肠道传播的非甲非乙型肝炎病人粪便排病毒规律的研究

1986～1987年新疆南部和田发生了一起肠道传播的非甲非乙型肝炎的流行。本研究用免疫电镜方法(IEM)对6例急性期病人的60份粪便标本进行了排病毒规律的检测。其中5例排病毒阳性(83.3％);全部粪便标本的病毒颗粒检出率为28.3％(17/60)。发病前1～4天的粪便标本阳性率为100％(3/3);发病后9～12天的阳性率为14.3％(1/7);发病两周后的22份标本全部阴性。17份不同病期收集的阳性粪便标本,94.1％(16/17)出现在病人血清转氨酶(SGPT)高峰值前,此时SGPT在100IU/L以下。故此,肠道传播的非甲非乙型肝炎患者的隔离期,应定为病后2～3周。     

戊型肝炎研究进展

戊型肝炎既往称为肠道传播的非甲非乙型肝炎。世界上首次有记载的本病流行系发生于1955～1956年新德里,共计发病97000例,其中29300例为黄疽型肝炎。1980年Khuroo等报告,1978年11月～1979年4月克什米尔流域发生本病流行。1983年Balayan等首次用成型肝炎病人粪便经口感染一名志愿者获得成功,并从急性期粪便中用免疫电镜检测到     

ELISA检测椰酵假单胞菌菌体抗原的研究

椰毒假单胞菌酵米面亚种,已知有0—Ⅲ、0—Ⅳ和0—Ⅴ等菌体抗原型。常规血清试管凝集法检测分型耗时费力。本文在制备不同血清型菌体抗原的抗体和辣根过氧化物酶抗体结合物的基础上,建立了检测椰酵假单胞菌的间接非竞争法、直接非竞争法和双抗体夹心ELISA法,具有快速、简便和较好的特异性。     

枭猴和猕猴戊型肝炎病毒的实验感染

戊型肝炎通常以猕猴作为动物实验模型,但枭猴能否作为戊型肝炎的动物模型尚缺乏报告.为此作者进行了此项研究.实验用戊型肝炎病毒(HEV)取自墨西哥1986年肠道传播的非甲非乙型肝炎(ET--NANBH)病人粪便中提取的Telixtac一14毒株。     

从病毒性肝炎病人血清中检测到的一种沉淀抗原的特点

作者在探索非甲非乙型肝炎(HNANB)的病原过程中,用HNANB病人的恢复期血清和血友病患者的血清作为抗体,应用琼脂扩散法从HNANB急性期病人的血清中,检测到一种沉淀抗原。本抗原于HNANB急性期血清中出现,发病后1～5周(平均3周)消失,恢复期血清中出现相应抗体。该抗体于出现尿黄后3周才能检测到,可持续1～10周。抗原阳性血清用相应抗体中和     

诊断丙型肝炎病毒感染的3种ELISA的比较研究

将新建立的检测血清中抗丙型肝炎病毒（ＨＣＶ）总抗体的双抗原夹心ＥＬＩＳＡ（夹心法）与普遍采用的检测抗ＨＣＶ－ＩｇＧ的间接ＥＬＩＳＡ（ＩｇＧ间接法）及检测抗ＨＣＶ－ＩｇＭ的间接ＥＬＩＳＡ（ＩｇＭ间接法）进行比较研究。检测１５份ＩｇＭ间接法阳性、ＩｇＧ间接法阴性（ＩｇＭ单阳）血清及２０份ＩｇＧ间接法阳性、ＩｇＭ间接法阴性（ＩｇＧ单阳）血清,夹心法均为阳性,这３５份血清经ＰＣＲ检测ＨＣＶＲＮＡ均阳性。夹心法可同时检出ＩｇＧ和ＩｇＭ抗体,检出率高于后两者。检测２０份ＨＣＶＲＮＡ阴性血清,３种方法均阴性。用生物制品检定所检测ＨＣＶ－ＩｇＧ抗体的质控血清为标准品,夹心法和ＩｇＧ间接法均可达到检测标准     

