Vero细胞口服狂犬病疫苗的研究

本试验采用ＣＴＮ－１株经Ｖｅｒｏ细胞传１５代，病毒滴度均在７．０～８．０ｌｏｇＬＤ５０／ｍｌ之间。给８只犬分别口服８．１ｌｏｇＬＤ５０的疫苗３０天的中和抗体几何平均效价为１：１１０，中和抗体的平均国际单位为２．５９ＩＵ／ｍｌ，阳转率为１００％；给４只犬分别口服７．４３ｌｏｇＬＤ５０的疫苗，阳转率为５０％，中和抗体的平均国际单位为２．４８ＩＵ／ｍｌ。选用３５－６０日龄的乳犬１６只口服１０个剂量的疫苗，并在口服后５、１０、２０天、３个月各杀３只犬取脑及唾液腺分别在ＢＨＫ２１细胞上盲传２代，同时在昆明种小白鼠乳鼠脑内盲传２代，进行病毒分离，均为阴性，余下犬观察１２个月均正常。总之该口服狂犬病疫苗对犬具有较好的安全性及口服免疫原性。     

Rabies control in Japan

In 1957 Japan succeeded in eradicating rabies, which had been endemic since the 18th century, due to the registration and confinement of family dogs, the elimination of stray dogs, and the compulsory vaccination of dogs. At present, however, vaccination coverage of family dogs is far lower than the required level of 70%. The facilities that are presently able to investigate rabies are limited in number. In addition, few medical institutions keep rabies vaccine in stock or offer postexposure vaccination to travelers bitten by animals in rabies endemic areas. Moreover, rabies immunoglobulin (RIG) cannot be given to such individuals because RIG is not produced at present in Japan, nor is it authorized to be imported. To keep Japan free from any rabies deaths, an improvement in vaccination coverage among dogs and in the supply of postexposure prophylaxis is required, and the establishment of a rabies surveillance system is also considered to be essential.     

Feasibility and Effectiveness Studies with Oral Vaccination of Free-Roaming Dogs against Rabies in Thailand

(1) Background: Thailand has made significant progress in reducing the number of human and animal rabies cases. However, control and elimination of the last remaining pockets of dog-mediated rabies have shown to be burdensome, predominantly as a result of the large numbers of free-roaming dogs without an owner that cannot be restrained without special efforts and therefore remain unvaccinated. To reach these dogs, the feasibility, and benefits of oral rabies vaccination (ORV) as a complementary tool has been examined under field conditions. (2) Methods: ORV of dogs was tested in five study areas of four provinces in Thailand. In these areas, sites with free-roaming dogs were identified with the support of local municipal workers and dog caretakers. ORV teams visited each of five study areas and distributed rabies vaccine (SPBN GASGAS) in three bait formats that were offered to the dogs using a hand-out and retrieval model. The three bait types tested included: egg-flavored baits, egg-flavored baits pasted with commercially available cat liquid snack, and boiled-intestine baits. A dog offered a vaccine bait was considered vaccinated when the discarded sachet was perforated or if a dog chewed vaccine bait at least 5 times before it swallowed the bait, including the sachet. (3) Results: A total of 2444 free-roaming dogs considered inaccessible for parenteral vaccination were identified at 338 sites. As not all dogs were approachable, 79.0% were offered a bait; of these dogs, 91.6% accepted the bait and subsequently 83.0% were considered successfully vaccinated. (4) Conclusion: Overall, 65.6% of the free-roaming dogs at these sites were successfully vaccinated by the oral route. Such a significant increase of the vaccination coverage of the free-roaming dog population could interrupt the rabies transmission cycle and offers a unique opportunity to reach the goal to eliminate dog-mediated human rabies in Thailand by 2030.     

狂犬病基因工程灭活疫苗（HEP-Flury-dG株）的免疫效力

目的：为了检测狂犬病基因工程灭活疫苗（HEP-Flury-dG株）注射犬只后的免疫效力。方法：在广州市增城区某村进行狂犬病基因工程灭活疫苗（HEP-Flury-dG株）环境释放试验,登记该村所有家养犬的信息并存档,同时进行疫苗免疫。随机采集免疫前血清样品66份,免疫后21d仍获得血清样品66份,使用ELISA方法检测样品的抗体水平。结果：免疫前该村犬只抗体阳转率为26%,免疫后21d抗体阳转率达到83%。经统计分析表明免疫前后抗体水平差异具有统计学意义;性别不是影响抗体水平变化的因素,但年龄可影响抗体水平变化,成犬抗体水平的增加明显高于幼犬。结论：狂犬病基因工程灭活疫苗（HEP-Flury-dG株）免疫犬只后可提供足够的保护力,有效预防狂犬病的传播;为农村地区家养犬建立免疫档案,是预防农村地区狂犬病发生的一种可行、有效的方法。     

Economical multi-site intradermal regimen with purified chick embryo cell vaccine (Rabipur) prevents rabies in people bitten by confirmed rabid animals.

