快速检测成人腹泻轮状病毒(ADRV)抗原和抗体的研究

本文报道应用反相间接血凝和间接血凝试验技术快速检测ADRV抗原和抗体,一般在60分钟左右作出诊断。用ELISA方法和协同凝集试验作对照,结果一致。本实验与其它肠道病毒不发生交叉凝集反应,与菌痢、肠炎病人粪便和正常人粪便标本也不发生非特异性反应。实验准确可靠。本实验用间接血凝试验技术快速检测了ADRV抗体,观察了流行性腹泻病人血清中ADRV抗体持续时间。发现流行性腹泻患者发病后ADRV抗体存在半年以上,八个月有不同程度的下降,流行地区的健康人的ADRV抗体水平占33％左右,非流行地区正常人的ADRV抗体占11.3％左右。并用血凝抑制试验检测ADRV抗体,与小儿轮状病毒抗体无交叉反应。本试验对流行性腹泻的流行病学研究和临床诊断具有实用意义。     

成人腹泻轮状病毒ELISA方法的建立和应用

本文通过特异性试验、阻断试验、交叉试验、敏感度试验和重复性试验,建立了成人腹泻轮状病毒一酶联免疫吸附试验法(ADRV—ELISA)。应用此法检测了全国20多个省区202份病人腹泻标本,检出率为91％。采用本ELISA、核酸电泳、电镜三种方法对48份病人腹泻标本进行了双盲法检测比较,结果三种方法的阳性检出率分别为100％、85.4％、56.25％(P<0.05)。实验结果表明,本ELISA应用于检测成人腹泻轮状病毒(ADRV),具有敏感度高。特异性强等优点。     

湖北成人轮状病毒性腹泻爆发流行的病原学研究

1986年5月4日—5月19日,湖北省襄阳县发生七起以成人为主的流行性腹泻,发病98例,患病率为47.1％。发病涉及各年龄组,以青壮年为主。取18名患者粪便作细菌培养,未发现致病菌。而有50％(9/18)的粪便际本在电镜下观察到约75nm的轮状病毒颗粒;72.2％(13/18)的粪便标本聚丙烯酰胺电泳(PAGE)检测呈阳性,病毒核酸图象与普遍轮状病毒核酸图象十分不同;患者双份血清补体结合试验表明57.1％的病人恢复期血清产生对分离株轮状病毒的抗体。因此,可以确定,本次流行性愎泻爆发流行的病原是成人轮状病毒(ADRV)。     

成人腹泻轮状病毒的分子流行病学调查

本文在我们先前研究的基础上,报道用聚丙烯酰胺凝胶电泳技术,对我国12个地区53份成人腹泻轮状病毒(Adull Diarrhea Rotavirus, 简称 ADRV)的核酸(RNA)进行了分析。研究结果表明,我国各地流行的ADRV具有共同而独特的RNA图型,同时首次观察到不同地区的成人蝮泻轮状病毒RNA在电冰迁移率上存在着微小的差别;这意味着我国目前流行的AI)R、为一群基因图型基本相似,而在毒株间存在着微小的变异。     

在我国患急性腹泻成人中发现一种新轮状病毒

从3例急性成人腹泻患者粪便中,经电镜观察,成人腹泻轮状病毒—酶联免疫吸附试验(ADRV-ELISA)、普通轮状病毒—酶联免疫吸附试验(Rota-ELISA),猪抗C型(组)轮状病毒和鸡抗D型(组)轮状病毒血清分别与本病毒的免疫电镜试验以及RNA电泳等实验结果,发现了一种新轮状病毒,该病毒的形态结构与普通轮状病毒(Rotavirus)、成人腹泻论状病毒(Adult Diarrhoea Rotavirus,简称ADRV)极为相似,但抗原性与普通轮状病毒(A组),成人腹泻轮状病毒(B组),C型轮状病毒(C组),D型轮状病毒(D组)显然无关。基因分析表明,该病毒基因组由11个双链RNA片段组成,但其RNA电泳图型具有独自的特点,与目前公认的A组、B组、C组、D组论状病毒韵RNA电泳图型均不同,免疫电镜证实,该病毒能被人恢复期血清所凝集,表明该病毒可能是腹泻病人的病因。     

流行性成人腹泻病人排出成人腹泻轮状病毒的持续时间

成人腹泻轮状病毒(Adult Diarrhoea Rotavirus,简称ADRV)是引起青壮年急性胃肠炎的主要病原之一,1983年至1985年在我国许多地区暴发流行,严重危害着人民健康,至今尚无治疗药物。因此,摸清病人排病毒情况或规律,对及时控制传染源,预防ADRV大量传播有着重要意义。现将本文实验结果报告如下。     

