毒种感染剂量及小牛血清浓度对腮腺炎减毒活疫苗生产的影响

发布时间：2018-08-22 09:34

【摘要】：流行性腮腺炎由腮腺炎病毒引起,为影响儿童及青少年的急性传染病。除侵犯腮腺外,可引起脑膜炎、脑膜脑炎、睾丸炎、卵巢炎、胰腺炎、乳腺炎等,可与腮腺炎同时发生,也可发生于腮腺炎之前或之后,或者单独出现[11]。腮腺炎病毒属副粘病毒,系单股核糖核酸型。人是本病毒的唯一储存宿主。腮腺炎病毒可在鸡胚及组织培养如猴肾、人羊膜、HeLa细胞中繁殖。含有病毒颗粒抗原(V抗原)及可溶性抗原(S抗原)2种抗原成分。腮腺炎病毒很少变异,一般仅存在一种抗原型别。病毒颗粒具有血凝素,可与鸡等红细胞发生凝集反应,并可用以测定病人血凝抑制抗体。S抗原多用以进行补体结合试验。病毒在56℃经20分钟,稀乙醚经30分钟,1%来苏、乙醇、甲醇经2-5分钟皆可灭活。本病仅发生于人类,人群对本病普遍易感。传染源为早期患者和隐性感染者。发病前7天病毒已在唾液中出现,并持续至发病后9天。表现为睾丸炎或脑膜炎而无腮腺炎的患者,也有病毒自唾液排出。[2]根据血清学及流行病学调查,大流行中隐性感染占30-40%,并有病毒自唾液中排出,成为重要的传染源。本病借唾液飞沫传播,全年皆有病例发生,多数发生于晚冬及初春。15岁以下儿童及青少年约占发病人数90%。成人80%可测出抗体。一次感染后可有持久免疫力,很少再次感染。腮腺炎病毒进入人体后的发病过程,存在二种假说。一种认为病毒由口经腮腺导管如睾丸、脑、胰腺等中致病。另一种认为病毒最初在呼吸道表层上皮中复制,以后进入血中随循环播散至腮腺及其他器官并在其中增殖,再自这些器官第二次进入血中,侵犯第一次病毒血症未侵及的器官。有些病例仅有其他器官受侵而无腮腺受累,或其他器官先于腮腺受累的事实,似更支持第二种假说。腮腺的非化脓性炎症是本病的主要病理改变。腮腺中有浆液纤维性渗出,腮腺导管周围及腺体壁有淋巴细胞浸润。导管可阻塞或扩张。睾丸受累时轻者仅有间质水肿及浆液纤维性渗出,重者可有淋巴细胞浸润、曲精小管上皮破坏及堵塞。腮腺炎脑膜炎病变轻微,尸检材料很少。脑炎病变主要为急性血管周围脱髓鞘改变,与感染后脑炎无大差别。潜伏期14-21天。前驱期症状有乏力、食欲不振等,继之腮腺肿大作痛,进酸性食物后痛尤明显。腮腺肿大可限于一侧,但多数病人一侧肿大后1-4天又累及对侧。一般以耳垂为中心,向前、后、下发展,状如梨形,具坚韧感,边缘不清。局部皮肤紧张发亮,表面灼热,但多不红,有轻触痛。颔下腺或舌下腺可同时或少数情况下单独受累。腮腺肿大于2-3日达高峰,持续4-5日后消退。病人发热38℃左右,持续约一周。一般成人患者症状较儿童为重。睾丸炎多见于成人,多发生于肿大的腮腺开始消退之时。病人又发高热,睾丸肿痛。睾丸多数为单侧,全病程10日左右。虽然1/3病人有不同程度的睾丸萎缩,但即或双侧受累,也很少引起不育。腮腺炎病毒极易侵犯中枢神经系统,可引起脑膜炎、脑膜脑炎。脑脊液检查至少一半以上的腮腺炎病人有改变。临床有头痛、颈强直表现者,约占腮腺炎病例的10%。全病程约10日,大多恢复而无后遗症。腮腺炎病毒尚可引起乳腺炎、卵巢炎、胰腺炎、心肌炎、甲状腺炎、肾炎、多关节炎以及单侧神经性耳聋等。血白细胞计数正常或减低。有睾丸炎者白细胞总数可增多。腮腺炎患者血淀粉酶增高。有些脑膜炎病例,临床上虽无腮腺炎,但血淀粉酶增高,可能为亚临床型腮腺炎。有胰腺炎的病例除血淀粉酶增高外,血脂肪酶亦高。脑膜炎病例脑脊液细胞数增加,以淋巴细胞为主,蛋白正常或升高。脑膜炎病例脑脊液细胞数增加,以淋巴细胞为主,蛋白正常或升高,糖量正常,但10-20%病例亦可减低。典型腮腺炎病例不难诊断。本病应与化脓性腮腺炎、腮腺肿瘤、药物及其他病毒(如副流感病毒、柯萨奇甲组病毒)引起的腮腺肿大鉴别。确诊(包括无临床腮腺炎的脑膜炎或睾丸炎病人)有赖病毒分离及血清学检查。病毒分离可取血、咽漱液、腮腺导管分泌物、脑脊液及尿,接种组织培养细胞或鸡胚羊膜腔。血清学检查常用的方法为补体结合试验。S抗体出现早,下降较快,6-12月后一般已不能测出。V抗体出现晚,迟于S抗体1-2周,效价常较S抗体为高,且持续多年。急性期及恢复期双份血清S及V抗体效价4倍增加时可以确定诊断。S抗体高而无V抗体者可初步认为是新病例的早期感染;有V抗体而无S抗体者可认为过去曾受感染。还可测定中和抗体及血凝抑制抗体。本病无特异治疗,主要对症及支持治疗。重型睾丸炎及脑膜炎患者可用肾上腺皮质激素,可减轻症状,但不能防止睾丸萎缩。腮腺炎、脑膜炎、睾丸炎预后好很少复发或死亡。腮腺炎病人应隔离至临床症状消失为止。丙种球蛋白一般无被动免疫效果。对1岁以上儿童及青少年未患过腮腺炎者,可用腮腺炎减毒活疫苗免疫注射。也有将腮腺炎病毒与麻疹及风疹病毒联合制备疫苗。接种95%血中抗体阳性、腮腺炎发病率降低,抗体持续最少5年。现在做为预防腮腺炎病毒的最好方法就是注射腮腺炎减毒活疫苗,而现有的生产工艺为以SPF鸡胚细胞做为腮腺炎病毒感染的基础细胞,然后在适宜温度下培养,最后收获病毒液[3]。因此,在此生产工艺中毒种的感剂量及小牛血清浓度将是两个非常重要的因素,找出最佳的毒种剂量及合适的血清浓度会对腮腺炎减毒活疫苗的产量及质量有明显的提高。
