延吉市成人腹泻爆发流行的病原学研究

1986年5月延边地区4个县市腹泻爆发流行,发病涉及各年龄组,但以青壮年人为主。取22名患者粪便经细菌培养,未发现致病菌,而有50%(11/22的粪便标本在电镜下观察到52~68nm左右的轮状病毒颗粒,85.7%(6/7)的粪便标本成人轮状病毒ELISA检测呈阳性,59%(13/22)的粪便标本病毒核酸聚丙烯酰胺凝胶电泳检测阳性,病毒核酸图象呈现轮状病毒特有的11条核酸带。因此,可以确定这次流行性腹泻爆发流行的病原是成人腹泻轮状病毒,发现成人轮状病毒,在吉林省是首次。     

青海省67份轮状病毒结构蛋白VP7基因分型和分布情况

目的 研究2016-2017年青海地区腹泻患者轮状病毒基因分型及流行病学分布。方法 收集门诊及住院部腹泻的粪便标本238份,采用实时荧光PCR法对轮状病毒A组进行检测,阳性标本进行VP7基因扩增和测序。结果 238份粪便标本中,通过实时荧光PCR 检测到轮状病毒A组阳性67份,阳性率为28.15%（67/238）;对67份轮状病毒A阳性标本进行VP7蛋白检测和测序,测序后得到29份核苷酸序列,用Clustral X Bootstap NJ Tree软件构件进化树,分析发现2016年3月-2017年12月青海轮状病毒以G9P8型为主,共26株,占89.66% (26/29),G2P4型2株(2/28),G3P8型1株(1/28), 轮状病毒腹泻发病高发季节为9-12月,其中以12月份检出最高,占总数的61.19%（41/67）。病人以成人为主,成人和5岁以下儿童比例为1.73[DK]∶1。结论 2016-2017年青海地区轮状病毒以流行病毒株G9P8型为主。     

一株新轮状病毒引起河北省石家庄市成人腹泻爆发流行

1997年 4月 10～ 2 8日 ,在河北省石家庄市某高校发生一起成人急性腹泻的爆发流行 ,累积10 0 0多名大学生发病。经不连续聚丙烯酰胺凝胶 (PAGE)电泳检查腹泻病人粪便标本核酸图谱一致呈4 - 2 - 1- 1- 1- 1- 1排列的新轮状病毒 (新RV) 14份 ,阳性率占 4 7% (共检标本 30份 ) ,且未见其它带型的轮状病毒。提示该核酸图谱的轮状病毒是此次成人急性腹泻爆发流行的主要病原。用已知ADRV第 5及第 9基因片段末端引物进行逆转录 -聚合酶链反应 (RT -PCR) ,结果阳性对照ADRV的扩增结果都为阳性 ,而新RV的扩增结果都为阴性。这些结果表明该新RV不属于B组轮状病毒 ,是与成人腹泻轮状病毒 (ADRV)完全不同的新RV。非ADRV的轮状病毒引起成人腹泻爆发流行在河北省尚属首次。     

腹泻患者粪便标本中A组轮状病毒感染特征分析

目的:分析病毒性腹泻患者中人轮状病毒(Human Rotavirus,HRV)的感染情况及毒株流行特点。方法:收集杭州市239份疑似病毒性腹泻患者粪便标本分别应用酶联免疫吸附法及RT-PCR法进行HRV抗原及核酸检测。。其中HRV阳性核酸应用巢式RT-PCR进行轮状病毒分型检测。结果:239份粪便标本中HRV抗原阳性为44份(18.4%),HRV核酸阳性为55份(23.0%),其中G血清分型:G3型15份(27.3%),G1型9份(16.4%),G2型为7份(12.7%),G9型为1份(1.8%),G2+G3混合感染1份(1.8%),未分型22份(40%)。结论:A组轮状病毒是婴幼儿病毒性腹泻的重要病原体,同时也可引起成人腹泻散发流行,成人A组轮状病毒流行季节和流行株与婴幼儿一致,G3为主要流行株,且同时存在多种血清型流行,未分型标本的比例较大有待深入研究。     

