FQ—PCR血、尿、乳汁联合检测在小儿人巨细胞病毒感染中的临床研究

目的探讨用荧光定量聚合酶链反应（fluorescence quantitative PCR,FQ—PCR）法进行血、尿、母乳联合检测小儿人巨细胞病毒（HCMV）的临床意义。方法采用FQ—PCR法联合检测182例疑诊HCMV感染的患儿,对母乳喂养的阳性患儿补测母乳HCMV—DNA。结果血、尿及联合检测HCMV—DNA阳性率分别为15．93％、29．12％、39．01％。阳性患儿配对的母乳HCMV—DNA阳性率为45．00％,阴性患儿配对的母乳HCMV—DNA阳性率为18．60％,两组进行x^2检验,x^2=5．54,具有显著性差异（P〈0．02）。结论联合检测能提高HCMV—DNA检测阳性率,尿HCMV—DNA检测阳性率高于血HCMV—DNA检测,HCMV感染母乳是婴儿获得性感染的主要途径之一。     

新生儿黄疸中不同标本人巨细胞病毒检测及临床特征

目的:分析新生儿黄疸中不同类型标本人巨细胞病毒(HCMV)的检测结果;研究HCMV感染性新生儿黄疸的临床治疗情况。方法:选取2018年2月至2019年2月庆阳市人民医院住院的病理性黄疸新生儿300例,用实时荧光定量PCR(RT-FQ-PCR)方法检测患儿血浆、外周血单个核细胞(PBMC)、尿液HCMV-DNA及母乳HCMV-DNA,ELISA方法检测患儿血清HCMV-IgM和IgG,并用熊去氧胆酸片联合蓝光照射对症治疗,同时收集病理性黄疸患儿的临床数据和肝功能指标,对所有数据进行统计学分析。结果:300例黄疸患儿中,感染HCMV的为40例(13.33%),其中血浆HCMV-DNA阳性12例(4.00%),PBMC HCMV-DNA阳性20例(6.67%),尿液HCMV-DNA阳性30例(10.00%),母乳HCMV-DNA阳性138例(46.00%),血清HCMV-IgM阳性13例(4.33%),HCMV-IgG阳性228例(80.85%)。新生儿黄疸组血浆、PBMC、尿液、母乳HCMV-DNA及血清HCMV-IgM的阳性率显著高于正常对照组(P0.05)。HCMV感染组治疗后TBil、DBil、TBA、γ-GGT、AST、CK、CK-MB均有不同程度的降低,与对照组比较差异均有统计学意义(P0.05)。结论:新生儿病理性黄疸与HCMV感染密切相关;母乳HCMV-DNA阳性率最高,其次为尿液;熊去氧胆酸片联合蓝光照射对HCMV感染性新生儿黄疸有良好的治疗效果。     

尿液HCMV-DNA联合血清HCMV-IgM检测在患儿HCMV感染中的诊断价值分析

目的:分析尿液HCMV-DNA联合血清HCMV-IgM检测在患儿HCMV感染中的诊断效能.方法:选取本院2019年 7月-2022年 8 月收治HCMV感染患儿112 例.据主诉、体格检查及第一临床诊断将HCMV感染患儿分为肺炎组(n=68)、肝炎组(n=26)及其他组(n=18,主诉主要为发热、腹泻等).对比分析不同性别、疾病组、年龄尿液HCMV-DNA、血清HCMV-IgM检测结果.结果:尿液HCMV-DNA阳性率为30.35%、血清HCMV-IgM阳性率为33.03%,两者联合检测阳性率为87.50%.观察组不同性别之间HCMV感染患儿尿液HCMV-DNA、血清HCMV-IgM及两者联合检测结果均无统计学差异(P＞0.05).肝炎组尿液HCMV-DNA、血清HCMV-IgM及两者联合检出阳性率均高于肺炎组和其他组;且其他组尿液HCMV-DNA、血清HCMV-IgM及两者联合检出阳性率低于肺炎组(P＜0.05).＞180 d尿液HCMV-DNA、血清HCMV-IgM及两者联合检出阳性率均高于＜90 d和90 d-180 d;且90 d-180 d尿液HCMV-DNA、血清HCMV-IgM及两者联合检出阳性率均高于＜90 d(P＜0.05).结论:尿液HCMV-DNA联合血清HCMV-IgM检测在患儿HCMV感染中的诊断效能高,结合临床表现可全面判断患儿HCMV感染状态.     