成人腹泻轮状病毒的提纯及其高价免疫血清的制备

本文采用交叉免疫电泳技术,首先提纯了成人腹泻轮状病毒(Adult DiarrhoeaRotavirus简称:ADRV),并经电镜确认,用作免疫抗原,首次制备了高价兔与豚鼠抗ADRV血清和小鼠抗ADRV腹水,其特异性经交叉免疫电泳和对流免疫电泳鉴定,其效价经酶联免疫吸附试验(ELISA)检测均在1:2,000以上。     

仔猪副轮状病毒(C群)的检出

利用聚丙烯酰胺凝胶电泳法,从某猪场的20份仔猪腹泻标本中检出具有副轮状病毒(C群)RNA电泳图型者11份,其检出率高达55％。该病毒经口服接种未吮初乳的新生仔猪,可致急性水样腹泻,并排出大量病毒。经轮状病毒(A群)ELISA和副轮状病毒(B群)对流免疫电泳检测证明,它与轮状病毒(A群)和副轮状病毒(B群)均无血清学交叉。本文首次证实在我国猪群中存在有与国外报道的相同的猪副轮状病毒(C群),且可引起仔猪爆发流行,是仔猪腹泻不可忽视的病原之一。     

B组轮状病毒RT—PCR来源的cDNA探针核酸诊断方法的建立

近十年来,国内外学者相继报道在人以及牛、羊、猪、大鼠的腹泻粪样中检出Ｂ组轮状病毒〔１－３〕。洪涛等于１９８３年首次报道成人腹泻轮状病毒（ＡＤＲＶ）属于Ｂ组轮状病毒〔４〕。目前,轮状病毒Ｂ组已被公认为引起人及不同动物腹泻的流行病学的重要因素。由于Ｂ组轮...     

人B组轮状病毒vp7基因的克隆和序列分析

利用逆转录—聚合酶链反应(RT-PCR)技术,扩增了人B组轮状病毒WH—2vp7基因,连接到克隆载体pUCm—T,并对其基因序列进行了分析。WH—2与ADRV的同源性达98％,与印度加尔各达分离株CAL—1达92％,而与动物B组轮状病毒同源性差别较大,如与IDIR(鼠)同源性仅为58％,与WD653(牛)B组轮状病毒同源性为63％,与ATI(牛)同源性为61％。对vp7基因的二级结构分析发现其mRNA折叠形成多达18个发卡环状结构。VP7蛋白是249个氨基酸组成,分子：量为28．4kDa,含有3个潜在的N连接的糖基化位点和多个磷酰化位点,从氨基酸序列的同源性来看,WH—2与ADRV的同源性达99％,与CAL—1达95％,而IDIR仅为51％,说明了WH—2和ADRV的起源相同。此研究对了解B组轮状病毒基因的进化和变异规律具有重要意义,也将为B组轮状病毒预防性疫苗的研制提供科学的依据。     

成人腹泻轮状病毒蛋白的免疫印迹分析

我们从腹泻病人便样中纯化了病毒,其结构蛋白组分按分子量大小分别为VP1(136K),VP2(113K),VP3(92K),VP4(84K),VP5(64K),VP6(47K),VP7(41K)。所有这些蛋白皆具有抗原性。WP6是B组轮状病毒的共同抗原。B组轮状病毒的每一结构蛋白与A组轮状病毒都无交叉免疫反应。另外注意到二例不同病人对VP6和VP7刺激产生的抗体水平不同。     

Adult diarrhea rotavirus in Jilin, China

In May 1986, an outbreak of epidemic acute diarrhea occurred in one city and three counties in Yanbian area. The diarrheal cases were seen in all age groups, the majority of the cases were seen in adult age group. No bacterial pathogens were isolated in 22 fecal samples examined, however rotavirus like particles of 52-68 nm in diameter were found in 50% (11/22) of fecal samples by the immunoelectron microscopy using convalescent sera. Examination by the adult diarrhea rotavirus (ADRV) ELISA kit showed positive reaction in 86% (6/7) of fecal extracts, however in all 22 fecal extracts examination using ELISA kit for detection of group A rotavirus showed negative reaction. The PAGE patterns of viral RNAs similar to those of ADRV were seen in 13 out of 22 fecal extracts. The detection rate by PAGE analysis was 59%. Based on the above data, the etiological agent of the epidemic acute diarrhea, which occurred in Yanbian area, was identified to be ADRV.     

