浙江省丽水市狂犬病新发疫区病毒检测及在防制中的应用

目的了解狂犬病毒在疫区犬间的流行状况。方法收集浙江省丽水市198份狂犬病新发疫区犬脑组织标本,用直接免疫荧光试验（DFA）特异性检测狂犬病毒抗原和逆转录-聚合酶链反应（RT-PCR）特异性检测狂犬病毒核酸,确定犬感染狂犬病毒状况。 结果犬脑组织DFA和RT-PCR平行检测阳性的标本13份,阳性率为6.57%（13/198）,其中松阳县9份,占采集标本的69.23%（9/13）,具有攻击人/犬史或者被犬攻击史的疑似狂犬病犬标本10份,占采集标本的76.92%（10/13）。 结论免疫学和分子生物学两种方法平行进行犬狂犬病毒的实验室检测,可以更准确地进行狂犬病的实验室诊断。市民暴露于犬后要及时到所在地狂犬病暴露门诊严格按照国家相关标准及规范正确处理伤口并注射狂犬病疫苗及抗血清或人源免疫球蛋白。建议相关部门加强犬的管理,同时对犬进行全面的狂犬病疫苗免疫接种。     

2006-2008年浙江省松阳县狂犬病流行因素调查

目的研究浙江省松阳县狂犬病的流行因素、流行规律。方法对狂犬病进行个案调查,同时用直接免疫荧光法和RT-PCR方法分别检测疫区234份宿主动物脑组织中的狂犬病毒抗原及核酸确定阳性标本。结果2006-2008年,松阳县共报告2例狂犬病。从234份宿主动物脑组织检测出狂犬病毒抗原阳性标本9份,阳性率为3.85%（9/234）,阳性者全部为犬;犬阳性率为8.26%（9/109）。结论犬只饲养量多,缺乏监管,且犬只免疫率极低,病毒感染率高,暴露后未进行伤口正确处理及免疫接种等是狂犬病发病回升的主要原因。有关部门应加强合作,在现阶段采取切实可行的预防控制策略和措施,遏止疫情的回升势头。     

湖南省疑似狂犬病病例皮肤组织标本首次检出狂犬病病毒

目的对疑似狂犬病病例标本进行狂犬病病毒抗原检测,为临床病例诊断提供实验室依据。方法对疑似病例组织标本（皮肤、角膜、脑组织等）运用直接免疫荧光实验法检测狂犬病病毒抗原。结果在送检的皮肤、角膜、脑组织中,皮肤、脑组织狂犬病病毒抗原阳性,角膜阴性。结论在疑似狂犬病病例的背部皮肤毛囊组织中首次检出狂犬病病毒,填补了湖南省疑似狂犬病病例皮肤标本实验室诊断的空白,为狂犬病疑似病例的早期实验室诊断提供了经验和依据。     

浙江省丽水市人感染H7N9禽流感病毒监测结果分析

目的了解H7N9禽流感病毒在人、禽间的分布情况,指导人感染H7N9禽流感防制工作。方法对2013年4月至2014年2月浙江省丽水市医疗机构发热伴呼吸道症状病例及外环境标本采用实时荧光定量聚合酶链反应方法检测H7N9亚型禽流感病毒核酸,对监测结果进行分析。结果轻症流感样病例和严重呼吸道感染病例分别检测768份和605份,H7N9禽流感病毒全部阴性,不明原因肺炎病例检测92份,阳性1份,阳性率1.09%;家禽粪便标本、笼具表面涂抹物、鸡蛋外壳涂抹物及不明原因死亡家野禽咽拭子和肛拭子共检测1317份,H7N9禽流感病毒阳性22份,阳性率1.67%,阳性标本分布在4个县(区)的8个活禽交易市场。结论首次在丽水市检测到人感染H7N9禽流感病毒,并在多个县(区)的活禽交易市场外环境检测到H7N9禽流感病毒,提示H7N9禽流感病毒已在丽水市禽间传播,存在人感染H7N9禽流感病毒的风险。     

