流式微球载体技术在检测肾综合征出血热患者血清特异性抗体和细胞因子中的应用

目的:建立流式微球载体技术(FMA)检测肾综合征出血热(HFRS)患者血清抗HFRS 病毒特异性抗体IgM和IgG及细胞因子含量的新方法.方法:选择28例临床确诊的HFRS患者及20例健康人血清标本,FMA定量检测抗HFRS 病毒IgM和IgG;定量检测细胞因子IL-6和TNF-α.检测结果与ELISA法进行比较.结果:FMA检测HFRS患者抗HFRS病毒IgM和IgG的阳性率分别为92.85%和71.43%,健康对照组的抗体阳性率(假阳性率)为0;HFRS患者血清IL-6和TNF-α的含量分别为(532.62±397.19) ng/L和(392.68±177.68) ng/L,明显高于健康对照组(38.77±20.32)ng/L (P<0.01)和(15.91±6.91) ng/L(P<0.01).ELISA法检测HFRS患者抗HFRS病毒IgM和IgG的阳性率分别为71.43% 和50.00%,健康对照组的抗体阳性率(假阳性率)为0;HFRS患者血清IL-6和TNF-α的含量分别为(256.46±102.51) ng/L和(45.63±5.32) ng/L,高于健康对照组(53.8±19.21) ng/L(P<0.01)和(5.81±3.58) ng/L(P<0.01).结论:建立了FMA法对HFRS患者的特异性抗体和IL-6和TNF-α的检测,其灵敏度明显优于ELISA法,为HFRS的临床诊断和病理机制研究提供了新的方法.     

高血压患者人巨细胞病毒感染率及其血浆中和抗体水平分析

目的 对高血压患者人巨细胞病毒（HCMV）感染率及其血浆中和抗体水平展开研究分析,研究高血压与人巨细胞病毒感染的相关性.方法 随机选取2011年12月-2013年12月期间接收治疗的50例高血压患者血标本作为观察组,同时选取50例健康体检人员的血标本作为对照组.对两组标本人巨细胞病毒特异性中和抗体、人巨细胞病毒特异性IgG和IgM、人巨细胞病毒特异性UL93 DNA进行检测.结果 观察组HCMV UL93 DNA的阳性率为72.0％,HCMV IgG阳性率为70.0％,HCMV IgM阳性率为4.0％;对照组HCMV UL93 DNA阳性率为54.0％,HCMV IgG阳性率为52.0％,HCMV IgM阳性率为2.0％;观察组HCMV UL93 DNA阳性率、HCMV IgG阳性率显著高于对照组,数据差异具有统计学意义（P＜0.05）;两组HCMV IgM阳性率数据对比差异无统计学意义（P＞0.05）.结论 高血压患者人巨细胞病毒感染率显著高于健康对照组,但特异性中和抗体水平较健康对照组显著下降,即高血压患者的人巨细胞病毒感染体液免疫状态不足,高血压与人巨细胞病毒感染显著相关.     

妊高征患者血浆P-选择素水平的变化

目的探讨妊高征患者血浆P-选择素含量的变化及其在妊高征发病中的意义。方法采用酶联免疫吸附法测定50例妊高征患者（HDP组）,其中妊娠期高血压组12例,轻度子痫前期组20例,重度子痫前期组18例、50例正常晚孕妇女（对照组）血浆P-选择素。结果 HDP组血浆P-选择素值（75.26±24.56μg/l）显著高于对照组（46.24±12.02μg/l）（P〈0.001）,以子痫前期组增高明显。结论妊高征患者血浆P-选择素水平含量变化与妊高征发病及病情发展有关。     

