黏附实验,SEM观察及菌落形成单位计数法分析该重组蛋白的黏附活性.纯化蛋白与黏膜免疫佐剂CT-B联合鼻腔免疫BALB/C小鼠,ELISA检测小鼠抗Hap的IgA及IgG抗体水平.结果 纯化产物的SDS-PAGE分析显示获得单一的目的 蛋白条带,Gel analysis软件分析蛋白纯度可达85%,纯化产物超滤浓缩后,测得其蛋白浓度为3.2 g/L.体外竞争黏附实验观察到Hap重组蛋白的加入可明显抑制NTHi对胞外基质的黏附,且细菌黏附量的减少与对照组相比差异具有统计学意义(P<0.01).该重组蛋白与免疫佐剂CT-B联合免疫小鼠.可刺激机体产生较高水平的IgG或IgA抗体,与重组抗原单独免疫组相比,差异具有显著性(P<0.05).结论 Hap蛋白在原核系统中获得较高浓度和纯度的表达,纯化Hap重组蛋白具有良好的免疫原性和粘附活性.     

Ampicillin resistance in Haemophilus influenzae

Fourteen out of 150 (9.3%) consecutive strains of Haemophilus influenzae isolated on culture of sputum in the Bacteriology Department, Belfast City Hospital, during 1982/83 were found to be ampicillin-resistant (β-lactamase-producing). Susceptibility testing to other antibiotics of these ampicillin-resistant strains showed that cefuroxime, cefotaxime, gentamicin, and amoxycillin with clavulanic acid were reliable alternatives. Other useful alternatives included tetracycline, trimethoprim and co-trimoxazole. Erythromycin was of limited usefulness.     

Haemophilus influenzae meningitis in school-aged children

The relative frequency of meningitis caused by Haemophilus influenzae in school-age children was determined by reviewing etiologic diagnoses in children 6 to 15 years old admitted to four hospitals from 1974 to 1978. Sixty-five (45%) of 145 patients had aseptic meningitis and 29 (20%) had bacterial meningitis. Thirty-two (22%) of the patients had received antibiotic therapy before diagnosis, and 19 (13%) could not be classified. Six (21%) of the 29 patients with bacterial meningitis had H influenzae meningitis. Although aseptic disease was the most common type of meningitis, initial antibiotic therapy for presumed bacterial meningitis in school-aged children should include adequate coverage for H influenzae.     

Haemophilus influenzae pneumonia in adults.

Thirty cases of Haemophilus influenzae pneumonia with clinical and laboratory features have previously been recorded in adults. During the past three years, we have examined 18 patients in whom this diagnosis was established by transtracheal aspirate or blood culture. Our study suggests that H influenzae, both typable and nontypable strains, is a more frequent cause of pneumonia in adults than previously appreciated. We found no clinical values that distinguished H influenzae pneumonia from other bacterial pneumonias. A properly performed Gram's stain of a transtracheal aspirate specimen is classical in its appearance and facilitates instritution of appropriate initial treatment. The emergence of both typable and nontypable organisms resistant to ampicillin makes it important that organisms be isolated from reliable samples for sensitivity testing. With appropriate therapy, the prognosis for patients with H influenzae pneumonia appears to be good.     

Secondary Haemophilus influenzae type b in day-care facilities. Risk factors and prevention

The risk factors for acquisition of secondary day-care-associated Haemophilus influenzae type b disease were evaluated in a cohort of children in Seattle-King County, Washington; Atlanta; and the state of Oklahoma. During the study period, 129 primary cases were identified in children less than 5 years old who attended day-care facilities. In ten instances (8%), a secondary case occurred between one and 60 days after a primary case in the same classroom. Risk of secondary disease in classroom contacts was strongly age related: 2.4% in children 0 to 11 months old, 1.2% in children 12 to 23 months old, and 0.0% in children 24 to 59 months old. Controlling for age, children attending day-care more hours per week were more likely to transmit or acquire secondary disease. Risk of secondary disease for children in other classrooms at a center where a case had occurred was not significantly greater than risk of primary disease. Administration of rifampin to classroom contacts of a child with invasive H influenzae was effective in preventing secondary cases (95% confidence interval for rifampin efficacy, 47% to 100%). For children 0 to 23 months old not treated with rifampin, risk of secondary disease was 2.7% (95% confidence interval, 1.1% to 4.3%), a risk approaching that reported in household contacts.