肝炎综合征婴儿HCMV-IgM定量检测的临床价值

目的：探讨检测肝炎综合征婴儿血清中人巨细胞病毒（human cytomegalovirus,HCMV）IgM含量在诊断和动态监测婴儿HCMV感染中的临床意义。方法：ELISA检测了97例肝炎综合征婴儿（婴肝患儿）血清HCMV-IgM含量,并对其中48例HCMV-IgM阳性患儿在治疗过程中进行了血清HCMV-IgM定量和肝功能的动态监测。结果：97例婴肝患儿血清HCMV-IgM阳性率为49.48%（48/97）,HCMV肝炎患儿治疗前后血清HCMV-IgM定量检测均值分别为（27.7±3.92）、（12.84±2.87）IU/ml,HCMV肝炎患儿治疗后血清HCMV-IgM定量检测值显著下降（t=19.35,P〈0.05）,并与肝脏转氨酶及总胆红素的下降同步。结论：HCMV感染为婴肝患儿的主要病因,血清HCMV-IgM定量检测有助于婴肝患儿的诊断和婴儿巨细胞病毒性肝炎疗效观察。     

荧光定量聚合酶链反应在小儿人巨细胞病毒感染检测中应用

目的探讨荧光定量聚合酶链反应(FQ-PCR)在小儿人巨细胞病毒(HCMV)感染检测中的临床应用价值。方法选择2012年1月至2013年10月在武汉市妇女儿童医疗保健中心就诊的疑似HCMV感染患儿血清或尿液标本,其中血清标本454例,尿液标本1 001例,患儿男女比为1 005∶394,年龄1天~3岁。用FQ-PCR检测疑似HCMV感染患儿血清和尿液中的HCMV-DNA,比较两种标本的阳性率。结果 1 001例尿液标本的HCMV-DNA阳性率为46.35%(464/1 001),显著高于血清标本的阳性率5.95%(27/454);56例血清和尿液标本HCMV-DNA进行配对检测,其中血清检测阴性的52例中有23例尿液检测为阳性,而血清检测阳性的4例尿液检测均为阳性。结论应用FQ-PCR检测患儿HCMV-DNA,采用尿液标本可提高阳性检出率。     

巨细胞病毒感染在抽动障碍中的临床意义初探

目的：探讨人巨细胞病毒（human cytomegalovirus,HCMV)感染在抽动障碍中的临床意义。方法：应用PCR基因扩增技术对66例抽动障碍患儿进行血液HCMV检测,并测定74例正常儿童作为对照,结果：抽动障碍患儿HCMV检出阳性率（26％）明显高于对照组（3％）,差异有显著性（P＜0．01）,抽动障碍三种类型间HCMV感染阳性率无显著性差异（P＞0．05）。结论：HCMV感染与抽动障碍发病有关。     

人巨细胞病毒宫内感染与孕龄的关系

目的研究孕龄与人巨细胞病毒宫内感染的垂直传播率之间的关系。方法分别应用ELISA方法和PCR方法检测273例不同孕龄的孕妇外周血标本中的HCMV-IgM和HCMV-DNA,新生儿血样或胚胎组织用PCR方法检测HCMV-DNA以诊断先天性HCMV感染。结果孕早期妇女外周血中的HCMV-IgM和HCMV-DNA的阳性率均低于孕中期和孕晚期妇女,差异有显著性(P<0.005)。孕中期和孕晚期妇女宫内感染的垂直传播率显著高于孕早期妇女,差异有统计学意义(P<0.001)。结论孕中期和孕晚期妇女发生宫内感染的几率和垂直传播率均显著大于孕早期妇女,应加强对孕中期和孕晚期HCMV感染孕妇的监测和治疗。     

巨细胞病毒感染与儿童免疫性血小板减少性紫癜的关系探讨

目的探讨人巨细胞病毒（HCMV）感染与儿童免疫性血小板减少性紫癜（ITP）的关系。方法采集154例ITP患儿（ITP组）和50例健康儿童（对照组）的血清样本,用Real-time PCR检测HCMV DNA,ELISA方法检测HCMV IgM、IgG抗体;其中105例ITP患儿和50例健康对照儿童采集了尿液标本,用Real-time PCR检测HCMV DNA,并比较ITP组HCMV DNA阳性患儿与阴性患儿的血小板数量的差异。结果 ITP患儿血清HCMV DNA、HCMV IgM及IgG抗体和尿HCMV DNA阳性率均明显高于健康对照儿童,两组比较差异均有统计学意义（P〈0.01）;ITP组HCMV DNA阳性患儿的血小板数量（29.72±14.54）×109/L与阴性患儿（41.28±18.35）×109/L比较差异有统计学意义（P〈0.05）。结论儿童感染HCMV可能是发生ITP的重要致病因素之一,这对指导临床有效治疗及预防ITP的发生有着积极的意义。     

