腹泻患儿粪便A群轮状病毒抗原检测的结果分析

目的:了解本地区腹泻婴幼儿的A群轮状病毒感染情况及其流行特点。方法:采用胶体金法对我院2009年10月-2010年9月2104例有腹泻和肠炎特征的婴幼儿粪便进行A群轮状病毒抗原检测。结果:在2104例受检者中,A群轮状病毒感染的总阳性率是24.71%,其中男性感染率24.17%,女性为25.40%。不同年龄组间以1-2岁婴幼儿感染率最高,为32.13%,0-1岁为20.72%,2-5岁为12.03%。感染的季节特征是秋末冬初季(10-12月)阳性率最高,为42.82%,春末夏初季(4-6月)最低,为8.81%。结论:由A群轮状病毒感染引发的急性腹泻主要发生在1-2岁的婴幼儿,各个季节均有发生,以秋末冬初季为高发。     

腹泻患儿粪便中A群轮状病毒抗原阳性率及其相关因素分析

目的回顾性分析我院腹泻患儿粪便中A群轮状病毒抗原检测情况,为临床提供快捷、准确、可靠的诊断依据。方法收集腹泻患儿粪便标本2 869例,应用免疫层析双抗体夹心法检测A群轮状病毒抗原,对阳性率及其影响因素进行统计分析。结果2 869例检测结果中阳性946例,总阳性率为32.9%。1~2岁患儿阳性率最高,为36.5%;0~1岁阳性率其次,为32.3%;2~6岁患儿阳性率相对较低,为27.0%。不同年龄段间比较差异具有统计学意义(χ2=11.17,P0.05)。秋冬季节为轮状病毒感染率的高峰期,春夏季节较低,分别为39.5%和22.6%。不同季节两组间比较差异具有统计学意义(χ2=10.15,P0.05)。并且发现粪便以黏液便或水样便为特征的患儿A群轮状病毒感染率较高。结论轮状病毒是婴幼儿急性重症腹泻的主要病原体之一,及时进行轮状病毒抗原检测,对腹泻患儿的及时诊断和合理预防治疗具有重要意义。     

轮状病毒腹泻患儿检测结果分析

目的:探讨轮状病毒感染与天气因素之间的关系,以指导临床采取合理的防治策略.方法:采用免疫胶体金法对1155例腹泻患儿进行轮状病毒检测;收集2005年到2007年的天气资料;分别分析轮状病毒阳性患儿人次与温度、湿度之间变化的规律.结果:501份标本轮状病毒阳性,6m-24m年龄段婴幼儿阳性检出率最高.每年秋季(第33周时)随着周平均气温缓慢下降患儿人次开始逐渐上升,到第41周患儿人数达到最高峰,之后回落,达高峰前几周湿度也略有下降,表明天气因素与轮状病毒的传播有密切的关系.结论:秋季周平均气温、相对湿度是影响轮状病毒传播流行的自然因素.     

婴幼儿腹泻轮状病毒和腺病毒检测结果分析

目的了解本地区婴幼儿腹泻患者中轮状病毒和腺病毒的感染情况,为临床治疗提供依据。方法收集2013年1月-12月门诊和住院腹泻患儿的粪便,进行轮状病毒和腺病毒抗原检测,并对结果进行统计学分析。结果在检测的2 579例婴幼儿腹泻中轮状病毒阳性为616例,阳性率为23.89%,高发季节为1月、11月和12月,高发年龄组为13-24月婴幼儿,腺病毒阳性共102例,阳性率为3.96%,程散发状况,高发年龄组为7-12月的婴幼儿。结论轮状病毒和腺病毒都能引起婴幼儿腹泻,但是轮状病毒为最主要的病原体,及时检测轮状病毒抗原,为临床治疗和疾病监测提供参考。     