湖南省狂犬病病毒种群划分情况及防控形势分析

目的 更新湖南省流行的狂犬病病毒（RABV）种群类型及进化特征,分析研判湖南省狂犬病疫情防控形势。方法 对湖南省2012—2017年收集的犬脑组织标本采用直接免疫荧光法或者反转录聚合酶链式反应法进行检测,对阳性标本进行N基因全序列（1 353 bp）测定及序列同源性分析,并与我国7个RABV种群的代表株一起进行系统发育分析。结果 新测的25株湖南省RABV毒株的N基因核苷酸同源性为91.9%～100%,氨基酸同源性为83.6%～100%;种群划分结果显示其中84%(21/25)的毒株为ChinaⅠ群,4株为ChinaⅡ群,未监测到2010年前在湖南流行的China Ⅲ群和China Ⅴ群。结论 湖南省RABV流行种群类型未增加,且局限于我国两个主要的犬源种群。切实加强犬的管理,尤其是农村犬的管理和免疫工作,才能进一步控制湖南省狂犬病疫情。     

上海市儿童下呼吸道感染常见病毒诊断方法比较

目的应用直接免疫荧光法（DFA）和多重逆转录聚合酶链反应（RT-PCR）对呼吸道感染患儿鼻咽分泌物的常见病毒进行检测,比较2种方法的符合率并评估其临床应用价值。方法选取急性呼吸道感染患儿鼻咽分泌物540例,分别用DFA和多重RT-PCR进行检测,分析其结果。结果 DFA检测阳性率为43.9%（237/540）,多重RT-PCR检测阳性率为55.7%（301/540）,多重RT-PCR阳性检出率显著高于DFA的阳性检出率（χ2=29.093,P〈0.01）。DFA和多重RT-PCR同时阳性163例,DFA阳性而多重RT-PCR阴性74例,多重RT-PCR阳性而DFA阴性138例,DFA和多重RT-PCR同时阴性165例。DFA和多重RT-PCR同时为阳性的163例标本中,有15例标本两者检测结果不一致,符合率为91.0%。结论 DFA快速、简便、特异性及敏感性高,适用于临床检测。多重RT-PCR的敏感性高,检测病毒谱广,且可以做亚型鉴定,适用于呼吸道病毒流行病学的调查。     

2010-2015年山东省狂犬病病毒分子流行病学特征

目的了解山东省境内流行的狂犬病病毒(RABV)的遗传进化特征和流行特点。方法对山东省2010-2015年收集的犬脑组织标本和病例唾液、血液等标本用RABV直接免疫荧光法或反转录-聚合酶链式反应法进行检测,检测阳性标本进行N基因序列测定和种系发生分析。结果 70份犬脑组织标本有11份为阳性,31份病例液体标本有2份为阳性,13份阳性标本都测序得到N基因全序(1 353 bp)。汇总已发表所有山东标本N基因序列(共26个)以及我国7个RABV种群的代表株共同构建的种系发生树显示,近90%的山东株(2006-2015年)属于ChinaⅠ进化群,其余3株(2006-2008年)属于ChinaⅡ进化群;山东省东部和西部地区的流行株相对聚集又明显交叉,泰安流行株最具多样性。结论山东省RABV分属ChinaⅠ和Ⅱ两个种群,近年山东省狂犬病的流行基本是ChinaⅠ传播扩散的结果。     