内皮细胞分化功能在替米沙坦干预高血压患者过程中的变化

背景:研究表明,原发性高血压患者内皮细胞受损的特征性改变是内皮素释放失衡。替米沙坦能够恢复高血压患者的"杓型"血压模式,其降压效果优于氯沙坦、氨氯地平等常规药物,有更强的保护作用。目的:分析替米沙坦对高血压患者内皮细胞分化功能及血清脂联素的影响。方法:160例原发性高血压患者按抽签法分为治疗组与对照组各80例,对照组给予海捷亚治疗,治疗组给予替米沙坦治疗,1片/d,晨口服,治疗周期为4周。观察两组患者临床疗效、血浆脂联素、血浆内皮素及一氧化氮值变化,并进行血浆脂联素与高血压患者的血浆内皮素与一氧化氮值的相关性分析。结果与结论:治疗组的有效率为92.5%,对照组为75.0%,组间对比差异明显(P0.05)。两组患者治疗后的血浆脂联素和一氧化水平量都明显上升,血浆内皮素明显降低,与治疗前对比差异有显著性意义(P0.05);治疗后治疗组内皮素显著低于对照组,一氧化氮显著高于对照组(均P0.05)。Pearson相关检验分析显示血浆脂联素与血浆内皮素值呈现显著负相关(P0.05),与血浆一氧化氮值呈现显著正相关(P0.05)。说明替米沙坦治疗高血压能提高预后疗效,其作用的发挥与改善内皮细胞分化与提高血清脂联素有一定的相关性。     

阻塞性睡眠呼吸暂停低通气综合征患者细胞粘附分子水平的研究

目的探讨细胞粘附分子在阻塞性睡眠呼吸暂停低通气综合症(OS-AHS)发病中的作用。方法应用酶联免疫吸附法(ELISA)检测30例老年OSAHS患者及30例老年健康对照者血清可溶性细胞间粘附分子-1(ICAM-1)、血管细胞粘附分子-1(VCAM-1)和E-选择素的含量。结果OSAHS组血清ICAM-1、VCAM-1、E-选择素含量分别为245.22±71.19ng/ml、24.01±4.79ng/ml、和86.58±48.02ng/ml,均明显高于健康对照组(P<0.01),且随OSAHS程度的加重而明显升高。ICAM-1、E-选择素水平与睡眠呼吸暂停低通气指数(AHI)呈明显正相关(P<0.01);I-CAM-1水平与最低血氧饱和度(SaO2min)呈明显负相关(P<0.01)。结论OSAHS患者血清中可溶性粘附分子ICAM-1、VCAM-1和E-选择素水平可作为反映OSAHS严重程度的敏感指标。     

巨细胞病毒感染与急性冠状动脉综合征患者血清sP-选择素和肿瘤坏死因子的变化

目的　探讨人巨细胞病毒(HCMV)感染与急性冠状动脉综合征(ACS)患者血清sP 选择素、肿瘤坏死因子(TNF α)的变化及相关性。方法　采用酶联免疫吸附技术检测79例ACS患者、30例稳定性心绞痛(SA)患者和30例正常对照组血清HCMV IgM、HCMV IgG、sP 选择素和TNF α的水平。结果　(1)在ACS、SA及正常对照组中,血清HCMV IgM、IgG的阳性率分别为30 4 % (2 4 79)、10 0 %(3 30 )和6 7% (2 30 ) ;86 1% (6 8 79)、80 0 % (2 4 30 )和5 3 3% (16 30 )。HCMV IgM的阳性率在ACS组中明显高于SA组和对照组(P <0 0 1) ,HCMV IgG的阳性率在ACS和SA组中明显高于对照组(P <0 0 1)。(2 )与SA组和对照组相比,ACS组中血清sP 选择素和TNF α水平显著升高,分别为(15 2 0 0±112 7)和(14 81 0±10 9 1)pg ml比(6 4 37 3±6 6 6 9)pg ml,(2 7 3±13 7)pg ml和(2 8 1±11 3)pg ml比(5 6 2±18 4 )pg ml,组间差异有统计学意义(P <0 0 1)。随着冠状动脉(冠脉)病变程度的增加,ACS中急性心肌梗塞(AMI)组与不稳定性心绞痛(UA)组相比,sP 选择素和TNF α水平显著升高(P <0 0 1) ,而SA组与对照组间差异无统计学意义(P >0 0 5 )。(3)ACS组中HCMV IgM阳性患者血清中sP 选择素和TNF α水平较HCMV IgM阴性患者明显升高(P <0 0     