[Abstract]:Epidemic mumps are caused by mumps virus and are acute infectious diseases affecting children and adolescents. In addition to invading the mumps, it can cause meningitis, meningoencephalitis, orchitis, ovaritis, pancreatitis, mastitis, etc. It can occur simultaneously with mumps, before or after mumps, or alone [11].
Mumps virus is a paramyxovirus, a single stranded ribonucleic acid type. It is the only storage host of the virus. Mumps virus can reproduce in chicken embryos and tissue cultures such as monkey kidney, human amniotic membrane, and HeLa cells. It contains two antigenic components: virus granule antigen (V antigen) and soluble antigen (S antigen). Mumps virus is rarely mutated and generally only exists. Virus particles have hemagglutinin, can agglutinate with chicken and other red blood cells, and can be used to detect hemagglutination inhibitory antibodies in patients. S antigen is often used for complement binding tests. Viruses can be inactivated by dilute ether for 30 minutes, 1% Lesu, ethanol and methanol for 2-5 minutes at 56 C for 20 minutes.
The disease occurs only in humans and is generally susceptible to the disease in the population. Infectious sources are early patients and latent infections. Viruses appear in saliva seven days before onset and persist until nine days after onset. The disease is transmitted by salivary droplets throughout the year. Most cases occur in late winter and early spring. Children and adolescents under 15 years old account for about 90% of the cases. 80% of adults can detect antibodies. Once infected, the disease has a lasting immunity and rarely re-infection.