婴幼儿腹泻A群轮状病毒G和P的基因分型研究

目的研究浙江萧山医院婴幼儿童腹泻标本中人轮状病毒（Human Rotavims）毒株的感染情况及G和P基因型流行特点。方法收集该院2009年8月至2010年8月腹泻儿童15233份粪便标本采用酶联免疫吸附试验、逆转录-巢式聚合酶链反应进行轮状病毒病原检测,将128份阳性标本进行VP7和VP4基本分型。结果15233份婴幼儿腹泻标本中有2706份标本为轮状病毒阳性,阳性率17．8％;男孩和女孩检出率差异无统计学意义,以6-12月龄段检出率最高;对128份阳性标本进行G血清分型和P基因分型,G1型53份（41．4％）、G3型38份（29．7％）、G1G3型17份（13．3％）、G未分型20份（15．6％）;P[8]型72份（56．3％）、P[4]型16份（12．5％）、P[8]P[4]型3份（2．3％）、P未分型37份（28．9％）,G血清型和P基因型的组合以G1P[8]为主,占29．7％（38／128）。结论浙江萧山医院A群轮状病毒G血清以G1型为主,其次为G3型,P基因型以P[8]型为主。     

膜芯片检测A组轮状病毒的初步研究

建立表面带正电荷的尼龙膜为基片的基因芯片,采用RT-semi-nested PCR方法,对A组轮状病毒RNA实现高度扩增,并通过5'端带地高辛标记的上游引物实现扩增产物的标记.通过同膜芯片上探针的杂交和免疫显色,实现对A组轮状病毒的检测.结果表明,膜芯片对探针的固定效果、杂交吸脱和检测结果可靠,空白对照和阴性对照均为阴性,探针均显示为阳性信号,并且信号强弱与探针浓度关系不大,而主要与探针本身同互补链的结合相关.以上结果说明已经初步建立了快速检测A组轮状病毒的膜基因芯片检测技术.     

三种检测婴幼儿腹泻A组轮状病毒的方法学评价

目的对婴幼儿腹泻A组轮状病毒的三种不同检测方法进行评价。方法同时采用胶体金、琼脂糖凝胶电泳、逆转录PCR（RT-PCR）对浙江萧山医院1周320份腹泻患儿的标本进行轮状病毒检测,然后进行方法学比较。结果三种方法的阳性检出率分别为36．3％、48．1％和42．8％,经卡方检验三种方法的检出率之间差异均存在统计学意义（P〈0．05）;以琼脂糖凝胶电泳为检测的最终标准,RT-PCR的敏感性为88．9％,特异性达100％;胶体金法的敏感性71．4％,特异性达96．4％。结论胶体金法检测轮状病毒的敏感性和特异性能够满足临床上对轮状病毒快速筛检的要求;PT-PCR法的敏感性和特异性均最高,既可用于临床检测,又是轮状病毒血清G、基因P分型的重要手段,在对无菌体液和环境样本中轮状病毒的检测具有不可比拟的优势。     

沙市婴幼儿腹泻中检出3株C群轮状病毒

1987～1988年在沙市165份婴幼儿急性腹泻标本中,用PAGE法检出轮状病毒37株(22.4％),其中3株为少见的轮状病毒,此种病毒经电镜观察,具有典型轮状病毒的形态结构,ELISA证实该病毒不具有A群和B群轮状病毒的群特异性抗原。RNA电泳分析表明,其基因组由11个双链RNA片段组成,电泳图型特殊,呈4:3:2:2的排列模式。上述试验表明,该病毒为世界上罕见的C群轮状病毒。免疫电镜证实,该病毒能被病人恢复期血清所凝集,提示该病毒是腹泻病儿的致病因子。     