原发性高血压患者治疗前后血浆ET和E选择素检测的临床意义

目的：探讨了原发性高血压患者治疗前后血浆ET和E选择素水平及临床意义。方法：应用放免法检测了39例原发性高血压患者治疗前后血浆ET水平,酶联免疫法检测E选择素水平,并与35名正常健康人作比较。结果：原发性高血压患者治疗前后血浆ET和E选择素水平非常显著地高于正常人组水平（P〈0．01）,经治疗3个月后则与正常人组比较无显著性差异（P〉0．05）。结论;血浆ET和E选择素的异常升高是原发性高血压发病的病理因素之一,对诊断、治疗和预后均有重要的临床价值。     

细胞因子及特异性抗体的检测在肾综合征出血热诊断中的意义

探讨检测血清细胞因子及肾综合征出血热(HFRS) 病毒特异性抗体IgM和IgG的含量在HFRS发病机制及诊断中的意义.选择24例HFRS患者及30例健康人血清标本,采用生物素-亲和素-酶免疫技术检测IL-2、IL-6和TNF-α,ELISA方法检测血清HFRS病毒特异性抗体IgM和IgG,并对其进行统计学分析. 结果显示, ELISA法检测HFRS患者抗HFRS病毒IgM和IgG的阳性率分别为75.00 % 和50.00 %,健康对照组的抗体阳性率为零;HFRS患者血清IL-2、IL-6、TNF-α的含量分别为10.88±2.31pg/mL、256.46±102.51pg/mL和45.63±5.32pg/mL,高于健康对照组0.59±0.24pg/mL(P＜0.01)、53.8±19.21 pg/mL(P＜0.01)和5.81±3.58 pg/mL(P＜0.01). 结论 :HFRS患者血清IL-2、IL-6和TNF-α及血清特异性抗体IgM和IgG的含量较健康人明显升高,检测这些指标对该病发病机理、诊断及预后评价有一定意义.     

1测定醛固酮浓度/肾素活性(SAC/PRA)值在原发性醛固酮增多症中的临床意义

目的:探讨测定SAC/PRA值对原发性醛固酮增多症的诊断价值.方法:采用放射免疫分析法测定48例原发性醛固酮增多症患者和30例正常人的血浆肾素(PRA),血管紧张素Ⅱ(AT-Ⅱ)以及血清醛固酮(Aldo),并计算醛固酮浓度/肾素活性(SAC/PRA)比值.结果:正常组PRA、AT-Ⅱ、Aldo测定值分别为0.57±0.08ng/ml/h,36.03±6.11ng/L,0.33±0.04nmol/L;原醛患者PRA、AT-Ⅱ、Aldo测定值分别为0.14±0.08ng/ml/h,21.21±7.55ng/L,1.07±0.34nmol/L.与正常对照组比较,均有极显著性差异(p＜0.001).SAC/PRA(ng/dl/ng*ml-1*h-1)913±409.结论:合理使用SAC/PRA比值有助于原发性醛固酮增多症的诊断.     

