广州地区婴幼儿急性下呼吸道感染的病原学研究

1973年12月至1986年7月的13年间,我们对住院诊断为婴幼儿急性下呼吸道感染病例的病原学进行了研究,分析广州地区本病流行的特点。材料和方法一、材料对1973年至1986年5月在本院诊断为急性下呼吸道感染的婴幼儿,采取鼻咽(分泌物)拭子1600份,其中131份为气管分泌物,和急性期及恢复期血清1055份。     

肠道病毒71型VP1基因的扩增、克隆、表达及蛋白分析

目的 对肠道病毒71型VP1基因片段进行扩增、克隆、生物信息学分析、原核表达、纯化,初步证实重组表达产物的生物学活性。方法 根据GenBank中EV71序列设计一对特异性引物,以EV71患者咽拭子标本中提取病毒核糖核酸为模板,RT-PCR扩增EV71 VP1基因,酶切后插入表达载体pET28a。构建pET28a-EV71 VP1原核表达载体。转化E.coli DH5α,IPTG诱导表达,使用SDS-PAGE及Western blot分析表达结果,利用软件对测序结果进行生物信息学分析。纯化蛋白并包被反应板,用ELISA分别检测EV71阳性与COX A16阳性患者VP-1IgG抗体,进行统计学分析。结果 目的基因经BLAST比对,同源性与GenBank登录号为JQ766207.1 EV71 VP1一致性达99%。EV71 VP1蛋白相对分子质量约为32×103,主要以包涵体形式存在。生物信息学分析得出,EV71 VP1蛋白为亲水性蛋白,无跨膜区,不具有N-端信号肽序列,存在三级结构。ELISA结果显示EV71阳性患者OD值为(2.425±0.521),COX A16阳性患者OD值为(1.205±0.314),健康对照组OD值为(0.353±0.128)。EV71 VP1蛋白检测敏感性和特异性分别为84%和88%。结论 成功构建了pET28a-EV71 VP1表达载体;通过对手足口患者血清进行ELISA初步分析,得出目的蛋白有较高的敏感性和特异性,初步证实具有生物学活性,可进一步用于EV71诊断及疫苗的相关研究。     

127例乙肝表面抗体ELISA检测与化学发光法的比较

Objective To compare the difference between ELISA and chemiluminescence assay in de-tecting hepatitis B surface antibody (HBsAb) serum valued in 5～100mIU/ml. Method 127 HBsAb serum samples that had valued by chemiluminescence were detected using 10 domestic ELISA kits. The correlation between the results of ELISA kits and chemiluminescence assay were analyzed. Coincidence between the results of ELISA kits and chemiluminescence at different OD value range were also analyzed. Results The detected results between 10 domestic ELISA kits and chemiluminescence assay are positive correlation (P<0.01); The optimal coincidence rates of the results of ELISA with those samples valued by chemiluminescence at more than 100mIU/ml was 99.4%; the coinci-dence rate is from 42% to 80% in cut-off±20%. The average coincidence rate is under 60%. Conclusions Though the detected results of 10 domestic ELISA reagents is related closely, the coincidence rate of the results at 5～10mIU/ ml in detecting anti-HBs is low. So it is important to make a positive standard in detecting anti-HBs.     

127例乙肝表面抗体ELISA检测与化学发光法的比较

Objective To compare the difference between ELISA and chemiluminescence assay in de-tecting hepatitis B surface antibody (HBsAb) serum valued in 5～100mIU/ml. Method 127 HBsAb serum samples that had valued by chemiluminescence were detected using 10 domestic ELISA kits. The correlation between the results of ELISA kits and chemiluminescence assay were analyzed. Coincidence between the results of ELISA kits and chemiluminescence at different OD value range were also analyzed. Results The detected results between 10 domestic ELISA kits and chemiluminescence assay are positive correlation (P<0.01); The optimal coincidence rates of the results of ELISA with those samples valued by chemiluminescence at more than 100mIU/ml was 99.4%; the coinci-dence rate is from 42% to 80% in cut-off±20%. The average coincidence rate is under 60%. Conclusions Though the detected results of 10 domestic ELISA reagents is related closely, the coincidence rate of the results at 5～10mIU/ ml in detecting anti-HBs is low. So it is important to make a positive standard in detecting anti-HBs.     

抗中心体抗体在儿童病原感染及自身免疫性疾病中的临床意义

目的初步了解抗中心体抗体(ACA)在儿童病原感染及自身免疫性疾病中的临床意义。方法收集广州市儿童医院2009年4月至9月19例ACA检测阳性患儿为ACA阳性组、42例ACA阴性为阴性对照组,选取同期常见病原微生物感染及正常儿童为阳性对照组和正常对照组。对4组病原微生物感染指标和其他自身抗体及炎症相关蛋白资料进行统计学分析。结果 ACA阳性存在于自身免疫相关疾病如系统性红斑狼疮(SLE)等16例次(84.2%);感染因素相关疾病7例次(36.8%)。ACA阳性组与阴性对照组间病原微生物感染指标差异无统计学意义(P>0.05);两组间ANA阳性率差异有统计学意义(P<0.001)。阳性对照组、正常对照组与ACA阳性组间病原微生物感染指标差异无统计学意义(P>0.05)。结论儿童ACA常见于免疫性疾病如SLE、川崎病,与机体亢进的自身免疫状态有关,而与以往报道的MP、CP、HSV等病原微生物感染无直接关系。     