双抗原夹心法ELISA在肾综合征出血热诊断中的应用

目的建立一种敏感、特异的检测肾综合征出血热(HFRS)血清总抗体的双抗原夹心法ELISA。方法应用重组汉坦病毒(HV)NP抗原e1.3S作为包被抗原和e6-119-HRP作为酶标抗原建立双抗原夹心法ELISA,检测血清总抗体,并与间接免疫荧光法(IFA)相比较。结果共检测188份人血清和378份鼠血清标本,2种方法的总符合率为97.70%。以IFA为参照标准,双抗原夹心法ELISA的敏感性为97.54%,特异性为97.87%。结论双抗原夹心法ELISA检测HFRS血清总抗体具有很高的敏感性和特异性,适用于大规模的流行病学调查。     

抗副溶血弧菌TLH蛋白多克隆抗体的制备及其ELISA双抗体夹心检测法的研究

目的分别制备抗副溶血弧菌及其不耐热溶血毒素(thermolabile hemolysin,TLH)的多克隆抗体,建立检测副溶血弧菌TLH的酶联免疫吸附试验(ELISA)双抗体夹心法。方法以副溶血弧菌及其TLH分别免疫健康雄性新西兰大白兔和雌性BALB/C小鼠,制备兔抗副溶血弧菌多克隆抗体及小鼠抗TLH多克隆抗体,以间接ELISA法检测两种抗体效价,并以棋盘法筛选两种多克隆抗体用于夹心检测TLH的效价。结果兔抗副溶血弧菌多克隆抗体效价达1∶6400,鼠抗TLH多克隆抗体效价为1∶102 400,经筛选确定以1∶800稀释的兔抗副溶血弧菌多克隆抗体包被酶标板,与1∶1600稀释的鼠抗TLH抗体建立了ELISA双抗体夹心法。结论兔抗副溶血弧菌多克隆抗体可与TLH呈特异性反应,以此建立的双抗体夹心法检测副溶血弧菌TLH可获较满意结果,此实验可为进一步建立简便、快速的副溶血弧菌检测方法奠定基础。     

聚合酶链反应标记输血传播病毒（TTV）地高辛霉探针的初步应用

用聚合酶链反应法（ＰＣＲ）制备地高辛素标记的输血传播病毒（ＴＴＶ）探针,并与巢式ＰＣＲ法比较。结果 该探针具有ＴＴＶ特异性,其灵敏度为１０ｐｇ ＤＮＡ。应用该法检测１０８份非甲￣庚型肝炎病人的血清标本,应用该法检测１０８份非甲￣庚型肝炎病人的血清标本,ＴＴＶ ＤＮＡ阳性率为１８．５％（２０／１０８）;检测２２份病人的粪便标本,ＴＴＶ ＤＮＡ阳性率为２７．３％（６／２２）。该法与ｎＰＣＲ法的总符合率     

对消化系疾病血清不溶性白细胞介素2受体的探讨

对消化系疾病血清不溶性白细胞介素２受体的探讨大连市中医医院（１１６０１３）王浩明大连大学医学院（１１６０２１）董正义我们在实践中用双抗体夹心酶联免疫吸附（ＥＬＩＳＡ）法对１７８例常见各种消化系疾病病人进行了血清ＳＩＬ－２Ｒ检测，认为血清ＳＩＬ－２Ｒ可...     

Use of enzyme linked immunosorbent assay in intestinal and extra-intestinal amoebiasis (amoebic liver abscess)

The sera of patients with amoebic liver abscess, intestinal amoebiasis and non-specific hepatomegaly were examined for the presence of antibodies to Entamoeba histolytica antigen by enzyme linked immunosorbent assay (ELISA). All the amoebic liver abscess sera were strongly positive. The test was also positive in all the proved cases of intestinal amoebiasis but the ELISA values were relatively slightly lower. Nine of 10 cases of non-specific hepatomegaly with stools negative for E. histolytica gave positive results with ELISA. The IHA titre in all but one of the non-specific hepatomegaly cases was within normal value.     

Identification of human and bovine rotavirus serotypes by polymerase chain reaction.