抗IgG、IgM、IgA型心磷脂抗体和抗β_2糖蛋白1抗体在系统性红斑狼疮患者中的意义

目的探讨抗IgG、IgM、IgA型心磷脂抗体和抗β2糖蛋白1抗体在系统性红斑狼疮(systemic lupus erythematosus,SLE)患者中的意义。方法分别采用化学发光免疫分析法和酶联免疫吸附法检测372例SLE患者、80例其他结缔组织病患者及60例健康体检者血清中抗IgG、IgM、IgA型心磷脂抗体和抗β2糖蛋白1抗体。同时分析这4个指标与SLE及其病情严重程度的相关性。结果在检测的372例SLE患者组中,抗IgG、IgM、IgA型心磷脂抗体和抗β2糖蛋白1抗体的阳性率分别为31.45%、14.52%、10.22%和29.03%,其表达水平分别为(90.39±35.43)U/ml、(41.25±23.16)U/ml、(32.27±15.77)U/ml、(61.42±21.69)U/ml,与SLEDAI积分呈正相关(P0.05)。在80例疾病对照组中,抗IgG、IgM、IgA型心磷脂抗体和抗β2糖蛋白1抗体的阳性率分别为7.50%、2.50%、0和1.25%。而在60例正常对照组中,4项检测结果均为阴性。抗IgG型心磷脂抗体阳性与血小板减少、白细胞减少、血栓形成、习惯性流产和肾脏病变有关;抗IgM型心磷脂抗体阳性与血小板减少、白细胞减少、血栓形成和肾脏病变有关;抗IgA型心磷脂抗体阳性与血小板减少、血栓形成和肾脏病变有关;抗β_2糖蛋白1抗体阳性与血小板减少、白细胞减少、溶血性贫血、血栓形成、习惯性流产和肾脏病变有关。结论抗IgG、IgM、IgA型心磷脂抗体和抗β_2糖蛋白1抗体在SLE的发病及病情发展中起到了非常重要的作用,定量和联合检测对SLE的诊断、病情严重程度的评价、判断预后及疗效观察等均有重要的临床意义。     

原发性高血压患者巨细胞病毒感染及与血管并发症关系 …

目的 探讨原发性高血压患者巨细胞病毒感染及与血管并发症之间的关系。方法 应用间接酶联免疫吸附试验（ＥＬＩＳＡ）检测了１０５例原发性高血压患者血清人巨细胞病毒（ＨＣＭＶ）特异性抗体。结论 研究结果提示原发性高血压患者存在较高的活动性巨细胞病毒感染，ＨＣＭＶ活动性感染似与血管并发症有一定关联。     

巨细胞病毒感染与可溶性白细胞介素2受体的关系

应用酶联免疫吸附试验（ＥＬＩＳＡ）对１０４例育龄妇女的血清进行了巨细胞病毒（ＨＣＭＶ）ＩｇＧ、ＩｇＭ抗体的检测，同时用ＥＬＩＳＡ双抗体夹心法测定了不同感染状态下血清中可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）的水平，并将ｓＩＬ－２Ｒ水平与未感染ＨＣＭＶ的正常育龄妇女进行了比较。结果，育龄妇女中抗－ＨＣＭＶＩｇＧ的阳性率为８９．４％，ＩｇＭ的阳性率为９．６％，感染ＨＣＭＶ的妇女血清中ｓＩＬ－２Ｒ水平均大于未感染的对照组（１７８．１±５７．３Ｕ／ｍｌ），Ｐ＜０．０５，其中ＩｇＭ阳性者和ＩｇＭ、ＩｇＧ同时阳性者血清中ｓＩＬ－２Ｒ水平最高，分别为９１０±４６５．６Ｕ／ｍｌ和９０５±３４７．８Ｕ／ｍｌ，两者间的差异无显著性意义（Ｐ＞０．０５），但均大于仅抗－ＨＣＭＶＩｇＧ阳性者（４４６．８±１５８．９Ｕ／ｍｌ），Ｐ均＜０．０５。表明，ＨＣＭＶ感染可致ｓＩＬ－２Ｒ水平升高，并且活动性感染者上升明显。提示：ｓＩＬ－２Ｒ可能参与了ＨＣＭＶ的免疫致病机制。     