一起家庭聚集性发热伴血小板减少综合征疫情的流行病学及病原学分析

  目的   对一起家庭聚集性发热伴血小板减少综合征（SFTS）疫情的流行病学和病原学进行分析,为疾病防控及监测方案的制定提供参考。   方法   对2例SFTS病例开展流行病学调查,对其居家周围的人群及宿主动物进行监测,通过酶联免疫吸附试验（ELISA）和实时荧光定量PCR方法对病例、动物血清及蜱进行血清学和病原学检测,对病例和蜱进行疾病关联分析,对核酸阳性标本进行SFTS病毒的S、M、L基因测序,构建系统进化树。   结果   2例病例SFTS病毒IgM及总抗体均为阳性,其中1例核酸检测阴性。 45份健康人群血清标本中,3份SFTS病毒抗体阳性,6只犬（含病例家养犬）和4只羊血清SFTS病毒总抗体均为阳性,4头猪血清总抗体阴性。 10只蜱中,仅在病例家养犬寄生的蜱标本中检测到SFTS病毒核酸,基因序列测定显示SFTS病例株与蜱株均包含S、M、L片段,两者高度同源,核苷酸同源性为99.9%～100.0%。   结论   本起家庭聚集性SFTS疫情的发生与病例家养犬及其寄生蜱高度关联。     

多重RT-PCR与毛细电泳联用技术在儿童下呼吸道感染病原体检测中的应用

目的采用多重逆转录聚合酶链反应(RT-PCR)与毛细电泳联用技术对13种儿童呼吸道常见病原体痰液标本进行检测,研讨其在儿童下呼吸道感染病原体检测中的价值。方法采集2019年12月至2020年1月该院38例因下呼吸道感染住院患儿的痰液标本,应用多重RT-PCR技术检测13种呼吸道病原体,同时采用直接免疫荧光法检测7种常见呼吸道病毒抗原。结果38例下呼吸道感染住院患儿的痰液标本中,多重RT-PCR检出至少一种病原体阳性36份,检出率94.7%(36/38),其中乙型流感病毒检出最多,阳性20例(52.6%),其次检出呼吸道合胞病毒14例(36.8%)阳性,甲型流感病毒H1N1(2009)8例(21.0%)阳性,鼻病毒6例(15.8%)阳性,副流感病毒4例(10.5%)阳性,其中检出2种或2种以上呼吸道病原体混合感染14份,检出率为36.8%(14/38)。选取与直接免疫荧光法(DFA)检测相同的7种病毒进行比较,DFA检出阳性16份,检出率仅有42.1%(16/38),多重RT-PCR阳性检出率高于DFA方法,两种方法病毒阳性检出率差异有统计学意义(P<0.01)。结论多重RT-PCR与毛细电泳联用方法是一种高效快速、高通量、准确的检测技术;同DFA比较特别在流感病毒检测方面具有更高灵敏度,可同时检测更多病原体,能指导临床了解儿童下呼吸道感染病原并进行及时有效治疗。     

2012－2015年云南省狂犬病病毒糖蛋白基因进化特征分析

目的 了解云南省2012－2015年35株狂犬病病毒（RABV）糖蛋白（G）基因的进化特征及流行病学特点。方法 在云南省狂犬病流行区采集可疑犬脑组织以及狂犬病患者脑组织和唾液标本,用直接免疫荧光法进行病毒抗原检测,阳性标本提取病毒核酸,通过反转录聚合酶链式反应（RT-PCR）扩增RABV的G基因序列,用MEGA 5.0软件邻接法（NJ）进行系统进化树分析。结果 经RT-PCR和序列测定获得2012－2015年云南省35株RABV的G基因序列,并与2006－2011年获得的云南省11株RABV及邻省和东南亚地区的15株RABV的G基因序列进行系统进化分析。结果表明,云南省46株RABV中,除2006年的Tc06株属于China-Ⅵ进化群外,其他45株均属于China-Ⅰ进化群。云南省China-Ⅰ进化群流行株在2006－2015年间每年均有分布,其中,2006－2011年10株,2012－2015年35株,分布于云南省10个州（市）并与来自四川、贵州和湖南省的China-Ⅰ进化群病毒株的进化关系最为接近。China-Ⅵ（Tc06）则与缅甸、老挝和越南的东南亚进化群病毒株亲缘关系最近。结论 2012－2015年,RABV China-Ⅰ进化群在云南省狂犬病流行区广泛分布,属云南省主要进化群并具较强的传播速率,期间云南省未再发现China-Ⅱ、China-Ⅲ和China-Ⅵ等进化群病毒株的流行。     

