珠海市动物从业人员SARS—CoV抗体血清流行病学研究

目的通过对动物从业人员以及其它健康人群感染SARS-CoV的血清学研究,揭示人群中SARS-CoV抗体水平,探讨动物作为传染源的传播途径.方法随机选取无SARS临床症状的健康动物从业人员以及其它健康人员等545人作为调查对象,使用统一调查表对调查对象进行个案调查,同时采集调查对象的血清,用ELISA法检测SARS-CoV抗体.结果共检测各种人群血样545份,其中动物从业人员血样269份,SARS-CoV抗体阳性率为8.18%;健康人群血样276份,阳性率为1.09%,动物从业人员SARS-CoV抗体阳性率和非从业人员SARS-CoV抗体阳性率比较有显著性差异.结论动物从业人员SARS-CoV抗体阳性率显著高于非从业人员,支持SARS-CoV源于野生动物的假设.     

广州市动物市场从业人员SARS冠状病毒感染的危险因素分析

目的 探讨动物市场从业人员SARS冠状病毒(CoV)感染的可疑危险因素。方法 在对广州市3家大型野生动物市场从业人员进行SARS-CoV抗体检测的基础上,采用自行设计调查表进行SARS-CoV感染的危险因素调查,采用单因素和多因素logistic回归分析方法分析资料。结果经单因素logistic回归分析发现,经营其他畜类(猫、狗)、野生畜类(狸、山猪、黄猄)、其他野生类(蝙蝠、巨蜥、穿山甲等)、水产类(甲鱼、龟、鳗鱼、蛙等)动物、采购销售等是SARS-CoV感染的相关因素;经多因素logistic回归分析发现有意义的因素是经营野生畜类(狸、山猪、黄猄)(P<0.001)和经营家禽(鸡、鸭、鹅、鸽子)(P=0.04),OR值分别为12.28、0.41。结论 接触狸类动物是该从业人员感染SARS-CoV的主要危险因素。     

应用RT-PCR方法检测人与动物的SARS冠状病毒

目的选择合适的RTPCR检测方法应用于人和与人类生活密切相关的多种野生、圈养和市售动物携带SARS样冠状病毒的检测,探索SARS样病毒的来源。方法收集深圳SARS临床诊断病例、疑似及观察病例咽漱液标本共350份,分别用TaqMan和分子信标荧光RTPCR法进行检测;收集386只动物的咽肛拭子样本及病毒分离培养物共442份样品进行了TaqMan荧光RTPCR和普通RTPCR法的检测。结果41例临床确诊病例荧光RTPCR法检出10份阳性,阳性率24.39%,TaqMan法和分子信标法检出结果8份符合(符合率88.89%)。对动物样本病毒分离培养物细胞病变样本18份进行检测,检出16份阳性,其中14份(87.5%)样本来源为果子狸。深圳东门市场及广东周边地区市场及养殖场动物样本检测结果:果子狸、貉及獾类动物总的阳性率为39.02%,而其它动物的阳性率为0,两者间差异具有显著性(P<0.01);对东门市场109只主要野生动物咽肛拭子样本进行PCR检测,阳性率44.04%,而145只野外的野生动物阳性率仅为0(P<0.01)。结论TaqMan法和分子信标法均可用于SARS检测;动物中尤其是果子狸等野生动物可能携带有SARS冠状病毒。     

惠州市人群SARS冠状病毒抗体血清学调查分析

目的分析惠州市各类人群SARS-CoV抗体水平情况。方法抽取惠州市某门诊健康体检人群、医务工作人员、学生、学龄前儿童、野生动物从业人员血清标本共2397人份,采用酶联免疫方法进行SARS病毒IgG抗体检测。结果2397份样本,SARS病毒IgG抗体阳性率为1.1%(26/2397),0～6岁阳性率为4.0%(20/501)。7～14岁、医务工作人员阳性率分别为0.95%(5/527),0.32%(1/311)。健康体检人群样本1028人份,其中厨师样本54人份,野生动物从业人员样本30人份,抗体均为阴性。结论惠州地区人群SARS-CoV抗体水平低下,不能形成免疫屏障,人群普遍易感,作为预警信号,提示应当适时作好相关预防工作。     

