Monitoring of human cytomegalovirus glycoprotein B genotypes using real-time quantitative PCR in immunocompromised Chinese patients

Based on sequence variation in the N-terminus of glycoprotein B (gB), human cytomegalovirus (HCMV) can be classified into four gBn genotypes, and these genotypes are associated with different clinical outcomes. The distribution of gBn genotypes and the level of gBn DNA load were examined in immunocompromised Chinese patients using real-time quantitative PCR. In addition, the PCR and pp65 antigenemia results were compared. In 1480 specimens, 81.4% were antigen-positive, 12.6% were PCR-positive. The gB genotype distribution was as follows among PCR-positive samples: gBn1, 63.1%; gBn2, 13.4%; gBn3, 8.6%; gBn4, not detected; mixed genotypes, 14.9% (gBn1 and gBn3, 14.4%; gBn2 and gBn3, 0.5%). The gBn3 and gBn1 genotypes had the highest and lowest copy numbers, respectively (p < 0.05). The quantity of gBn DNA found in PCR-positive, pp65-negative samples was significantly lower than that found in PCR-positive, pp65-positive samples (p < 0.05). The PCR and antigenemia results did not differ among bone marrow transplant patients, solid organ transplant patients, and immunocompromised patients without transplantation (p > 0.05). HCMV gBn genotyping using real-time quantitative PCR was established successfully, and the distribution of gBn genotypes in immunocompromised Chinese patients was investigated. This method may help to understand better the relationship between gBn genotype and clinical outcome and aid in clinical detection.     

Development of a novel,guinea pig-specific IFN-γ ELISPOT assay and characterization of guinea pig cytomegalovirus GP83-specific cellular immune responses following immunization with a modified vaccinia virus Ankara (MVA)-vectored GP83 vaccine

The guinea pig (Cavia porcellus) provides a useful animal model for studying the pathogenesis of many infectious diseases, and for preclinical evaluation of vaccines. However, guinea pig models are limited by the lack of immunological reagents required for characterization and quantification of antigen-specific T cell responses. To address this deficiency, an enzyme-linked immunospot (ELISPOT) assay for guinea pig interferon (IFN)-γ was developed to measure antigen/epitope-specific T cell responses to guinea pig cytomegalovirus (GPCMV) vaccines. Using splenocytes harvested from animals vaccinated with a modified vaccinia virus Ankara (MVA) vector encoding the GPCMV GP83 (homolog of human CMV pp65 [gpUL83]) protein, we were able to enumerate and map antigen-specific responses, both in vaccinated as well as GPCMV-infected animals, using a panel of GP83-specific peptides. Several potential immunodominant GP83-specific peptides were identified, including one epitope, LGIVHFFDN, that was noted in all guinea pigs that had a detectable CD8+ response to GP83. Development of a guinea pig IFN-γ ELISPOT should be useful in characterization of additional T cell-specific responses to GPCMV, as well as other pathogens. This information in turn can help focus future experimental evaluation of immunization strategies, both for GPCMV as well as for other vaccine-preventable illnesses studied in the guinea pig model.     

新生儿G-6-PD缺陷症合并巨细胞病毒感染的研究

了解新生儿G-6-PD缺陷症合并巨细胞病毒感染的情况.方法对新生儿高胆红素血症患儿进行G-6-PD活性、PCR HCMV及其他相关检查.结果新生儿G-6-PD缺陷症病人合并HCMV(+)者其ALT、IBIL、DBIL、血红蛋白、胆汁酸、G-6-PD活性与新生儿G-6-PD缺陷症HCMV(-)者、非G-6-PD缺陷症HCMV(+)者、非G-6-PD缺陷症HCMV(-)者比较差异均有显著性.结论新生儿G-6-PD缺陷症合并巨细胞病毒感染会加重肝脏、血液系统的损害,巨细胞病毒感染可降低新生儿G-6-PD活性.     

性病和不育不孕患者巨细胞病毒感染状况调查分析

目的　调查性病和不育不孕症患者巨细胞病毒感染状况 ,探讨性病—HCMV感染—不育不孕症的关系。　方法　采用酶联免疫吸附法 (ELISA)检测性病、不育不孕症患者和健康对照组血清中HCMV—IgM、IgG特异抗体 ,测定其活动性感染、既往性感染和总感染率。　结果　性病组与不育不孕组的HCMV总感染率分别为 94 73 3 %、90 2 2 % ,与对照组的 5 8 5 0 %相比较P <0 0 1。各组性病的HCMV总感染率分别为 97.14 %、94.67%、94.76%、94.64 %、92 .86% ,分别与对照组相比较差异具显著性P <0 .0 1。不育不孕组中有性病与无性病两组的HCMV总感染率分别为94 73 %和 83 .15 % ,两组之间相比较差异有显著性P <0 0 1,分别与对照组相比较P <0 0 1。　结论　性病—HCMV感染—不育不孕症三者之间存在密切联系 ,HCMV通过性传播途径感染可导致不育不孕症 ,必须加强性健康教育并对未生育和性病患者及性病感染高危人群进行CMV感染的检测 ,提高生育质量     