温州市区育龄妇女孕前巨细胞病毒感染现状调查

目的了解温州地区育龄妇女孕前人巨细胞病毒（HCMV）感染的状况。方法收集2008年10月至2010年6日参加温州市龙湾区免费孕前优生筛查的妇女血标本2869份,采用酶联免疫吸附试验（ELISA）检测血清HCMV IgG/IgM抗体;HCMV IgM抗体阳性标本,采用实时荧光定量聚合酶链反应（FQ-PCR）检测血HCMV DNA载量;HCMV IgG/IgM抗体双阳性标本,采用尿素变性结合ELISA技术检测IgG抗体亲和力指数（AI）。结果 2869份孕前妇女血清中HC-MV IgG抗体阳性检出率为97.77%（2805/2869）,HCMV IgM抗体阳性检出率为0.77%（22/2 869）,IgG/IgM抗体均阳性检出率占0.17%（5/2 869）;22份HCMV IgM阳性标本中,血HCMV DNA阳性检出率为68.18%（15/22）;5份HCMVIgG/IgM双阳性标本中,检出低亲和力IgG抗体1份,中等亲和力IgG抗体2份,高亲和力IgG抗体2份。结论温州市区育龄妇女孕前HCMV IgG抗体阳性率高;对HCMV IgM抗体阳性孕前妇女应进行多指标检测以判断HCMV感染的状态,为减少出生缺陷、做好优生优育服务提供依据。     

Screening of blood donors for human cytomegalovirus (HCMV) IgG antibody with an enzyme immunoassay using recombinant antigens.

BACKGROUND: Screening of blood donors for human cytomegalovirus (HCMV) infection is usually performed by the combined detection of specific IgG and IgM antibody. However, in most of the cases of primary infection HCMV IgG seroconversion is observed concomitantly to IgM production and HCMV IgM antibody detection for blood donor screening is subject to a relatively high frequency of false positive results. OBJECTIVE: In the present study a newly established HCMV IgG ELISA based on recombinant antigens (anti-HCMV recombinant IgG ELISA, Biotest) was evaluated in terms of sensitivity and specificity for blood donor screening. STUDY DESIGN: A total of 442 serum samples including follow-up sera of five patients suffering from primary HCMV infection, selected seropositive and seronegative blood donors and routine specimens were comparatively investigated with three HCMV antibody ELISAs (anti-HCMV recombinant IgG ELISA, Biotest; Enzygnost anti-CMV/IgG + IgM, Dade Behring; and Captia CMV-TA, Centocor). RESULTS: IgG seroconversion was detected with anti-HCMV recombinant IgG ELISA as early as IgM in all five patients suffering from primary infection. The alternative ELISAs were less sensitive, detecting seroconversion one to three bleeds later in 2 (Enzygnost anti-CMV/IgG + IgM) and 4 patients (Captia CMV-TA), respectively. Anti-HCMV recombinant IgG ELISA showed a 99.1% agreement with Enzygnost anti-CMV/IgG + IgM and/or Western blot in the preselected blood donors and routine specimens. Relatively high numbers of false negative (n=20) and positive results (n=7) were obtained with Captia CMV-TA. CONCLUSIONS: Our preliminary data suggest that HCMV antibody screening of blood donors can be performed reliably by detection of specific IgG provided that a highly sensitive assay system is used.     

Cytomegalovirus DNA detection in sera from patients with active cytomegalovirus infections.

Using a specific and sensitive polymerase chain reaction method, we detected reliably the presence of human cytomegalovirus (HCMV) DNA directly in serum samples collected at an early stage of HCMV infection, even before immunoglobulin M (IgM) antibodies were measurable. HCMV DNA was detected in serum from all patients with active HCMV infection; in 91% of these patients, HCMV DNA was found in the acute-phase serum. In 13 of 44 patients, HCMV DNA was found in serum before HCMV-specific IgM. For four kidney transplant recipients, the occurrence of HCMV DNA in serum, virus isolation from urine and leukocytes, and HCMV IgG and IgM serology were determined. We found a correlation between HCMV DNA in serum and positive virus isolation from leukocytes. In three of five congenitally infected infants, HCMV DNA and HCMV IgM were detected in the same sample. Two other infants were HCMV DNA positive, although no HCMV IgM antibodies were measurable. HCMV was found in urine from these infants either by virus isolation or with the polymerase chain reaction. Serum from one of the 22 healthy HCMV-seropositive blood donors was HCMV polymerase chain reaction positive.     