广州市2004年某野生动物市场动物携带SARS-CoV监测

目的　了解广州市某野生动物市场动物携带严重急性呼吸综合征冠状病毒 (SARS CoV)的动态变化 ,预测 2 0 0 4 - 2 0 0 5年冬季当地发生动物源性SARS的危险性。方法　在SARS发病相关月份采集广州市某野生动物市场动物的肛拭子和咽拭子标本 ,使用实时荧光聚合酶链反应 (PCR)初筛 ,使用RT PCR和序列分析方法核实。结果　在 2 0 0 4年 1月 5日广东省开始清除野生动物行动前采集的 31只动物 (5只赤狐、2 0只猫、6只黄毛鼠 )中 ,有 8只动物SARS CoV检测阳性 (2 5 .8% ) ,包括 3只赤狐、4只猫、1只黄毛鼠。在 2 0 0 4年 1月 2 0日采集的 119只动物 (6只兔、13只猫、4 6只原鸡、13只斑嘴鸭、10只灰雁、31只鹧鸪 )中 ,仅 1只灰雁的肛拭子检测阳性。 2 - 11月份采集的 10 2只动物 (14只灰雁、3只猫、5只兔、9只斑嘴鸭、2只鹧鸪、8只雉鸡、6只鸽、9只黄麂、19只山猪、16只黄毛鼠、5只犬、1只水貂、3只山羊、2只绿孔雀 )中 ,仅 4月份采集的 1只山猪的肛拭子标本阳性。 11- 12月份在广东省采集的 2 2只果子狸标本均为阴性。结论　和SARS流行或出现散发病例 2 0 0 3年 5月、2 0 0 4年 1月5日某野生动物市场动物的SARS CoV的RT PCR检测情况相比 ,2 0 0 4年 1月 2 0日至 12月广州市某野生动物市场的动物携带SARS CoV的比     

SARS疫苗的研究现状和挑战

严重急性呼吸综合征（Severe acute respiratory syndrome，SARS）是由SARS冠状病毒（SARS-CoV）引起的一种新发传染病。1997年在该病我国香港首先发现，2002年末在广东省再次出现，并沿国际航空线路迅速传至世界各地，导致全球五大洲33个国家8450人发病，810人死亡。SARS全球暴发严重威胁着全球公共卫生和社会经济的稳定，尽管这次暴发于2003年被控制，但由于实验室SARS—CoV的泄露事故而相继在中国大陆和台湾、新加坡发生SARS暴发。2003年末和2004年初，广东报告与SAR-CoV感染动物接触者的新感染明显不同于2002～2003年的暴发。这些事件提示，SARS流行可在未来任何时间因实验室样本外泄或动物宿主中由类SARS—CoV演化而来的SARS-CoV分离株导致再次暴发。     

广东省野生动物销售人员SARS冠状病毒抗体血清学研究

目的　比较野生动物销售人员和健康人群感染SARS冠状病毒的情况 ,为探索验证SARS冠状病毒源于动物的假设提供依据。方法　于 2 0 0 3年 5月选择广州市 3个野生动物市场销售人员、2家医院的医务人员、某疾病预防控制中心工作人员及某门诊健康体检人员等无SARS临床症状的健康人群为调查对象 ,采集调查对象的血清 ,用间接ELISA法检测SARS冠状病毒IgG抗体。结果　共检测销售野生动物人员血清 5 0 8份 ,SARS冠状病毒抗体阳性率为1 3 0 2 % (6 6 / 5 0 8) ,其中以主要销售果子狸人员的阳性率最高 ,为 72 7% (1 6 / 2 2 ) ,其次销售山猪人员为 5 7 1 % (1 6 / 2 8) ,销售黄犭京人员为 5 6 3% (9/ 1 6 ) ,销售兔人员为 4 6 2 % (6 / 1 3) ,销售山鸡人员为 33 3% (3/ 9) ,另在猫、鸟和蛇等其他 1 2种野生动物的销售人员中均不同程度检出SARS冠状病毒抗体。其他健康人群SARS冠状病毒抗体阳性率为 2 1 1 %(6 / 2 84 ) ,其中医院医务人员为 2 92 % (4 / 1 37) ,疾病控制人员为 1 5 9% (1 / 6 3) ,社区健康人员为 1 1 9% (1 / 84 )。销售野生动物人员SARS冠状病毒抗体阳性率与其他健康人群比较差异有显著性 (χ2 =2 6 0 89,P <0 0 1 )。结论　野生动物销售人员的SARS冠状病毒抗体阳性率显著高于其他健康人群     