云南省禄丰县家犬狂犬病毒带病毒率及阳性犬分布情况的调查

目的 分析云南省禄丰县自然状态家犬狂犬病毒携带阳性率及阳性犬分布情况,为制定狂犬病防制对策提供依据。 方法 2012年3月12 18日在全面扑杀8个自然村疫区的犬只过程中,采犬脑标本检验,调查家犬狂犬病毒携带阳性率,分析阳性犬分布情况;全面扑杀犬只3个月、6个月和1年后开展3次农户恢复养犬情况调查。 结果 8个自然村扑杀1103只家犬,采集犬脑标本542只(份),检测阳性24只,阳性率4.43%,阳性犬呈离散性分布,阳性率最高的自然村达9.09%;全面扑杀犬只1年后养犬户和养犬数分别恢复到扑杀前的80.38%和61.92%,户均养犬恢复到0.56只(扑杀前0.90只)。 结论 禄丰县家犬狂犬病毒带病毒率不高,阳性犬呈离散性分布,阳性率与疫点距离远近不存在比例关系。扑杀后农户恢复养犬较快,犬只来源较为复杂,狂犬病等疫源输入的危险因素增加。 科学抽样监测动物狂犬病带病毒动态,以免误杀健康犬只,可避免大面积扑杀犬只带来的社会慌恐和经济损失。     

Retrospective study on the possibility of an SFTS outbreak associated with undiagnosed febrile illness in veterinary professionals and a family with sick dogs in 2003

IntroductionSevere fever with thrombocytopenia syndrome (SFTS) is an emerging tick-born disease and its animal-to-human transmission has come to attention recently. During our sero-survey of SFTS virus (SFTSV) among veterinary professionals in 2018, a veterinarian and his assistant working in an animal hospital were tested positive by enzyme-linked immunosorbent assay (ELISA). An additional survey implied a cluster of SFTS cases in which four more people, a family who brought two sick dogs to the animal hospital in 2003, were involved. This study aimed at assessing the possibility of animal-to-human transmission of SFTSV in this cluster.MethodsRetrospective interviews were performed with the owner family of the dogs and their clinical records were obtained from each hospital. SFTSV-IgG were tested by ELISA and virus neutralization test using the sera collected from them in 2018.ResultsThe interviews revealed that a total of six people, the two veterinary professionals and the owner family who took care of the sick dogs, suffered from SFTS-like symptoms in the same period of time in 2003. All patients did not have tick bite before the onset and all suspected causative agents were excluded by laboratory tests. The serological tests in this study revealed the four owner family members were all positive for SFTSV antibodies.ConclusionsConsidering the extremely low seroprevalence of SFTSV antibodies among inhabitants of the region, the existence of SFTSV antibodies in all these six people presents a possibility that they were involved in an SFTS outbreak originated in the sick dogs in 2003.     

Acute disseminated encephalomyelitis following immunization with homologous brain extracts; studies on the role of a circulating antibody in the production of the condition in dogs