两种PCR方法对检测A组轮状病毒膜芯片杂交结果的影响

A组轮状病毒(rotavirus,RV)是导致拿世界婴幼儿腹泻的最主要病原,危害巨大。拟用RT-巢式PCR技术对A组RV的保守序列进行高度扩增,通过固本室内制的膜芯片杂交,实现对该病毒的检测。分别采用对称PCR和不对称PCR扩增,均可得到扩增的目的片段．对称式扩增产物杂交结果不理想。而不对称式扩增得到了大量待检单链产物,同膜芯片杂交获得了理想的杂交结果。显著地提高了对A组RV杂交检测的灵敏度。表明不对称式PCR扩增是一种制备用于芯片杂交大量单链产物的理想方法,尤其是针对富含AT的核酸检测区域。     

轮状病毒感染成年小鼠的研究

目的研究成年昆明种小鼠对实验感染人轮状病毒(rotavirus,RV)的敏感性。方法在实验条件下,用A组人Wa和恒河猴SA11株RV感染成年昆明种小鼠,观察小鼠的临床反应和排毒情况。结果成年昆明种小鼠感染Wa和SA11RV第二天后出现明显的临床腹泻症状,第四天达到高峰;至少在感染后连续6天的动物大便中可检测到较高滴度的RV抗原。结论成年昆明种小鼠对RV感染有很高的敏感性,可做为动物模型,在RV感染的药物治疗效果评价和疫苗保护性效果评价中具有重要价值。     

A组人轮状病毒全基因组克隆和基因型分析

运用基因克隆和重组技术,从一急性胃肠炎患儿样品中克隆到一株轮状病毒（RV）全基因组cDNA。核苷酸序列测定结果表明,该株RV基因组11条RNA共含有18613个核苷酸（3302bp—751bp）,编码5791个氨基酸。全基因组研究结果表明,该株RV（TB-Chen）为A组,II亚组,基因型为G2P[4]/NSP4[A]。这是首个A组人RV全基因组研究结果的报道,对研究和开发RV疫苗具有积极的意义。     

Rotavirus vp7 antigen produced by Lactococcus lactis induces neutralizing antibodies in mice

AIMS: To determine if live recombinant Lactococcus lactis strains expressing rotavirus VP7 antigen are immunogenic in mice. METHODS AND RESULTS: Using the food-grade lactic acid bacterium L. lactis as a carrier, we expressed VP7, the major rotavirus outer shell protein and one of the main components of the infective particle, as a cytoplasmic, secreted or cell wall anchored forms. Our results showed that recombinant L. lactis strains secreting VP7 proved to be more immunogenic than strains containing the antigen in the cytoplasm or anchored to the cell wall. CONCLUSIONS: This is the first demonstration that recombinant L. lactis producing VP7 can induce the production of a neutralizing antibody response against rotavirus by the intragastric route. SIGNIFICANCE AND IMPACT OF THE STUDY: Rotaviruses are the single most important aetiological agents of severe diarrhoea of infants and young children worldwide and have been estimated to be responsible for 650 000-800 000 deaths per year of children younger than 5 years old in development countries. Thus, the development of a safe and effective vaccine has been a global public health goal. Although two of five mice orally inoculated with L. lactis strains secreting VP7 elicited a specific-antibody response, these strains could be very useful to be used as a prototype to develop a new generation of protective rotavirus vaccines.     