广州市人群SARS血清IgG抗体检测结果分析

目的 了解广州市非典型肺炎人群血清中严重急性呼吸综合征(SARS)冠状病毒(CoV)IgG抗体的基本特征及有关影响因素。方法 采集恢复期SARS、非SARS肺炎患者及SARS患者家庭密切接触者、野生动物、蔬菜销售者、社区对照人群的血清标本,用ELISA方法检测血清中SARS-CoV IgG抗体,并对SARS患者进行流行病学调查。结果 上述人群中,临床诊断为SARS患者SARS-CoV IgG抗体检出率最高,为53.7％,野生动物销售人员次之,为16.7％,社区普通人群为0.9％。在临床诊断的SARS患者中,可溯源病例抗体阳性率为90.4％,高于不可溯源病例的抗体阳性率为19.6％;且可溯源病例中低年龄组和高年龄组抗体阳性率低于中青年组,而不可溯源病例中不同年龄组患者抗体阳性率无差异;不同性别SARS患者的IgG抗体检出率也无差异;SARS抗体检出率呈逐月降低的趋势。结论 SARS临床诊断病例的血清SARS-CoV IgG抗体检出率只有53.7％,剔除患者年龄、使用激素、试剂盒等因素外,主要原因在于存在临床误诊的可能;实验室检测与流行病学调查相结合可明显提高SARS患者诊断的准确性;SARS-CoV存在隐性感染的可能,其在病毒传播过程中的作用有待进一步研究。     

ELISA检测广东省各地鼠形动物SARS特异性抗体的初步研究

目的　了解广东省各地鼠形动物血清SARS冠状病毒特异性IgG抗体水平 ,为探讨鼠形动物尤其是家鼠类媒介生物 ,与SARS流行是否存在相关性提供参考。方法　在广东省各地与SARS相关或被认为可能相关的场所诱捕鼠形动物 ,留取血清 ,用葡萄球菌A蛋白 酶联免疫吸附实验 (SPA ELISA)及常规ELISA检测SARS冠状病毒特异性IgG抗体水平。 结果　在广东省各地共获取鼠形动物 (包括褐家鼠、黄胸鼠、小家鼠及臭 )血清 193份 ,用SPA ELISA检测 ,7份特异性抗体阳性 ,均为褐家鼠或黄胸鼠 ,阳性率 3 .6% ;其中 13 9份褐家鼠、黄胸鼠血清用常规ELISA重复检测 ,检出阳性 18份 ,阳性率 12 .9% ;SPA ELISA检测阳性者中 5份用常规法复检 ,4份仍为阳性 ;所有阳性者均来自广州、佛山、深圳 3地。结论　在广东省各地捕获的鼠形动物中 ,血清SARS冠状病毒特异性IgG抗体阳性者均为褐家鼠或黄胸鼠 ,阳性者的地域分布似乎与广东省SARS流行情况存在一定的相关性 ;SPA ELISA法检测Rats类家鼠血清SARS冠状病毒特异性IgG的敏感性可能低于常规ELISA法 ,而检测阳性者与SARS冠状病毒接触或感染的相关程度还需进一步的研究证明。     

清远市健康人群SARS冠状病毒抗体血清流行病学研究

目的 了解清远地区健康人群不同年龄、职业的SARS冠状病毒(简称SARS-CoV)抗体水平状况。通过对SARS流行后期(非疫区)健康人群血清中SARS-CoV抗体水平调查，以发现是否存在隐性感染，确定易感人群和高危人群，寻找与病源有关的人群和动物。方法 2003年8月～2004年1月，采集该地区各县区不同年龄、不同职业的健康人群血液，用酶联免疫法检测血清中SARS-CoV抗体Ig6水平。结果 共检测健康入血清4268份，SARS-CoV抗体Ig6阳性率为0．75％(32／4268)，其中1～6岁年龄组的阳性率为1．54％(15／975)，7～12岁年龄组的阳性率为1．23％(8／651)，13～18岁年龄组的阳性率为0．33％(1／306)，19岁～成年年龄组的阳性率为0．34％(8／2336)。2642份不同职业的健康人群阳性率为12．50％(3／24)，其他职业健康人群的阳性率为0．23％(6／2618)。结论 该地区SARS-CoV抗体水平比较低，低龄组(1～12岁)健康人群SARS-CoV抗体IgG阳性率高于高龄组(13岁～成年)组健康人群的阳性率；从事野生动物屠宰职业的健康人群SAILS-CoV抗体IgG阳性率高于其他健康人群的阳性率。提示SARS-CoV可能存在隐性感染、隐性传播和隐生感染聚集性；也提示从事野生动物屠宰职业的人群可能是SARS-CoV的高危人群。     