OBJECTIVE: To determine the efficacy of a cost-effective multi-site intradermal regimen with purified chick embryo cell vaccine (PCECV, Rabipur) in preventing rabies in people bitten by confirmed rabid dogs. METHODS: Thirty-two people of different age groups who were severely bitten by confirmed rabid dogs were immunized with PCECV using the WHO recommended multi-site intradermal regimen of 0.1 mL of vaccine at eight sites on day 0, at four sites on day 7, and at one site each on days 28 and 90. In addition, passive immunization with human or equine rabies immunoglobulin was administered to 22 of these people before administering vaccine. They were followed for up to 3 years with periodic estimation of neutralizing antibody levels in their serum by mouse neutralization test (MNT). RESULTS: There was an excellent immune response with more than protective titers (>0.5 IU/mL) on all days tested up to the end of the 3-year observation period. More significantly, protective titers were seen in all subjects by day 7. Only minimal side effects were observed. All the patients were doing well at the end of the 3-year observation period, which is generally considered to be the maximum incubation period for rabies in humans. CONCLUSIONS: It can be concluded that this multi-site regimen with or without passive immunization has prevented the development of rabies encephalitis in these people bitten by confirmed rabid dogs. This should encourage more such studies, so that this cost-effective economical regimen with safe and potent cell culture vaccines can replace highly reactogenic neural tissue-derived Semple vaccine in developing countries such as India.     

狂犬病口服疫苗对消除人间狂犬病的重要作用北大核心CSCD

狂犬病严重威胁人类生命,控制狂犬病需要从病毒的源头着手。在狂犬病毒的多种宿主中,犬科动物所占比例最大。因此,消除狂犬病的关键是对犬的免疫覆盖率达到70%以上。事实证明,通过注射灭活疫苗对犬只形成免疫屏障是一件困难的事情。采用口服疫苗策略控制野生动物狂犬病的成功案例给大家提供了新的思路,因此世界卫生组织建议采用狂犬病口服疫苗对犬进行免疫,用以提高犬只的免疫覆盖率,从而达到2030年消除和控制狂犬病的目的。     

Rabies in Thailand: 1990

Of the more than 100,000 courses of postexposure rabies treatment given in Thailand annually, 95% consist of brain tissue-derived vaccine without immune globulin. Rabies tissue culture vaccines and immune globulins are expensive by the standards of developing countries. When they are given according to either of two proven intradermal postexposure schedules, significant savings can be achieved without loss of efficacy. Purified equine rabies immune globulins account for approximately 10% of the cost of human products administered to exposed individuals and have been shown to be safe and effective. A canine preexposure immunogenicity study with a potent, inactivated tissue culture vaccine revealed that 12.5% of Thai dogs failed to develop protective antibody titers 2 months after one subcutaneous injection. Previous studies have shown significant antigenic differences between Thai street rabies virus and European and North African strains.     

中国狂犬病口服疫苗的研究进展及其应用的紧迫性

中国科学家早在上世纪80年代开始研究狂犬病动物口服疫苗,获得数株毒力减弱的疫苗株,并进行过犬的口服免疫实验和大规模安全性和保护效果的现场观察。其中有几株减毒株的弱毒特征、遗传稳定性和免疫性与WHO认可并广泛应用的动物口服疫苗毒种相似。消除狂犬病的关键措施是对犬进行大面积免疫达到免疫覆盖率70%以上,中国犬的数量多达亿头,而且多数为散养犬,以常规的注射免疫很难达到这个水平。对犬实施口服免疫提高免疫率也是WHO和狂犬病专家积亟提倡和推荐的。中国已有多株减毒株,国家应充分发挥它们的作用,在人力、物力、财政上予以大力支持,完善这些候选株以便尽快得到批准应用,从而达到全球2030年消除人间狂犬病的目标。     