The use of the polymerase chain reaction (PCR) for identifying serotypes of human and bovine rotaviruses was examined. In the identification of 115 human rotavirus samples in stools, results with PCR showed excellent agreement with results of serotyping by an enzyme-linked immunosorbent assay (ELISA) using serotype-specific monoclonal antibodies. Furthermore, the PCR showed a much higher sensitivity (93%) than the ELISA test (82.6%). The PCR method could also be applied for identifying the serotype of bovine rotaviruses.     

Detection of Clostridium difficile toxins by enzyme immunoassay

An enzyme-linked immunosorbent assay (ELISA) for the rapid diagnosis of antibiotic-associated colitis (AAC) is presented. Commercially available antisera to Clostridium difficile toxins contain antibodies to other antigens found in non-toxigenic C. difficile and other bacteria. Removal of these unwanted antibodies by absorption increased the specificity of ELISA for detection of C. difficile toxins. Specimens tested included 40 faecal extracts positive for cytotoxicity from cases of AAC, 30 diarrhoeic and 30 well-formed stools negative for cytotoxicity and 50 culture filtrates of toxigenic and non-toxigenic C. difficile and other clostridial species. Use of absorbed sera reduced false-positive reactions observed with faecal specimens from 23 to 8%. About 90% of specimens that were positive by the tissue culture cytotoxicity test were positive by ELISA using the absorbed sera. The relative merits of ELISA and other methods for the rapid diagnosis of AAC are discussed.     

Detection of Clostridium difficile toxins by enzyme immunoassay.

An enzyme-linked immunosorbent assay (ELISA) for the rapid diagnosis of antibiotic-associated colitis (AAC) is presented. Commercially available antisera to Clostridium difficile toxins contain antibodies to other antigens found in non-toxigenic C. difficile and other bacteria. Removal of these unwanted antibodies by absorption increased the specificity of ELISA for detection of C. difficile toxins. Specimens tested included 40 faecal extracts positive for cytotoxicity from cases of AAC, 30 diarrhoeic and 30 well-formed stools negative for cytotoxicity and 50 culture filtrates of toxigenic and non-toxigenic C. difficile and other clostridial species. Use of absorbed sera reduced false-positive reactions observed with faecal specimens from 23 to 8%. About 90% of specimens that were positive by the tissue culture cytotoxicity test were positive by ELISA using the absorbed sera. The relative merits of ELISA and other methods for the rapid diagnosis of AAC are discussed.     

Prevalence of enteric adenovirus from acute gastroenteritis: A five year study

Adenoviruses were detected in the stools in 459 of 3932 (111.6%) pediatric children hospitalized with acute gastroenteritis from January 1981 to December 1985. Out of the 459 adenovirus specimens 325 (8.3%) were presumptively identified as enteric adenovirus both by an adenovirus genus specific ELISA and by growth characteristics of adenovirus isolates in HEp-2 cells and in 293 cells. Enteral adenoviruses were found endemic since these viruses have been found for 5 successive years. A seasonal variation in the rates of adenovirus was also observed. Comparative data of rotavirus prevalence in the same study population are reported.Corresponding author.     

Solitary IgM phase II response has a limited predictive value in the diagnosis of acute Q fever

SUMMARY We investigated the positive predictive value (PPV) of a solitary positive immunoglobulin M (IgM) phase II response for the serodiagnosis of acute Q fever detected with either an indirect immunofluorescence assay (IFA) or an enzyme-linked immunosorbent assay (ELISA). Initial and follow-up sera from patients suspected of acute Q fever were included if initially only IgM phase II tested positive with IFA in 2008 (n=92), or ELISA in 2009 (n=85). A seroconversion for Q fever was defined as an initial sample being IgG phase II negative but positive in the follow-up sample. The PPV of an initial isolated IgM phase II result detected by IFA or ELISA was 65% and 51%, respectively, and therefore appeared not to adequately predict acute Q fever. For this reason it cannot be used as a diagnostic criterion nor should it be included in public health notification without confirmation with other markers or a follow-up serum sample.     