1. A severe demyelinating condition characterized by ataxia and paralysis, in some instances leading to death, was produced in thirty-five of a total of fifty-five dogs following immunization with homologous brain tissue combined with Freund's adjuvants. In more than 30 per cent of instances paralysis did not occur until immunization was continued for 6 or more months. Only eight dogs became paralyzed after a single injection of antigen. The condition appeared between 6 and 15 days after the last injection in all animals, irrespective of the total number of injections or the duration of immunization. 2. An antibody which reacted in complement fixation tests with aqueous and alcoholic extracts of homologous brain tissue was demonstrable in the majority of immunized dogs, whether or not the animals became paralyzed. It appeared during or after the 3rd week of immunization, and its occurrence or titer could not be correlated with the incidence of the encephalomyelitis. In general, there were fewer dogs with demonstrable antibody in the paralyzed group than in the non-paralyzed group. 3. A flocculation reaction with alcohol extracts of homologous brain was demonstrated in the serum of immunized dogs. The antigen and antibody involved were apparently identical with those responsible for the complement fixation reactions. 4. The brain tissue component which reacted as antigen in the complement fixation test was present in adult brain from several mammalian species, and peripheral nerve. It was not present in the brain of newborn dogs nor in other unrelated organs. It was demonstrable in brain tissue which had been allowed to autolyze, or treated with 10 per cent formalin. It was not impaired by boiling, or by acid hydrolysis, and was contained in the unsaponifiable fraction of brain lipids. It was separable from cholesterol by digitonin precipitation of the latter. 5. Immunization of dogs with the unsaponifiable fraction of homologous brain, in adjuvants, caused the appearance of antibrain antibody similar to that in animals injected with whole brain. Encephalomyelitis was not observed during a 2 month period of immunization with this fraction. 6. In guinea pigs, an injection of the unsaponifiable fraction of brain, in adjuvants, was followed by fatal meningoencephalitis, but the identity of the state with that caused by whole brain antigens was not established.     

Disposition in the dog and the rat of 2, 6-diamino-9-(2-hydroxyethoxymethyl)purine (A134U), a potential prodrug of acyclovir

2,6-Diamino-9-(2-hydroxyethoxymethyl)purine (A134U), the 6-deoxy-6-amino analog of the antiviral agent acyclovir (ACV), was administered orally to dogs and rats. Plasma concentration-time profiles and urinary excretion of A134U and its deamination product, ACV, were determined. Mean peak plasma ACV concentrations achieved in the dog were 57, 156 and 285 microM after A134U doses of 20, 50 and 120 mg/kg, respectively, and increased in near proportion to the dose. The urinary recovery of ACV accounted for 60-92% of the two lower doses, but only 40-58% of the highest dose. In the rat, peak plasma ACV concentrations were 3.1 and 10.7 microM, respectively, after 20- and 50-mg/kg doses of A134U. After 5- and 20-mg/kg oral doses of [2-14C]A134U, the urinary recovery of ACV (20-27%) accounted for 59 to 76% of the absorbed dose. The remainder was excreted largely as unchanged A134U, with negligible (0.4-1.3%) biotransformation to inactive metabolites. Except for small decreases in absorption and increases in deamination, no change in the metabolism of A134U was observed after its repeated oral administration to rats. Oral dosing of dogs and rats with A134U resulted in peak plasma concentrations and total urinary recoveries of ACV greater than those observed after equivalent oral doses of ACV, suggesting that A134U might be an effective prodrug of ACV for use in the oral therapy of herpes simplex virus infections.     

2011年浙江省金华市狂犬病监测结果分析

目的 分析浙江省金华市2011年狂犬病监测资料,评价预防处置效果,为预防和控制狂犬病提供依据。 方法 根据《浙江省狂犬病监测方案》要求,收集全市狂犬病监测资料,采用描述流行病学方法进行分析。 结果 2011年金华市报告狂犬病4例,死亡4例,4例病例唾液标本经检测均为狂犬病毒核酸阳性。报告狂犬病暴露病例60 866例,暴露率为1474.61／10万。病例时间分布有明显的季节性,呈夏季高发特点。以青壮年为主,20~59岁病例占总数的57.88%;暴露率以0~9岁年龄段最高,为2322.58/10万。病例以农民最多,占总数的66.34%。伤人动物最多的为犬,占92.40%。Ⅲ级暴露者比例为18.18%,狂犬病免疫球蛋白接种率为21.67%,且各县(市、区)差异较大。报告一犬伤多人事件6起,采集5只犬脑组织经检测均为狂犬病毒核酸阳性。 结论 金华市狂犬病防控形势严峻,狂犬病防制工作仍有待加强。