2011—2015年北京西城区成人门诊腹泻患者诺如病毒与轮状病毒感染的流行病学特征

目的了解门诊腹泻病例中诺如病毒和轮状病毒感染现状,分析本辖区病毒感染性腹泻的流行特征。方法以2011—2015年在北京市西城区两所三甲医院成人肠道门诊就诊的腹泻病例作为监测对象,对其进行诺如病毒和轮状病毒检测及流行病学调查分析。结果调查检测腹泻病例共714例,200例检测阳性,总阳性率28.01%;其中轮状病毒检测阳性率为10.08%;诺如病毒检测阳性率为18.63%。诺如病毒检测阳性率显著高于轮状病毒阳性率(χ~2=7.211,P<0.05)。诺如病毒检测阳性病例中有腹泻10次以上、恶心、呕吐、水样便症状的病例百分比明显高于检测阴性病例;轮状病毒检测阳性病例中有腹痛症状的病例百分比明显低于检测阴性病例,腹泻10次以上的病例百分比明显高于检测阴性病例;诺如病毒及轮状病毒检测阳性病例中,便常规检查分别有白细胞和红细胞的病例百分比均显著低于检测阴性病例,差异有统计学意义。结论北京市西城区成人肠道门诊就诊的腹泻患者中诺如病毒和轮状病毒感染较为普遍,腹泻多次水样便、恶心、呕吐是诺如病毒感染的常见症状;次数较多的无腹痛腹泻是轮状病毒感染的常见表现。     

苏州市婴幼儿轮状病毒腹泻分子流行病学研究

目的 研究苏州市5岁以下婴幼儿轮状病毒腹泻分子流行病学特征。方法 对2001年9月～2002年8月期间在苏州大学附属儿童医院就诊的5岁以下腹泻患儿进行调查。收集腹泻患儿粪便标本检测轮状病毒并对轮状病毒阳性标本用ELISA、PCR法进行分型研究。结果 共检测标本775份，轮状病毒阳性274份，检出率35．35％；血清分型发现，轮状病毒腹泻G分型以G3(40．15％)和G1(28．42％)为主要流行株，基因P分型结果以P[4](39．68％)为主，尚发现较多不常见G／P组合和未分型病毒株。结论 苏州市婴幼儿轮状病毒腹泻，以G3和P[4]最多见，但有较多未分型病毒株，应继续进行轮状病毒腹泻的监测。     

北京市成人诺如病毒性腹泻流行病学调查

目的探讨北京市成人诺如病毒性腹泻的流行病学特点。方法采集2007年10月~2008年1月在中日友好医院就诊的232例成人急性腹泻患者的粪便样本,用酶联免疫吸附试验(ELISA)检测粪便诺如病毒抗原。统计学处理采用SPSS 10.0统计软件,率的差异用χ2检验,P0.05为差异有统计学意义。结果232份粪便样本中诺如病毒抗原阳性136例,阳性率为58.62%,各年龄组间阳性率差异无统计学意义(P0.05)。10月份诺如病毒检出率最高,为63.30%(69/109),12月份检出率最低,为44.44%(20/45),差异有统计学意义(P0.05)。136例诺如病毒阳性患者中,出现呕吐症状者占47.79%(65/136),而诺如病毒阴性患者为26.04%(25/96),差异有统计学意义(P0.001)。结论本调查研究表明,诺如病毒是北京市秋冬季成人散发性腹泻的主要病原体,成人诺如病毒性腹泻患者易出现呕吐症状。     

2012-2013年上海市浦东新区成人病毒性腹泻感染情况调查

目的了解上海市浦东新区2012—2013年间成人病毒性腹泻感染情况。方法通过实时荧光PCR方法检测该区14家监测医院成人腹泻患者粪便标本中的轮状病毒、诺如病毒、星状病毒、扎如病毒和肠腺病毒。结果共检测腹泻患者粪便样本3 403份。在1 060份样本中检出至少1种病毒核酸,核酸阳性检出率为31.15%。诺如病毒、轮状病毒、扎如病毒、星状病毒以及肠腺病毒的检出率分别为21.28%、7.70%、1.85%、1.55%和0.59%。患者的性别分布不存在差异,性别比为0.95∶1。诺如病毒在各年龄组均有检出。不同病毒病原的感染存在不同的高峰期,但主要集中在秋冬季。结论诺如病毒是浦东地区成人病毒性腹泻的主要病原。应该加强对病毒性腹泻的监测,预防由诺如病毒引起的腹泻暴发。     