SARS-CoV infection in a restaurant from palm civet

Epidemiologic investigations showed that 2 of 4 patients with severe acute respiratory syndrome (SARS) identified in the winter of 2003-2004 were a waitresss at a restaurant in Guangzhou, China, that served palm civets as food and a customer who ate in the restaurant ashort distance from animal cages. All 6 palm civets at the restaurant were positive for SARS-associated coronavirus (SARS-CoV). Partial spike (S) gene sequences of SARS-CoV from the 2 patients were identical to 4 of 5 Sgene viral sequences from palm civets. Phylogenetic analysis showed that SARS-CoV from palm civets in the restaurant was most closely related to animal isolates. SARS cases at the restaurant were the result of recent interspecies transfer from the putative palm civet reservoir, and not the result of continued circulation of SARS-CoV in the human population.     

广州市某野生动物市场从业人员携带SARS冠状病毒抗体状况调查

目的了解广州市新源野生动物市场从业人员严重急性呼吸综合征（SANS）抗体产生情况及其暴露果子狸的关系。方法对野生动物市场从业人员中的志愿者进行SARS病毒血清抗体监测，对监测结果结合果子狸在市场的存在情况进行分析。结果2003年5和12月、2004年1和7月、2005年6月，共5次分别对328、238、135、139和53名市场从业人员开展调查，SARS病毒IgG抗体检出率依次为25．61％、13．03％、12．59％、5．04％和9．43％，没有检测到SARS病毒IgM抗体。在接受2次以上随访调查的129人中，97人的血清抗体一直为阴性，博人一直为阳性；有13人从阳性转为阴性，有1人从阴性转为阳性。在清除果子狸后进入市场的工作人员，SARS病毒IgG抗体检测全部阴性。结论广州市新源野生动物市场从业人员SARS病毒抗体检出率和市场内果子狸的存在情况密切相关。     

Prevalence of IgG antibody to SARS-associated coronavirus in animal traders--Guangdong Province, China, 2003

Severe acute respiratory syndrome (SARS) was identified in 2003 as an infectious disease caused by the SARS-associated coronavirus (SARS-CoV), a member of the coronavirus family not observed previously in humans. Because its sequence data differ from that of known human coronaviruses, SARS-CoV is suspected to have crossed the species barrier between an animal host and humans. The SARS outbreak began in China's Guangdong Province, where approximately 1,500 probable cases were identified during November 2002-June 2003. Detection of SARS-like coronavirus has been reported previously in masked palm civets (sometimes called civet cats) and a raccoon dog for sale in a live animal market in Shenzhen municipality. This report summarizes results of an investigation conducted by public health authorities in Guangdong Province, which compared the seroprevalence of SARS-CoV IgG antibody in animal traders (i.e., workers in live animal markets) with that of persons in control groups. The results indicated that 13% of the animal traders, none of whom had SARS diagnosed, had IgG antibody to SARS-CoV, compared with 1%-3% of persons in three control groups. Although the results provide indirect support for the hypothesis of an animal origin for SARS, they also underscore the need for detailed patient histories and more focused animal studies to confirm an animal origin for SARS.     

Antibodies to SARS coronavirus in civets

Using three different assays, we examined 103 serum samples collected from different civet farms and a market in China in June 2003 and January 2004. While civets on farms were largely free from SARS-CoV infection, approximately 80% of the animals from one animal market in Guangzhou contained significant levels of antibody to SARS-CoV, which suggests no widespread infection among civets resident on farms, and the infection of civets in the market might be associated with trading activities under the conditions of overcrowding and mixing of various animal species.     