Purification and characterization of adult diarrhea rotavirus: identification of viral structural proteins.

Adult diarrhea rotavirus (ADRV) is a newly identified strain of noncultivable human group B rotavirus that has been epidemic in the People's Republic of China since 1982. We have used sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western (immuno-) blot analysis to examine the viral proteins present in the outer and inner capsids of ADRV and compared these with the proteins of a group A rotavirus, SA11. EDTA treatment of double-shelled virions removed the outer capsid and resulted in the loss of three polypeptides of 64, 61, and 41, kilodaltons (kDa). Endo-beta-N-acetylglucosaminidase H digestion of double-shelled virions identified the 41-kDa polypeptide as a glycoprotein. CaCl2 treatment of single-shelled particles removed the inner capsid and resulted in the loss of one polypeptide with a molecular mass of 47 kDa. The remaining core particle had two major structural proteins of 136 and 113 kDa. All of the proteins visualized on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were antigenic by Western blot analysis when probed with convalescent-phase human and animal antisera. A 47-kDa polypeptide was most abundant and was strongly immunoreactive with human sera, animal sera raised against ADRV and against other group B animal rotaviruses (infectious diarrhea of infant rat virus, bovine and porcine group B rotavirus, and bovine enteric syncytial virus) and a monoclonal antibody prepared against infectious diarrhea of infant rat virus. This 47-kDa inner capsid polypeptide contains a common group B antigen and is similar to the VP6 of the group A rotaviruses. Human convalescent-phase sera also responded to a 41-kDa polypeptide of the outer capsid that seems similar to the VP7 of group A rotavirus. Other polypeptides have been given tentative designations on the basis of similarities to the control preparation of SA11, including a 136-kDa polypeptide designated VP1, a 113-kDa polypeptide designated VP2, 64- and 61-kDa polypeptides designated VP5 and VP5a, and several proteins in the 110- to 72-kDa range that may be VP3, VP4, or related proteins. The lack of cross-reactivity on Western blots between antisera to group A versus group B rotaviruses confirmed that these viruses are antigenically quite distinct.     

Epidemiology of leptospirosis: observations on serological data obtained by a "diagnostic laboratory for leptospirosis" from 1995 to 2001

Serological data on leptospira infection were reported and discussed. From 1995 to 2001, the blood serum samples of 9885 domestic and wild animals and humans, living in Northern and Central Italy, were examined by the macroagglutination test (MAT) employing bratislava, ballum, canicola, grippotyphosa, icterohaemorrhagiae, pomona, hardjo and tarassovi serovars as antigens. Considering sera with > or = 1:400 antibody titers as positive, 674 (6.81%) animals scored positive. Sheep, horses, pigs and dogs gave the highest number of positive responses, particularly against the serovar bratislava and, for dogs, against icterohaemorrhagiae. The percentages of seropositivity observed in the most important animal species were: 12.13% in ovine (132 positive among 1088 tested animals), 11.40% in horses (107 positive animals among 938), 9.46% in swine (123 positive animals among 1299), 6.36% in dogs (278 positive animals among 4369), 2.39% in wild boars (11 positive animals among 459), 1.39% in deer (2 positive animals among 143), 0.48% in cattle (3 positive animals among 626). Among 250 human sera examined, 14 (5.60%) scored positive.     

Superficial infections caused by Microsporum canis in humans and animals

Dermatophytic infections caused by M. canis in humans and animals have a world wide distribution and they are zoonotic. The objective in this work was to know the frequency of M. canis infections in humans and pets. We studied our cases from January 1994 to December 2002. The human samples were obtained from a Dermatological Department in a General Hospital and we registered the next data: age, sex, job, and affected area. The animal samples were obtained from a mycological veterinary laboratory, and we registered the presence or absence of clinical lesions. A total of 46 clinical cases of M. canis infections were recorded, 26 female and 20 males: tinea capitis 21, tinea corporis 17, tinea pedis five, onychomycosis two, and only one case with tinea faciei. The 46 cases with positive culture yield 42 positive samples in KOH. The age range varied from 2 to 60 years. Among the animals, we studied 461 dogs and found six KOH positive (1%) samples and cultured 23 isolates (4.98%): 21 M. canis, one M. gypseum and one Trichophyton spp. From the 68 samples of cats, eight (11.76%) were positive to KOH, being 26 (38.23%) M. canis isolates. In M. canis infections in humans, the age rage was wide with predominance in women. In animals, M. canis isolates represented the most dermatophytic infection.