Review of Oral Rabies Vaccination of Dogs and Its Application in India

Oral rabies vaccines (ORVs) have been in use to successfully control rabies in wildlife since 1978 across Europe and the USA. This review focuses on the potential and need for the use of ORVs in free-roaming dogs to control dog-transmitted rabies in India. Iterative work to improve ORVs over the past four decades has resulted in vaccines that have high safety profiles whilst generating a consistent protective immune response to the rabies virus. The available evidence for safety and efficacy of modern ORVs in dogs and the broad and outspoken support from prominent global public health institutions for their use provides confidence to national authorities considering their use in rabies-endemic regions. India is estimated to have the largest rabies burden of any country and, whilst considerable progress has been made to increase access to human rabies prophylaxis, examples of high-output mass dog vaccination campaigns to eliminate the virus at the source remain limited. Efficiently accessing a large proportion of the dog population through parenteral methods is a considerable challenge due to the large, evasive stray dog population in many settings. Existing parenteral approaches require large skilled dog-catching teams to reach these dogs, which present financial, operational and logistical limitations to achieve 70% dog vaccination coverage in urban settings in a short duration. ORV presents the potential to accelerate the development of approaches to eliminate rabies across large areas of the South Asia region. Here we review the use of ORVs in wildlife and dogs, with specific consideration of the India setting. We also present the results of a risk analysis for a hypothetical campaign using ORV for the vaccination of dogs in an Indian state.     

中山市狂犬病105例回顾性分析

目的收集105例狂犬病完整的流行病学个案资料,分析流行特征,为本市狂犬病的预防控制提供参考。方法病例个案资料录入Epidata数据库,并用spss11.0统计软件包进行分析。结果发病季节以7-12月份为高峰;男性明显多于女性,其发病比例为2.28∶1;职业以农民为主,占55.24%;本市人狂犬病病例传染源以家犬为主,占93.33%;暴露后伤口未作任何处理和接种疫苗的占63.81%,自行处理或不规范处理伤口和无及时接种疫苗的占20.00%,及时处理伤口并注射狂犬疫苗但发病的仅占病例数的7.62%。暴露后潜伏期最短13d,最长1153d,平均潜伏期78.56d。结论建议进一步加强犬类的"管、免、灭";加强宣传和健康教育,使人们在被带狂犬病毒的犬类咬伤后及时获得狂犬病暴露后的免疫预防;尽可能地对高危人群和高发地区开展暴露前免疫,降低狂犬病发病率。     

A study on the seroconversion in dogs vaccinated with cell culture rabies vaccine

Seroconversion studies were conducted in dogs vaccinated with rabies vaccine, of different age group, sex and breed, with a single and booster doses of cell culture Rabies vaccine. The results of the present study revealed that (i) Maternal antibodies were detected in 40% to 80% of 60 selected pet dogs. However 20% to 60% of the pet dogs did not have protective levels of antibody (<0.5IU). (ii) A single dose of vaccine resulted in appreciable levels of protective antibody in 100% of pet dogs both in seronegetive and in dogs with low levels of antibody. (iii) When a booster dose was administered at twelve months period, considerable levels of antibody persisted upto twenty-four months. It can be concluded from the present study that a single dose of potent tissue culture vaccine resulted protective levels of antibody in the seronegetive dogs (<0.5IU). In an Rabies endemic country like INDIA, annual booster dose of vaccine would enhance the immune response and help in the persistence of protective levels of antibody in the immunized dogs.     

Facial dog bites in Southwestern Nigerian children: an analysis of eight cases

This paper analyses the prevalence, pattern and treatment outcome of eight cases of facial dog bite in Nigerian children seen and managed at the Obafemi Awolowo University Teaching Hospital's Complex, a tertiary hospital located in Southwestern Nigeria. During the study period of 18 months (July 1996-December 1997), a total of 174 cases of facial soft tissue injuries in children were managed, two of which resulted in rabies and death of the patients. Based on the findings of this analysis, we suggest that unprovoked facial dog bites inflicted by familiar domestic dogs may be complicated by rabies. Ignorance about the risk of uncompleted treatment, financial constraints and scarcity of the vaccine may be implicated in the outcome. The need to improve community knowledge of rabies and the availability and affordability of rabies vaccine are highlighted.     