127例乙肝表面抗体ELISA检测与化学发光法的比较

Objective To compare the difference between ELISA and chemiluminescence assay in de-tecting hepatitis B surface antibody (HBsAb) serum valued in 5～100mIU/ml. Method 127 HBsAb serum samples that had valued by chemiluminescence were detected using 10 domestic ELISA kits. The correlation between the results of ELISA kits and chemiluminescence assay were analyzed. Coincidence between the results of ELISA kits and chemiluminescence at different OD value range were also analyzed. Results The detected results between 10 domestic ELISA kits and chemiluminescence assay are positive correlation (P<0.01); The optimal coincidence rates of the results of ELISA with those samples valued by chemiluminescence at more than 100mIU/ml was 99.4%; the coinci-dence rate is from 42% to 80% in cut-off±20%. The average coincidence rate is under 60%. Conclusions Though the detected results of 10 domestic ELISA reagents is related closely, the coincidence rate of the results at 5～10mIU/ ml in detecting anti-HBs is low. So it is important to make a positive standard in detecting anti-HBs.     

722例婴幼儿病毒性肺炎的血清学研究

婴幼儿病毒性肺炎是儿童的严重呼吸道疾病之一。由于病毒性肺炎临床表现不突出,加之感染的病毒种类较多,这就给实验室提出了明确诊断的要求。我们曾在病原学方面作过多次报道。本文就1978～1984年的722例婴幼儿病毒性肺炎进行血清学研究,并与病毒分离进行比较,现报告如下。     

127例乙肝表面抗体ELISA检测与化学发光法的比较

目的 探讨浓度处于5～100mIU/ml的乙肝表面抗体(HBsAb)ELISA定性检测与化学发光法的符合情况.方法 采用10种国产主流HBsAb ELISA试剂对127例经用化学发光法检测已知HBsAb浓度的标本进行检测,分析ELISA检测OD值与抗体浓度的相关性,并以试剂盒临界值为准分别比较不同OD值段ELISA结果 与化学发光法结果 的符合率.结果 10种同产HBsAb ELISA试剂盒OD值与化学发光定最值均呈正相关(P<0.01);ELISA法检测结果 与化学发光的阴阳符合率比较显示,5～10mIU/ml样本符合率较低,只有37.5%,在大于100mIU/ml时可达99.4%,在10～30mIU/ml,30～100mIU/ml分别为75.3%和90.7%;OD值存0～0.05,0.05～0.105、0.105～0.21、0.21～0.3、0.3～0.5、大于0.5分别为73.4%、65.3%、60.5%、72.0%、86.5%、92.2%.结果 显示阴阳结果 符合率在较高在大于0.5时最高,在介于0.105～0.21的符合率最低;处于试剂盒临界值±20%处的标本的符合率普遍偏低,最低只有42%,最高亦不超过80%.结论 虽然10种国产ELISA试剂之间的检测结果 没有统计学差异,但是对于浓度介于5～10mIU/ml标本符合率低.而且在接近试剂盒临界值时与化学发光结果 符合率则很低,但是随着OD值的增高符合率也逐渐升高.考虑到乙肝表面抗体的特殊临床意义,如何选取一个更为科学客观的结果 判断临界值值得进一步探讨.     

反复呼吸道感染儿童血清中白介素2,4,8及受体2的检测

目的　观察反复呼吸道感染 (RRI)患儿血中部分细胞因子的状况 ,了解患儿机体的免疫功能状态。方法　采用ELISA双抗体夹心法对 70例RRI患儿及 2 0例正常儿童血清中的IL2、IL4、IL8、IL2R进行检测。结果　RRI患儿血清中IL2R比正常组明显上升 (P <0 0 1) ,IL2比正常组明显下降 (P <0 0 1) ,IL4比正常组明显上升 (P<0 0 5 )。IL2与IL2R成直线相关 (r=0 2 92 ,P <0 0 5 )。结论　RRI患儿机体体液免疫功能紊乱 ,TH1/TH2细胞因子偏移与儿童RRI的发病有关。     

广州地区1～8岁健康儿童补体C3,C4含量的检测及其参考范围的建立

目的建立广州地区1～8岁儿童血清补体C3,C4含量的参考范围。方法应用Beckman—coulter Immage800特种蛋白分析仪,采用免疫比浊法对968例1～8岁健康儿童血清补体C3,C4含量进行检测,分成7个年龄段对结果进行统计分析。结果广州地区1～8岁健康儿童血清补体C3,C4含量在性别和各年龄段之间差别无统计学意义（P〉0．05）,总的参考范围（g／L）为C3：0．59～1．81,C4：0．19～0．22。同时与全国临床检验操作规程以及厂家提供的参考范围相比较差异均有统计学意义（P〈0．05）。结论1～8岁儿童血清补体C3,C4旧的参考范围已不适用于当今人群,各地、各实验室应测定一批健康人的标本,统计出各自实验室的参考范围,若存在性别差异,应分别制定两性的参考范围,若存在年龄组间差异,还应进一步制定两性间各个年龄组的参考范围。