2009年长春地区婴幼儿病毒性腹泻的病原学监测分析

目的研究2009年长春婴幼儿病毒性腹泻的病原学特点。方法 2009年1-12月在长春市儿童医院共采集腹泻患儿的粪便标本419份,对其进行轮状病毒、杯状病毒、肠道腺病毒和星状病毒的检测分析。结果 419份标本中,采用ELISA法检测出轮状病毒184例,阳性率为43.91%(其中G基因型分型151例,以G3型为主,占64.24%);P基因型分型118例(其中以P8型为主,占92.37%);G/P优势组合型以G3P8为主,占48.75%(78/160)。采用RT-PCR法检测杯状病毒阳性67份,阳性率为15.99%;肠道腺病毒阳性8份,阳性率为1.91%;星状病毒阳性8份,阳性率为1.91%。结论轮状病毒是2009年长春地区婴幼儿病毒性腹泻的主要病原体,主要血清型为G3P8型;杯状病毒、星状病毒和肠道腺病毒也是重要的病原体。     

轮状病毒腹泻小儿特异性体液免疫应答的研究

目的 通过对轮状病毒腹泻小儿特异性体液免疫应答的研究,为有效预防和治疗轮状病毒感染提供理论依据.方法 75例粪轮状病毒检测阳性患儿(腹泻组)及45例近期无腹泻和其他感染性疾病小儿(对照组),采用ELISA法检测抗原及血浆和粪标本中的特异性抗体;采用反转录PCR法确定轮状病毒基因型;采用荧光定量PCR法进行外周血单个核细胞细胞因子mRNA表达的检测.结果 腹泻组轮状病毒G分型以G3型(77.3%,58/75)为主,P分型以P8型(82.7%,62/75)为主;腹泻组血浆中特异性抗体滴度均明显高于对照组,粪标本中IgA抗体滴度的增高尤为显著;腹泻组患儿CD19+细胞比率为(30.8±7.9)%,显著高于对照组的(23.1±7.7)%(P=0.009).CD4+细胞比率明显下降(P=0.005),IL-12p40 mRNA的表达水平在疾病全程均比对照组明显升高(P＜0.01).结论 轮状病毒感染小儿免疫应答以特异性体液免疫,尤其是黏膜免疫的显著增强为主要特点.     

Evaluation of a new assay kit for intrinsic factor blocking antibody (type I) as an aid in the diagnosis of pernicious anemia

Sera from 39 patients with pernicious anemia and 251 patients with various other diseases and 42 healthy normal subjects were tested for intrinsic factor blocking antibody by a Corning commercial kit. Twenty-five (64%) out of 39 pernicious anemia patients showed a positive reaction to the intrinsic factor blocking antibody. The positive incidence of intrinsic factor blocking antibody by this method agreed well with the results obtained in this laboratory over the past 10 years using the charcoal method. No sera from normal subjects tested were positive for the intrinsic factor blocking antibody. All 226 sera with vitamin B12 levels less than 3,500 pg/ml which came from patients without pernicious anemia, were negative for the intrinsic factor blocking antibody. In 15 sera with B12 levels greater than 3,500 pg/ml from patients who received recent B12 medications, 11 sera showed false positive results. On the other hand, no false positive results were obtained by this method in 10 sera with endogenous serum B12 levels greater than 3,500 pg/ml collected from patients with chronic myelogenous leukemia. It is reasonable to presume that this assay is clinically useful for detection of intrinsic factor blocking antibody in the diagnosis of pernicious anemia, if false positive results due to B12 medication are excluded.     

Antibody to serotype 8 rotavirus in Ecuadorian and German children.

Only 2 out of 71 German patients infected with rotavirus (3%) and 8 out of 147 German control patients (5%) showed serum antibody to the new serotype 8 rotavirus. Such antibody was detected in the sera of 232 of 870 Ecuadorian children (27%). Twelve Ecuadorian sera showed neutralizing activity only against serotype 8 and not to the other serotypes (1-4) tested, indicating that human serotype 8 rotavirus circulates in South America.     