云南省狂犬病病人和犬类感染狂犬病病毒及M基因序列分析

目的 调查云南省部分地区狂犬病病人和疫点犬群携带狂犬病病毒(rabies virus, RABV)状况及RABV膜基质蛋白(matrixprotein, M)基因序列分析,为狂犬病防控提供科学依据。方法 2008-2009年在云南省采集犬脑组织标本606份,狂犬病病人唾液8份,脑脊液1份,用直接免疫荧光试验检测RABV抗原,用RT-PCR检测RABV核酸,对阳性标本进行M基因序列测定和分析。结果 所有标本经检测,RABV抗原和/或核酸阳性16份,其中疫点扑杀的貌似健康犬脑组织3.10%(10/323),狂犬病病人唾液5份、脑脊液1份。狗肉餐馆屠宰犬的脑组织283份均为阴性。云南16株RABV的M基因核苷酸和氨基酸同源性分别为88.5%~100%和85.2%~99.5%。它们与中国人用疫苗株aG核苷酸和氨基酸同源性分别为83.9%~85.7%和82.3%~93.6%;与人用疫苗株CTN181核苷酸和氨基酸同源性分别为99%~99.7%和98.5%~99%。系统进化分析表明,云南16株RABV均属基因Ⅰ型并可分为进化Ⅰ和Ⅱ群并分别与泰国等东南亚国家和相邻省份流行株具有较近的亲缘关系。结论 云南省狂犬病流行与周边省份和东南亚地区的狂犬病传播扩散有一定关系;狂犬病疫点部分貌似健康犬携带RABV并具有传染源意义;云南狂犬病病毒株M基因与我国人用狂犬病疫苗CTN株的同源性和亲缘关系较近,但与aG株同源性存在。     

西安市6 635例狂犬病暴露者流行病学调查

目的 分析西安市2016-2017年来6 635例狂犬病暴露者流行特征,探讨相关危险因素,为狂犬病的防治提供依据。方法 对狂犬病暴露者按年龄、性别、职业、暴露时间、暴露地点、暴露动物、暴露部位、暴露级别和处置及发病情况等进行统计分析。结果 6-10月为狂犬病暴露高发季节,共暴露4 038例,占60.86%;犬和猫为主要致伤动物,占66.25%和30.49%;暴露男女比为1∶1.19;20~岁及10~岁年龄组暴露最多,占33.56%和16.38%;暴露者以自由职业者、学生及服务业从业者居多,分别占总26.36%、21.75%和15.33%;暴露地点主要在家中,占67.94%;暴露部位以上、下肢为主,分别占43.48%和38.63%;以III级暴露为主,占80.24%;暴露后规范处置率达100%,狂犬病疫苗接种率达100%,疫苗全程接种率为99.83%,III级暴露后被动免疫制剂注射率为52.95%。结论 西安市面临着严峻的狂犬病防控压力,其狂犬病暴露流行病学特征与周边地区及陕西省全省不尽相同,有自己的特点,应重点加强宠物犬猫的免疫与管理,消灭流浪犬猫;针对不同人群特点加强狂犬病预防及处置知识的健康教育,提高其预防意识和认知水平;进一步加强犬伤处置门诊规范化建设。     

The epidemiology of animal bite injuries in Uganda and projections of the burden of rabies

OBJECTIVES: Rabies is a global problem, although it is often under-reported in developing countries. We aimed at describing the profile of patients presenting to health centres with animal bite injuries in Uganda, and use a predictive model to estimate the mortality of rabies at a national level. METHODS: We conducted a passive surveillance study in Uganda based in a random sample of health centres supplied with rabies vaccine to determine the characteristics of bite injury patients and establish the age and sex profiles of patients, the site of bites and their severity, wound management techniques and details of the vaccination course given. We also applied a decision tree model to the data to estimate the rabies mortality from the bite injury data using an established protocol. RESULTS: We found that most patients are bitten by dogs, and that a considerable proportion of these are young children, who are at greater risk of developing rabies in the absence of treatment due to the location of the bites they receive. From conservative parameter estimates, we estimate that in the absence of post-exposure prophylaxis (PET), 592 (95% CI 345-920) deaths would occur, and that if one dose of PET is sufficient for protection following a rabid animal bite, 20 (95% CI 5-50) deaths would occur annually. If a complete course of PET is required for protection following a rabid animal bite, up to 210 (95% CI 115-359) deaths would occur, as 41% of patients did not complete their course of PET. CONCLUSIONS: Active animal bite surveillance studies are required to improve our mortality estimates and determine the true burden of rabies in the Ugandan population. We emphasize the need for small-scale active case detection studies and improved data on the recognition of rabies in dogs as inputs for improving national-level estimates of rabies mortality.     

Equine rabies immune globulin: a product with an undeserved poor reputation

Four hundred nineteen patients exposed to rabies in Thailand were treated with equine rabies immune globulin (ERIG) manufactured by Sclavo of Italy, a product also licensed in the United States of America. They were followed for a minimum of 1 month after ERIG injection and rabies vaccine administration. Adverse serum sickness-like reactions were noted in 15 patients (3.58%). These were clinically acceptable and only 1 of these patients required corticosteroid therapy and short term hospitalization for serum sickness. ERIG is approximately 1/10 of the cost of human rabies immune globulin (HRIG), which is not generally available in developing countries. ERIG is a safe and underutilized essential biological when HRIG is not affordable or available.     