婴儿巨细胞病毒性肝炎患儿体内一氧化氮和内皮型一氧化氮合酶水平的研究CSCD

目的 探讨婴儿巨细胞病毒性肝炎（ICH）患者体内一氧化氮（N0）和内皮型一氧化氮合酶（eNOS）的水平及其临床意义.方法 ELISA方法检测56例ICH患儿和50例健康对照组血清NO和eNOS的水平,统计学方法分析其与患儿肝功能、人巨细胞病毒（HCMV）DNA拷贝数的关联.结果 ICH组NO和eNOS的水平显著高于健康对照组（P〈0.01）.ICH组中ALT异常组NO和eNOS水平显著高于ALT正常组（P〈0.05）.ICH组中NO和eNOS的水平与ALT呈明显正相关（r=0.682,r=0.746,P〈0.05）,但与HCMV-DNA拷贝数无显著相关性（r=-0.087,r=-0.134,P〉0.05）.结论 ICH患儿体内NO以及eNOS水平显著升高,与患儿肝脏功能损伤呈正相关.     

巨细胞病毒宫内感染状态与母婴垂直传播之间的关系

目的研究巨细胞病毒（HCMV）宫内感染状态即活动性与非活动性感染与母婴垂直传播之间的关系。方法对85例HCMV-IgM或HCMV-DNA阳性的母血标本用RT-PCR方法检测即刻早期（IE）mRNA和晚期（L）mRNA,并用PEP-PCR方法检测母血标本中分离出的胎儿细胞的HCMV-DNA。结果 85例母血标本中检测到IE-mRNA或L-mRNA的有57例,其中胎儿细胞HCMV-DNA呈阳性的有37例,宫内垂直传播率为64.91%。未检测到IE-mRNA和L-mRNA的28例样本中胎儿细胞HCMV-DNA呈阳性的有4例,宫内垂直传播率为14.29%。两组的垂直传播率经χ2检验有统计学差异（P〈0.001）。结论巨细胞病毒宫内感染处于活动期状态时,其发生母婴垂直传播的危险性显著高于非活动期感染状态,这对于采取正确的治疗措施和评估预后有重要的指导意义。     

Human cytomegalovirus reactivation during lactation and mother-to-child transmission in preterm infants

In a clinical trial, the incidence of cytomegalovirus reactivation in breastfeeding mothers and transmission to their preterm infants were studied. Breast milk from 73 mothers as well as urine and tracheal and pharyngeal aspirates from their 89 infants were screened weekly for human cytomegalovirus (HCMV) DNA during the first 2 months after delivery. Of the 73 mothers, 48 (66%) were positive for HCMV DNA in the lactating breast. HCMV reactivation could be confirmed for 19 of 20 (95%) immunoglobulin G-positive mothers. Of the eight immunoglobulin G-negative mothers one was positive for HCMV DNA in breast milk. In only 2 out of 13 seropositive mothers with HCMV DNA in breast milk could viral DNA be detected in the peripheral blood. HCMV mother-to-child transmission was concluded for 20 of the 48 (42%) mothers positive for DNA or 7 of 19 (37%) seropositive for HCMV and positive for HCMV DNA in breast milk and one of one mother seronegative for HCMV but positive for HCMV DNA in breast milk. One mother transmitted the virus to her twins. In addition, one infant acquired postnatal HCMV infection despite the mother's being negative for HCMV DNA in breast milk; altogether, we found 22 infants with HCMV infection. In 13 of these 22 infants, virus infection occurred definitively postnatally; two of them developed severe symptomatic HCMV infection. HCMV-infected infants demonstrated higher incidences of amniotic infection, respiratory distress syndrome, bronchopulmonary dysplasia, and retinopathia praenatalis than noninfected infants, however, the differences were not statistically significant. In summary, our study confirmed a very high incidence of HCMV reactivation in mothers during lactation and a significant risk of transmission to preterm infants with the possibility of severe disease in these babies.     

Cytomegalovirus transmission to preterm infants during lactation.

Breastfeeding has a major impact on HCMV epidemiology. The incidence of postnatal HCMV reactivation during lactation equals the maternal seroprevalence. Infectious virus, viral DNA and RNA can be isolated easily from cell and fat-free milk whey. Early onset of viral DNAlactia and virolactia as well as high viral load in milk whey are maternal risk factors for virus transmission. The dynamics of HCMV reactivation can be described by unimodal kinetics with interindividual variation. Virus reactivation during lactation is a self-limiting local process in the absence of systemic HCMV infection. Preterm infants below 1000g birthweight and a gestational age below 30 weeks may be at high risk of acquiring a symptomatic HCMV infection. Several recent studies described low transmission rates and mostly asymptomatically infected neonates using frozen milk. Despite different freeze-storing procedures, HCMV transmissions occurred, and severe HCMV infections were observed. Few data exist on the long-term outcome of postnatally acquired HCMV infection via breast milk. To substantiate the international debate on the use of native or inactivated milk for feeding of preterm infants, additional data are necessary for better identification of mother-infant-pairs at risk for viral transmission and symptomatic infection early after birth.     