An epidemic of rotavirus diarrhoea in Jawhar Taluk, Thane district, Maharashtra, India, December 2000-January 2001

An epidemic of diarrhoea in Jawhar, a tribal area of Thane district, Maharashtra, India was investigated. Within a period of approximately 2 months 490 cases of acute diarrhoea were reported among children under 5 years of age, with a case fatality rate of 0.40%. Twenty-seven out of 39 (69.23%) rectal swabs/faecal specimens obtained from hospitalized paediatric patients up to 2 years of age from Jawhar were positive by ELISA for rotavirus. Of these, seven were in the age group of < or = 6 months. Seven ELISA-positive faecal specimens were positive for serotype G3 by RT PCR. Out of 15 serum samples collected from these patients, 12 showed the presence of rotavirus-specific IgM. Rotavirus appears to be the aetiological agent of this widespread outbreak in Jawhar, Thane district, Maharashtra state, India.     

Nasal immunisation with Salmonella typhimurium producing rotavirus VP2 and VP6 antigens stimulates specific antibody response in serum and milk but fails to protect offspring.

Rotavirus specifically infects the small intestine of young infants resulting in severe diarrhoea. Mucosal antibody responses are required to cure the infection, and mucosal administration of rotavirus-like particles induces protective immunity without requiring a mucosal adjuvant such as cholera toxin. In addition, the rotavirus protein VP6 has been defined as a protective antigen in an adult mouse rotavirus infection model. Salmonella typhimurium is an epithelium-invasive bacterium that induces specific immune responses in mucosal tissues against itself and carried antigens. In this work, we investigated the capacity of a live recombinant S. typhimurium vaccine to stimulate antibody responses against rotavirus. We constructed an attenuated S. typhimurium strain simultaneously producing VP6 and VP2 rotavirus proteins in the cytoplasm. In contrast to expression in eukaryotic cells, VP6 and VP2 did not form virus-like particles in our bacterial system. After nasal administration of female mice, the live recombinant Salmonella were able to elicit an antibody response specific to both VP2 and VP6 in serum and milk. However, these antibodies failed to passively protect the offspring against rotavirus-induced diarrhoea.     

轮状病毒感染的血清流行病学调查

本文用对流免疫电泳法,检查各年龄组血清193份,阳性率67.4%,脐血14份,阳性率71.4%,首次报告轮状病毒感染在南通市人群中有相当的普遍性。并说明用此法进行轮状病毒感染的血清流行病学调查是一种操作简单,结果较为满意,且易于推广的方法.     

风疹病毒E1蛋白与DsbA蛋白原核融合表达及应用

目的 利用DsbA蛋白分子伴侣性质,原核系统内可溶性表达风疹病毒E1蛋白并进行纯化,用于检测风疹病毒特异性抗体IgM. 方法 将PCR扩增的风疹病毒E1核酸序列克隆至融合表达载体pET-DsbA中,依次进行表达和亲和纯化,利用产物建立IgM捕获ELISA方法鉴定DsbA-E1融合蛋白的抗原性及实用性. 结果 表达纯化的E1蛋白经酶标记建立IgM捕获ELISA方法,检测60份抗风疹病毒IgM阳性血清和60份健康人血清,其中阳性血清检出例数为45例,检出率为75％;健康人血清检出1例,检出率为1.7％,特异度为98％;用酶标记E1蛋白建立的捕获ELISA法阳性检出率98.3％ (59/60),阴性检出率100％,初步表明E1蛋白抗原表位有较好的抗原特异性. 结论 融合蛋白DsbA-E1在大肠杆菌中以可溶性表达形式存在,该重组蛋白抗原性强,利用其建立的IgM捕获ELISA方法,可用于检测风疹病毒抗体.     

Validation of an indirect immunoenzyme assay for the detection of antibodies against Trypanosoma evansi in horses in Argentina

An indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies against Trypanosoma evansi was evaluated using 90 different sera, obtained from naturally-infected horses. As negative controls, 218 sera from the T. evansi-free zone of Argentina, and 90 uninfected sera from the enzootic zone were used. The results of the ELISA were expressed in terms of percent positivity (PP) when compared with a positive primary reference serum, obtained from a horse experimentally-infected with T. evansi. The inter-assay coefficient of variation (CV), expressed as PP, was 44.7% for the negative control serum, 8.8% for the mildly positive reference serum, and 9.2% for the secondary positive control serum, while the intra-assay CV for each of the above sera was 6%, 2.8% and 5%, respectively. Positive and negative serological results were differentiated using a histogram of the distribution of the results obtained using sera from infected and uninfected animals from the enzootic zone (expressed in PP). A PP of 50 indicated a sensitivity of 95.5% for a confidence interval (CI) of 91.3% to 99.7%, and a specificity of 98% for a CI between 95% and 100%. Positive and negative predictive values were established for each rate of prevalence between 0.01% and 25%. The use of reference control sera in each assay enabled reproducible results to be obtained. The author recommends that this methodology be used whenever certification of the T. evansi status of horses is required, and particularly when animals are to be moved from an infected to a disease-free area.     