There are two hypotheses about the pathogenesis of mumps virus when it enters the human body. One hypothesis is that the virus is caused by oral passage through parotid ducts such as testis, brain, pancreas, etc. The other hypothesis is that the virus first replicates in the surface epithelium of the respiratory tract, then enters the blood and circulates to the parotid gland and other organs and multiplies therein, and then from these organs to the second. Non-suppurative inflammation of the parotid gland is the main pathological change of the disease. Serous fibrillary exudation occurs in the parotid gland and the periductal area of the parotid gland. Lymphocyte infiltration and lymphocyte infiltration were found in the wall of the gland. The duct could be obstructed or dilated. Leydig edema and serous fibrillary exudation were found only in the mild cases. Lymphocyte infiltration was found in the severe cases. The epithelium of seminiferous tubules was destroyed and blocked. The incubation period is 14-21 days. The prodromal symptoms are fatigue and loss of appetite, followed by swelling and pain of the parotid gland, especially after eating acidic food. The parotid gland swelling reaches its peak in 2-3 days and subsides after 4-5 days. The patient has a fever of about 38 degrees centigrade, lasting about a week. The symptoms of adult patients are more serious than those of children. Orchitis is more common in adults and more common in adults. When the swollen parotid gland begins to subside.
Most of the testicles are unilateral, and the course of the disease is about 10 days. Although one third of the patients have varying degrees of testicular atrophy, or bilateral involvement, but also rarely cause infertility. Mumps virus is very easy to invade the central nervous system, can cause meningitis, meningoencephalitis. Cerebrospinal fluid examination at least half of the patients with mumps have changed. Mumps virus can also cause mastitis, ovaritis, pancreatitis, myocarditis, thyroiditis, glomerulonephritis, polyarthritis and unilateral nervous deafness. Blood white blood cell count is normal or reduced. People with orchitis are white and thin. In some cases of meningitis, although there is no mumps, the serum amylase may be increased, which may be subclinical type of mumps. In some cases of pancreatitis, besides the increase of serum amylase, the serum lipase is also high. In meningitis, the number of cerebrospinal fluid cells is increased, mainly lymphocytes, and the protein is normal or rising. The number of cerebrospinal fluid cells in meningitis patients increased, mainly lymphocytes, normal or elevated protein, normal sugar content, but 10-20% of cases can also be reduced. Typical cases of mumps are not difficult to diagnose. This disease should be differentiated from parotid gland enlargement caused by suppurative mumps, parotid tumors, drugs and other viruses (such as parainfluenza virus, Coxsackie A virus). Confirmed diagnosis (including meningitis or orchitis without clinical mumps) depends on virus isolation and serological examination. Virus isolation can take blood, pharyngeal gargle, parotid duct secretion, cerebrospinal fluid and urine, inoculate tissue-cultured cells or chicken embryo amniotic cavity. Serological tests commonly used are complement-binding tests. S antibodies appear early, decline faster, 6-12 After the month,.V antibody was not detected late, but later than S antibody for 1-2 weeks.
It is usually higher than S antibody and lasts for many years. Diagnosis can be made when the titers of S and V antibodies in both sera are increased four times in acute and convalescent stages. Severe orchitis and meningitis patients can use adrenocorticosteroids, can alleviate symptoms, but can not prevent testicular atrophy. Mumps, meningitis, orchitis prognosis is good rarely recurrence or death. Mumps should be isolated until the clinical symptoms disappear. Gamma globulin generally has no passive immune effect. For children and adolescents over one year old who have not suffered from mumps, live attenuated mumps vaccine can be used to immunize. There are also mumps virus and measles and rubella virus vaccine preparation. 95% of the blood antibody positive, mumps incidence decreased, antibody lasted for at least 5 years. Now the best way to prevent mumps virus is injection. Mumps live attenuated vaccine, and the existing production process for SPF chicken embryo cells as the basis of mumps virus infection cells, and then cultured at appropriate temperatures, and finally harvested the virus solution [3]. And the appropriate serum concentration will significantly improve the production and quality of live attenuated mumps vaccine.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

【参考文献】