Receptor-binding domain of SARS-CoV spike protein induces long-term protective immunity in an animal model

Development of effective vaccines against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) is still a priority in prevention of re-emergence of SARS. Our previous studies have shown that the receptor-binding domain (RBD) of SARS-CoV spike (S) protein elicits highly potent neutralizing antibody responses in the immunized animals. But it is unknown whether RBD can also induce protective immunity in an animal model, a key aspect for vaccine development. In this study, BALB/c mice were vaccinated intramuscularly (i.m.) with 10microg of RBD-Fc (RBD fused with human IgG1 Fc) and boosted twice at 3-week intervals and one more time at 12th month. Humoral immune responses of vaccinated mice were investigated for up to 12 months at a 1-month interval and the neutralizing titers of produced antibodies were reported at months 0, 3, 6 and 12 post-vaccination. Mice were challenged with the homologous strain of SARS-CoV 5 days after the last boost, and sacrificed 5 days after the challenge. Mouse lung tissues were collected for detection of viral load, virus replication and histopathological effects. Our results showed that RBD-Fc vaccination induced high titer of S-specific antibodies with long-term and potent SARS-CoV neutralizing activity. Four of five vaccinated mice were protected from subsequent SARS-CoV challenge because no significant virus replication, and no obvious histopathological changes were found in the lung tissues of the vaccinated mice challenged with SARS-CoV. Only one vaccinated mouse had mild alveolar damage in the lung tissues. In contrast, high copies of SARS-CoV RNA and virus replication were detected, and pathological changes were observed in the lung tissues of the control mice. In conclusion, our findings suggest that RBD, which can induce protective antibodies to SARS-CoV, may be further developed as a safe and effective SARS subunit vaccine.     

Immunogenicity and protective efficacy in monkeys of purified inactivated Vero-cell SARS vaccine

BACKGROUND: In 2003, severe acute respiratory syndrome (SARS) resulted in hundreds of infections and deaths globally. We aim to assess immunogenicity and protective efficacy of purified inactivated Vero-cell SARS vaccine in monkeys. METHODS: The cultures of SARS coronavirus (SARS-CoV) BJ-01 strain infected Vero cells were inactivated with beta-propiolactone. Sequential procedures, including ultrafiltration, gel filtration and ion exchange chromatography, were performed to obtain purified inactivated SARS vaccine. The purified SARS vaccine was analyzed with electron microscope, HPLC and Western blotting. We immunized three groups of cynomolgus macaques fascicularis with adjuvant-containing purified vaccine, purified vaccine and unpurified vaccine, respectively, and a fourth group served as a control. Antibody titers were measured by plaque reduction neutralization test. The vaccinated monkeys were challenged with SARS-CoV BJ-01 strain to observe protective efficacy. Additionally, three groups of rhesus monkeys were immunized with different doses of the purified inactivated SARS vaccine (0.5, 1 and 2mug/time/monkey) on days 0 and 7, and the monkeys were challenged with SARS-CoV GZ-01 strain. We assessed the safety of the SARS vaccine and observed whether the antibody dependent enhancement (ADE) occurred under low levels of neutralizing antibody in rhesus. FINDINGS: The purity of SARS vaccine was 97.6% by HPLC identification and reacted with convalescent sera of SARS patients. The purified SARS vaccine induced high levels of neutralizing antibodies and prevented the replication of SARS-CoV in monkeys. Under low levels of neutralizing antibody, no exacerbation of clinical symptoms was observed when the immunized monkeys were challenged with SARS-CoV. In this preliminary animal trial, no side effects were detected when monkeys were immunized with purified SARS vaccine either at normal or large doses. INTERPRETATION: The purified inactivated SARS vaccine could induce high levels of neutralizing antibody, and protect the monkeys from the challenge of SARS-CoV. The SARS vaccine prepared in the study appeared to be safe in monkeys.     

2017年泰州市重点职业病危害因素接触人员职业健康风险评估分析

依据《江苏省重点职业病监测与职业健康风险评估工作实施方案》要求，分析泰州市职业健康检查、职业病诊断与鉴定、职业病患者工伤保险待遇落实及职业病报告信息。结果显示，2017年职业健康检查共16759人，检出职业病10例，疑似职业病23例，职业禁忌证22人。职业病患者均已享受工伤保险待遇。泰州市重点职业病危害因素为电焊烟尘、矽尘和噪声，应进一步加强职业健康监护工作。