TaqMan实时荧光定量PCR检测肠道细菌Akkermansia Muciniphila

目的 建立Akkermansia muciniphila(A.muciniphila)TaqMan实时荧光定量PCR检测方法,实现粪便中A.muciniphila的定量检测。方法 根据GenBank公布的A.muciniphila 16S rRNA基因高保守序列,设计合成特异引物和TaqMan探针,构建重组质粒作为标准品,通过反应体系和反应条件的优化,绘制标准曲线,同时将建立的方法用于10份成年人粪便标本的检测。结果 TaqMan实时荧光定量PCR检测A.muciniphila,仅需1.5h,该方法线性好,相关系数R²=0.9991,扩增效率E=94.7%;重复性好,组内和组间变异系数均小于3%;灵敏度可达到5×10³拷贝/μl;特异性良好。10份成年人粪便标本共检出9份为阳性,阳性粪便标本中A.muciniphila平均含量为2.70×108cells/g粪便。结论 本研究建立的TaqMan实时荧光定量PCR能够快速、灵敏、特异地定量检测粪便标本中A.muciniphila。     

北京市某医院院内感染Noro病毒胃肠炎的调查

目的了解Noro病毒感染引起医院内暴发集体腹泻的临床特点。方法分析一起18例医院内暴发腹泻患者的流行病学和临床资料,同时对每例患者粪便标本进行细菌培养,随机选择7例粪便标本用抗原检测和聚丙烯酰胺凝胶电泳进行轮状病毒RNA检测;用逆转录-聚合酶链反应检测Noro病毒核酸以确定感染的病原,分析感染患者的临床特点和易感因素。结果发病者多为老年合并多种基础疾病;腹泻主要为稀水便(12/18,66.67%),个别为黏液稀便(3/18,16.67%);大部分粪便常规检测正常(10/18,55.56%),个别见少量白细胞(3/18,16.67%)和潜血弱阳性(4/18,22.22%);全部粪便标本细菌学培养阴性;轮状病毒RNA检测阴性;3份标本中检出Noro病毒核酸,阳性率为42.86%(3/7)。结论Noro病毒是此次医院内集体暴发腹泻的重要病原之一,尤其多见于有基础疾病、体弱的老年患者。     

Enterotoxigenic Escherichia coli in the domestic environment of a Malaysian village

The membrane-filter assay, GM1-ELISA, and DNA-DNA hybridization assay, were used to detect enterotoxigenic Escherichia coli (ETEC) in samples of water, weaning food, food preparation surface swabs, fingerprints of mothers, and the fingerprints and stools of children under 5 years of age, in 20 households in a Malaysian village. Weaning food and environmental samples were frequently contaminated by faecal coliforms, including ETEC. The membrane-filter assay detected and enumerated faecal coliforms and LT-ETEC in all types of water and weaning food samples. Highest concentrations of faecal coliforms and LT-ETEC were found in weaning food, followed by well-water, stored water and stored drinking water. The GM1-ELISA detected LT-ETEC in weaning food, food preparation surfaces, fingerprints and stool samples. The DNA-DNA hybridization assay detected a larger proportion of STa2-ETEC than the other toxotypes, either singly or in combination. All the assays in combination detected the presence of ETEC in all types of samples on at least one occasion in each household. It was not possible to classify households as consistently more or less contaminated with ETEC. On individual occasions it was possible to show a significant association of the presence of LT-ETEC between the fingerprints of children and their stools, fingerprints of mothers and children, and weaning food and the stools of the child consuming the food.     

Diarrheal diseases in Brazil: Clinical features of rotavirus-associated gastroenteritis in children

This study was undertaken to detect the presence of rotavirus in the stools of children with gastroenteritis, using the enzyme-linked-immunosorben assay (ELISA), and to compare the signs and symptoms of rotavirus-positive and -negative children. Over a period of fifteen months, 367 children ranging in age from less than 1 month to 5 years or more with diarrhea and 86 children, in the same age group, without diarrhea and respiratory infections, used as controls, were evaluated. Human rotavirus was detected in 15.8% of children with diarrhea attending out-patient clinics and in 28.9% of patients seen by general practitioners. In the control groups, the percentages of identification of rotavirus were 1.4% and 5.5% respectively. Frequency of other enteropathogens was determined. The hydration state of diarrheal cases, different clinical symptoms and the type of medical attendance distinguished the rotavirus positive from the rotavirus negative patients.