对献血者巨细胞病毒感染的研究

目的 了解献血中巨细胞病毒(HCMV)感染状况，并探索一种比较简便的检测方法。方法 用PCR法和DNA杂交法检测同一献血的白细胞及血清中的HCMV—DNA，并用ELISA法检测血清中的HCMV-IgM、IgG(测4个滴度)，连续2a共检测白细胞和血清样本各200人份。结果 PCR法检测白细胞中的HCMV-DNA阳性率分别为63％和70％，DNA杂交法检测的阳性率分别为42％和50％；PCR法检测血清中的HCMV-DNA的阳性率分别为49％和53％，DNA杂交法检测的阳性率分别为33％和39％；HCMV-IgM阳性率2a均为5％；HCMV-IgG阳性率分别为54％和58％。结论 HCMV-IgG抗体可作为HCMV感染的指标。     

Lack of association between the kinetics of human cytomegalovirus (HCMV) glycoprotein B (gB)-specific and neutralizing serum antibodies and development or recovery from HCMV active infection in patients undergoing allogeneic stem cell transplant

The kinetics of the gB-specific and neutralizing antibody responses to human cytomegalovirus (HCMV) were analyzed in 26 allogeneic stem-cell transplant recipients who either did (n = 20) or did not (n = 6) develop asymptomatic HCMV active infection during the study period. Antibody response profiles varied widely among individuals in both groups, irrespective of whether HCMV active infection did or did not occur. Development of HCMV active infection was not preceded by a decline in functional serum antibody levels. Neither the absence nor the presence of HCMV active infection correlated with either high or low serum levels of gB-specific and neutralizing antibodies, respectively. In most patients, episodes of HCMV replication were not followed by a marked increment in functional serum antibody titers. Therefore, resolution of an ongoing HCMV active infection was not associated with a vigorous antibody response to viral replication. In addition, this study supports previous data indicating that passive transfer of human immunoglobulins may result in an increment in gB-specific and neutralizing serum antibody levels, the magnitude of which varies among recipients; however, both patients with and without measurable increments in serum antibody levels developed HCMV active infections with comparable frequency.     

更昔洛韦联合胸腺肽治疗小儿巨细胞病毒感染疗效分析

目的探究更昔洛韦联合胸腺肽在HCMV感染的小儿患者治疗中的临床应用价值。方法采用前瞻性开放研究,共收集80例在我院就诊的人类巨细胞病毒(HCMV)小儿患者,分别进行更昔洛韦联合胸腺肽治疗和单用更昔洛韦治疗,经统计学分析比较两组治疗的总有效率和不良反应发生率。结果将二者总有效率和不良反应率采用SPSS18.0统计软件进行卡方检验后,差异均有统计学意义(P值均<0.05)。结论更昔洛韦与胸腺肽联合治疗小儿巨细胞病毒感染的疗效明显优于单用更昔洛韦治疗,且其不良反应明显低于后者,因此值得在临床上推广应用。     

巨细胞病毒感染婴儿母亲携带病毒检测的意义

目的探讨巨细胞病毒感染婴儿中人巨细胞病毒母乳感染率及意义。方法应用荧光定量PCR法检测124例确诊巨细胞病毒感染患儿母亲的母乳和全血中HCMV-DNA含量,并同时测定母亲血浆中HCMV-IgM。结果 FQ-PCR检测HCMVDNA乳汁的阳性率为69.4%,全血的阳性率为23.4%;HCMV-IgM阳性率27.4%。母乳HCMV-DNA的阳性率明显高于全血HCMV-DNA及特异性抗体IgM的阳性率(P<0.05)。结论感染巨细胞病毒患儿的母乳中带毒率高,母乳感染HCMV可能是婴儿获得性巨细胞感染的主要途径。     

HCMVpp65抗原、HCMV mRNA和IgM抗体检测在诊断HCMV活动性感染中的比较

目的比较人巨细胞病毒（HCMV）pp65抗原、HCMV mRNA和血清HCMV—IgM抗体检测3种方法在诊断HCMV活动性感染中的实用意义。方法采集医院TORCH检查HCMV—IgM阳性的病人外周血（60份）。将标本分2份2ml和3ml,别用于HCMV mRNA和pp65检测。将三者结果进行比较。结果pp65抗原检测的结果与IgM抗体检测的阳性符合率为81．67％。与HCMV mRNA检测相比pp65抗原检测法的符合率、特异度和敏感度分别为81．67％,81．81％和81．63％。而且高pp65抗原血症与患者的临床症状密切相关。结论pp65抗原血症反映该病毒活动状况,可监测HCMV活动性感染,联合HCMV—IgM的检测可以提高临床的诊断率并可用于指导临床用药及监测药物疗效。     

应用ELISA法检测119对母婴血中巨细胞病毒

人类巨细胞病毒（ＨＣＭＶ）是引起人类先天性畸形的重要原因之一。本文应用ＥＬＩＳＡ法对鞍山市的１１９对母婴止进行了ＨＣＭＶ－ＩｇＭ、ＨＣＭＶ－ＩｇＡ的检测。检测表明，鞍山市孕母ＨＣＭＶ近期复发及原发感染率为２９．４％，新生儿ＨＣＭＶ先天感染率为１３．４％，具有一定的感染性。同时检测ＨＣＭＶ－ＩｇＭ、ＩｇＡ具有很好的互补性，可以提高感染的检出率。