Evaluation of virological procedures to detect fetal human cytomegalovirus infection: Avidity of IgG antibodies,virus detection in amniotic fluid and maternal serum

Human cytomegalovirus (HCMV) is the most common cause of viral intrauterine infection and fetal damage largely due to maternal primary infection. Virological procedures which are able to detect HCMV fetal infection were evaluated. HCMV IgG antibodies were detected in 62.5% of the pregnant women and 1.47% had a primary infection. From March, 1992 to August, 1995, 29 seroconversions were observed, and in 64 other cases. HCMV IgM antibodies were detected in the first serological test. The mean IgG antibody avidity test (AI) was 31% for the 11 seroconversions tested and 74% in 32 cases where IgG and IgM HCMV antibodies were detected in the first serum. In the 29 HCMV seroconversions, 19 amniocentesis were carried out and 12 fetuses (41.4%) were infected in utero. In four amniotic fluids positive in culture and PCR, the fetus or newborns were infected and in one out of the two cordocentesis undertaken, hepatitis, anemia, and thrombocytopenia were noted. In four other cases, investigations seeking HCMV in amniotic fluid were negative whereas infants were infected at birth. Among the 64 cases with positive HCMV IgM and IgG antibodies detected in the first serological test, three fetuses were infected in utero, but no amniotic fluid was available in these cases. Amniotic fluids were studied in 39 cases, and HCMV detection by culture and PCR-hybridization was negative. HCMV DNA was detected in the maternal sera of five out of 21 pairs of seroconversions and in two cases on the first negative serum. The assay was also carried out on 50 of the 64 HCMV IgM positive sera. Two had detectable HCMV DNA. © 1996 Wiley-Liss, Inc.     

1995至2000年北京地区散发性急性肝炎患者肝炎病毒抗体的检测与分析

目的 了解北京市地区散发性肝炎患者甲型、乙型、丙型和戊型肝炎病毒感染型别分布及重叠感染。方法 用EIA法检测1995斫2月至2000年12月北京地区散发性急性肝炎患者抗－HAVIgM、HBsAg／抗－HBcIgM、抗－HCVIgM/IgG和抗－HEVIgM/IgG。结果 214例散发性急性肝炎患者血清,抗甲、乙、内和戊型肝炎病毒IgM总阳性数155例,戊型肝炎76例。有9名患者检出2种肝炎病毒抗原或抗体阳性,其中在3名肝硬化患者和2名静脉吸毒者同时检测到HBV和HCV抗原或抗体,1例HBsAg阳性者检测到抗－HAVIgM,3例－HCVIgG阳性者中分别检测到2例抗－HBVIgM和1例抗－HEVIgM。肝炎病毒重叠感染的9名患者年龄在31岁至49岁之间。结论 北京地区散发性急性肝炎78％是由消化道传染的甲戊型肝炎病毒引起,戊型肝炎在四种肝炎中位居首位,其次为甲型肝炎、乙型肝炎和丙型肝炎。肝炎病毒重叠感染多见乙、丙型肝炎病毒合并感染或慢性乙、丙型肝炎患者合并甲型或戊型肝炎病毒感染。在我国对散发性病毒性肝炎的预防应引起高度重视。     

Anti-human cytomegalovirus nuclear antigen antibodies of different immunoglobulin classes

IgG, IgM and IgA immunoglobulin classes of antibodies to human cytomegalovirus nuclear antigens (CMNA) were studied by the acid-fixed nuclear binding technique (AFNB) and combined anti-complement immunofluorescence (combined ACIF). In acute cases of infectious mononucleosis (IM) of human cytomegalovirus (HCMV) origin and in the so-called double virus infections (HCMV + Epstein-Barr virus), anti-CMNA IgM antibodies were detected. They were absent from both anti-HCMV positive sera of healthy donors and sera of patients suffering of IM caused by EBV used as controls. The presence of anti-CMNA IgM may thus serve as an additional evidence of acute HCMV infection. Non-complement-fixing IgA classes of the anti-CMNA antibodies were not found in some of the sera gathered during the acute phase of IM of EBV origin: in one fourth of the HCMV seropositive donors and in a number of late serum samples. But non-complement-fixing and complement-fixing anti-CMNA components of the IgG class were detected.