参苓白术散联合推拿治疗急性轮状病毒感染性腹泻患儿的疗效及对胃肠激素和免疫力的影响

摘要 目的：探讨参苓白术散联合推拿治疗急性轮状病毒感染性腹泻患儿的疗效及对胃肠激素和免疫力的影响。方法：选取2018年12月~2019年12月期间我院收治的80例急性轮状病毒感染性腹泻患儿,按照随机数字表法分为对照组（n=40）和研究组（n=40）,对照组患儿予以常规基础治疗,研究组在对照组基础上给予参苓白术散联合推拿治疗,比较两组患儿疗效、中医证候积分、胃肠激素指标[胃泌素（GAS）、胃动素（MOT）以及血管活性肠肽（VIP）]和免疫力指标[免疫球蛋白 A（IgA）、免疫球蛋白 G（IgG）、CD4⁺ /CD8⁺],记录两组不良反应发生情况。结果：研究组治疗7 d后的临床总有效率为95.00%（38/40）,高于对照组的67.50%（27/40）（P<0.05）。两组治疗7 d后GAS、MOT、VIP均下降,且研究组低于对照组（P<0.05）。两组治疗7 d后IgA、IgG、CD4⁺ /CD8⁺均升高,且研究组高于对照组（P<0.05）。两组治疗期间不良反应发生率对比未见统计学差异（P>0.05）。两组治疗7 d后腹痛、大便次数、大便性状、体温中医证候积分均降低,且研究组低于对照组（P<0.05）。结论：参苓白术散联合推拿治疗急性轮状病毒感染性腹泻患儿,疗效显著,可有效改善患儿胃肠状况,提高患儿免疫力,且安全性较好。     

Novel monoclonal antibody-based Helicobacter pylori stool antigen test

BACKGROUND: A number of noninvasive tests have been developed to establish the presence of Helicobacter pylori infection. Although polyclonal antibody-based stool antigen testing has a good sensitivity and specificity, it is less accurate than urea breath testing. Recently, a monoclonal antibody-based stool antigen test demonstrated an excellent performance in diagnosing H. pylori infection in adults and in pediatric populations. AIM: To evaluate the diagnostic accuracy of a novel stool test based on monoclonal antibodies to detect H. pylori antigens in frozen human stool in the pretreatment setting. PATIENTS AND METHODS: Stool specimens were prospectively collected from 78 patients undergoing gastroscopy and stored at -20 degrees C until tested. Helicobacter pylori infection was evaluated by histology, rapid urease testing and urea breath tests ((13)C-UBT). Positivity of the three tests was considered the gold standard for H. pylori active infection. Patients with no positive test were considered negative. The gold standard was compare to the results of the monoclonal antibody stool antigen test. Frozen stool specimens were tested using a novel monoclonal-antibody-based enzyme immunoassay (HePy-Stool, Biolife-Italiana, Milan, Italy). RESULTS: The sensitivity and specificity of the monoclonal stool antigen test were 97%[95% confidence interval, (CI) 86-100] and 94% (95% CI: 81-99), respectively. Negative and positive predictive values were 97% (95% CI: 85-99), and 95% (95% CI: 83-99), respectively. The diagnostic accuracy was 96% (95% CI: 88-99). The likelihood ratio for a positive test was 17 and for a negative test was 0. CONCLUSIONS: Although the (13)C-UBT is the most accurate among the available noninvasive tests, our results show that an H. pylori stool test using monoclonal antibody might be an excellent alternative.     

Detection of human rotavirus in hospitalized diarrheic children in central India

During the present study, group A human rotaviruses were detected among diarrheic children using polyacrylamide gel electrophoresis (PAGE) technique, with a typical RNA migration pattern of 4:2:3:2, suggestive of group A rotavirus. During the study, a total of 46 fecal samples collected from hospitalized children with acute diarrhea as well as children inhabiting nearby animal farms with history of presence of animal rotaviruses on the farms were processed for detection of human rotavirus. Out of 33 diarrheic children, 12 showed presence of rotavirus infection (36.36%), however, none of the children from animal farm areas showed presence of rotavirus. Female children were more susceptible to rotavirus infection (46.15%) than males (30%). Majority of the cases of rotavirus gastroenteritis belonged up to one year of the age, with an incidence of 40.91%. RNA profile of rotaviruses suggested circulation of 5 different electropherotypes in this geographical locale of the country, indicating existence of genomic diversity among human rotaviruses. Majority of the isolates were of long pattern (66.67%), whereas short pattern was detected only in one third of the viruses. This preliminary study emphasizes for further detailed studies on the molecular characterization of rotaviruses circulating in this part of country and their relationship with other human rotavirus strains and animal strains in the country.