Current rabies vaccines and prophylaxis schedules: preventing rabies before and after exposure

Travellers are probably the largest group in the general population to receive rabies pre-exposure prophylaxis. The dangerous consequences of the unavailability of rabies immune globulin in many countries could be ameliorated if pre-exposure rabies vaccination were practised more widely, especially in children, living in dog rabies enzootic countries. The WHO has recommended several different regimens for post-exposure prophylaxis, while individual countries decide on protocols for local use. Intramuscular regimens are expensive and waste vaccine. Although failure to receive vaccine is usually the due to the cost, the economical potential of intradermal vaccination has still not been realised 19 years after its introduction. The currently recommended 2-site intradermal post-exposure regimen is not economical for use in rural areas where 80% of Indian rabies deaths occur. Most countries using it demand higher potency vaccine, indicating that they do not have complete confidence in the method. This intradermal regimen has only been used where immunoglobulin is likely to be available for severely bitten patients. Increased intradermal doses are sometimes used for selected patients. Provision of economical rabies prophylaxis can be improved. Decisions to change recommendations should take account of the immunological, financial, practical and logistical aspects of dog bite treatment in remote areas.     

Comparative Study of Optical Markers to Assess Bait System Efficiency Concerning Vaccine Release in the Oral Cavity of Dogs

Oral vaccination of dogs against rabies has the potential to achieve mass coverage and thus deplete the virus of its most important reservoir host species. There is, however, no established non-invasive method to evaluate vaccine release in the oral cavity, following bait ingestion. In this study, two pre-selected marker methods in conjunction with their acceptance were assessed in local Thai dogs. Shelter dogs (n = 47) were offered one of four randomized bait formulations; bait type A-, containing Green S (E142) in a fructose solution; type B-, containing Patent Blue V (E131) in a fructose solution; type C-, containing the medium used for delivery of oral rabies vaccine in baits commercially produced; and type D-, containing denatonium benzoate, which was to serve as the negative control, due to its perceived bitterness. Patent Blue V was found to possess overall stronger dyeing capacities compared to Green S. Furthermore, there was no significant difference in the acceptance or bait handling of Patent Blue V baits compared to those containing the oral rabies vaccine medium alone, suggesting the potential use of this dye as a surrogate for rabies vaccine when testing newly developed bait formats.     

Epidemiologic trends of rabies in domestic animals in southern Thailand, 1994-2008

Rabies and associated risk factors in dogs, cats and cattle (n = 3,454) in southern Thailand during 1994-2008 were evaluated by using a mixed-effect logistic regression model. Overall prevalence was 48%. In dogs, odds of being rabid were 1.7 times higher in unvaccinated dogs than in vaccinated dogs and two times higher in dogs with bite history than in dogs with no known bite history. Similarly, aggressive dogs were more likely to be rabid than non-aggressive dogs. In cattle, aggression, pharyngeal paralysis, hyperactivity, and depression were clinical signs associated with being rabid. Annual fluctuations of the species-specific prevalence of rabies is suggestive of a positive correlation between canine and either feline (r = 0.60, P = 0.05) or bovine rabies (r = 0.78, P = 0.004). Insufficient vaccination coverage led to maintenance of rabies, which could be easily controlled by increased vaccine coverage and public education.     

Effective preexposure and postexposure prophylaxis of rabies with a highly attenuated recombinant rabies virus

Rabies remains an important public health problem with more than 95% of all human rabies cases caused by exposure to rabid dogs in areas where effective, inexpensive vaccines are unavailable. Because of their ability to induce strong innate and adaptive immune responses capable of clearing the infection from the CNS after a single immunization, live-attenuated rabies virus (RV) vaccines could be particularly useful not only for the global eradication of canine rabies but also for late-stage rabies postexposure prophylaxis of humans. To overcome concerns regarding the safety of live-attenuated RV vaccines, we developed the highly attenuated triple RV G variant, SPBAANGAS-GAS-GAS. In contrast to most attenuated recombinant RVs generated thus far, SPBAANGAS-GAS-GAS is completely nonpathogenic after intracranial infection of mice that are either developmentally immunocompromised (e.g., 5-day-old mice) or have inherited deficits in immune function (e.g., antibody production or type I IFN signaling), as well as normal adult animals. In addition, SPBAANGAS-GAS-GAS induces immune mechanisms capable of containing a CNS infection with pathogenic RV, thereby preventing lethal rabies encephalopathy. The lack of pathogenicity together with excellent immunogenicity and the capacity to deliver immune effectors to CNS tissues makes SPBAANGAS-GAS-GAS a promising vaccine candidate for both the preexposure and postexposure prophylaxis of rabies.