Studies of smallpox antibody levels of sera from samples of the vaccinated adult population of Madras

In view of the endemicity of smallpox in Madras and the poor take rate on revaccination with the lymph in current use, it was felt that the immunity of the general adult population might be relatively low. In an attempt to obtain some measure of this, sera were obtained from over 300 adults in Madras for the estimation of serum antibody. The results of neutralization tests against variola virus made on these sera showed that about 10% of those who had good scars from infant vaccination showed little or no antibody. A comparison of the antibody titres of adult sera showing good antibody levels when used undiluted was made with that of sera from smallpox convalescents. The antibody level in the latter sera was 20-100 times that of the sera of adult persons who showed good vaccination scars.     

Heterotypic protection from rotavirus infection in mice vaccinated with virus-like particles

Virus-like particles (VLPs) composed of rotavirus VP2, VP6, and VP7 of G1 or G3 serotype specificity were produced in insect cells coinfected with recombinant baculoviruses expressing single rotavirus genes. The VLPs were purified and subsequently evaluated for immunogenicity and protection in the adult mouse model of rotavirus infection. Mice were vaccinated twice intramuscularly with G1 VLPs formulated with Quillaja saponaria (QS-21) or adsorbed to aluminium hydroxide (AlOH), or with G1 VLPs alone. G3 VLPs, G1 plus G3 VLPs, inactivated SA11 virions formulated with QS-21, or adjuvants were similarly inoculated as controls. Mice were examined for serum and fecal antibody responses by ELISA or microneutralization assays. Protective efficacy of the VLP vaccine formulations against oral challenge with the G3 murine ECwt rotavirus was assessed by comparing the antigen shed in stool of the VLP-vaccinated mice to that of the adjuvant-immunized mice. G1 VLPs in QS-21 induced significantly higher serum and intestinal antibody titers than G1 VLPs in AlOH or G1 VLPs alone. QS-21 also heightened serum and fecal antibody responses to G3 VLPs. These QS-21-augmented antibody responses were further characterized by equivalent IgG1 and IgG2a titers in sera, suggesting that G1 or G3 VLPs in QS-21 induced a balanced Th1/Th2 response. G1 VLPs in QS-21 induced partial protection (88%) against oral challenge with the heterotypic ECwt virus, whereas G3 VLPs in QS-21 induced complete protection (100%). In contrast, G1 VLPs when formulated with AlOH induced a predominant Th2 response and did not protect (1%) mice from virus challenge. Our results indicate that the type of adjuvant used clearly influences both antibody responses to rotavirus VLPs and the protective efficacy against rotavirus infections. These data have important implications for the development of parenteral vaccines to ameliorate rotavirus disease.     

巴楚县2001年新疆出血热疫情的血清学证实

目的：以血清流行病学方法调查新闻出血热（XHF）病人，易感人群和主要宿主动物中疾病的感染情况。方法：分别收集2001年4－6月新疆巴县临床诊断为XHF的病人血清，易感人群血清和主要宿主动物的血清，用研制的诊断试剂以酶联免疫吸附试验（ELISA）检测XHF特异性IgG和IgM抗体；用抗原捕获ELISA检测XHF病毒抗原。结果：病人血清IgG抗体阳性率为39．62％（21／53）。IgM抗体阳性率为20．75％（11／53），抗原获ELISA有1份血清为XHF抗原阳性；易感人群血清IgG抗体阳性主为21．05％（4／19），IgM抗体检测和抗原捕获ELISA全部为阴性；羊血清IgG抗体阳性率为70％（56／80）。结论：血清流行病学研究证实该次疫情确系XHF、流行地区人畜均有较高水平的隐性感染。