Transmission of cytomegalovirus via breast milk to the prematurely born infant: a systematic review

To analyse current data on transmission of human cytomegalovirus (HCMV) via breast milk with subsequent symptomatic HCMV infection of the preterm infant and to report on long-term follow-up, a systematic literature review was performed using EMBASE, MEDLINE and CINAHL (January 1966 to December 2008) Studies were included for analysis if congenital HCMV infection was excluded and transmission via breast milk was either confirmed or strongly suspected. Twenty-six studies were included for analysis. Maternal HCMV-IgG-positivity was reported to be in the range 51.6–100% (median 81.6%), HCMV-IgG detection in breast milk in the range 67–97.2% (median 80%) and HCMV-positivity of the infants in the range 5.7–58.6%. Symptomatic HCMV disease occurred in 0–34.5% (median 3.7%) and severe sepsis-like syndrome in 0–13.8% (median 0.7%). Data on long-term outcome of preterm infants with symptomatic HCMV infection revealed a low risk for mild neurological and cognitive sequelae, without hearing impairment. Recommendations for high-risk preterm infants diverged markedly. The current data report low rates of symptomatic disease after transmission of HCMV via breast milk to the preterm infant without evidence of certain long-term sequelae. The results of our review do not support a general approach, either by avoidance or pasteurization of breast milk, in high-risk preterm infants.     

Detection of human herpesvirus 7 (HHV-7) DNA in breast milk by polymerase chain reaction and prevalence of HHV-7 antibody in breast-fed and bottle-fed children

Twenty-nine breast milk mononuclear cell samples were analyzed for human herpesvirus 7 (HHV-7) DNA, human herpesvirus 6 (HHV-6) DNA, and human cytomegalovirus (HCMV) DNA by polymerase chain reaction (PCR). In addition, peripheral blood mononuclear cell samples from 13 puerperants were analyzed for HHV-7 DNA by PCR, and seropositivity of HHV-7 was also analyzed in breast-fed and bottle-fed children. HHV-7 DNA was detected in 3 of 29 breast milk samples. HCMV DNA was also detected in 3 of 29 breast milk samples, but HHV-6 DNA was not detected. HHV-7 DNA was detected in 11 of 13 samples of peripheral blood mononuclear cells. Though the seropositivity rate for HHV-7 in breast-fed children was slightly higher than that in bottle-fed children at 18 and 24 months old, the difference was not statistically significant. From these results, we speculate that breast-feeding may be one of the transmission routes of HHV-7, although this is not the main route. J. Med. Virol. 56:275–279, 1998. © 1998 Wiley-Liss, Inc.     

Detection of cytomegaloviral DNA in human milk cells and cell free milk whey by nested PCR

Human cytomegalovirus (HCMV) DNA can be detected in different compartments of human milk. A protocol for the preparation of milk whey free of fat and cells for the detection of human cytomegalovirus (HCMV) by nested PCR is presented. This is based upon the experience of the separation of more than 200 milk specimens of healthy seropositive breast feeding mothers. HCMV DNA could be detected in freshly centrifuged and filtrated milk whey specimens without contamination by cellular DNA. In limiting dilution experiments using HCMV plasmid DNA, the effect of different DNA extraction procedures from native milk and milk whey on the detection limit of cytomegaloviral DNA was demonstrated. About 200 viral genome equivalents/ml in milk whey or native milk were detectable by classical organic phenol/chloroform extraction or a spin column method, respectively. The detection of viral DNA in milk cells depended on a minimum number of milk cells (10⁵–2×10⁵) available for DNA extraction. In contrast to the findings of cytomegaloviral DNA in native sera or plasma of immunosuppressed patients we failed to amplify low level viral DNA from native breast milk by nested PCR due to an inhibition of Taq polymerase by lipid components. Finally, the course of cell associated and cell free DNAlactia was monitored. Analyzing sequential milk specimens, in some cases the presence of HCMV DNA in colostrum could be demonstrated. DNAlactia of milk cells and whey was partially discordant. Onset (week 1–4 after delivery) and duration (2 weeks up to more than 3 months) of DNAlactia showed distinct individual patterns. The methods described, allow further analysis of the mechanisms involved in the postnatal HCMV transmission by breast feeding seropositive mothers.     

Human cytomegalovirus glycoprotein B genotypes in Brazilian mothers and their congenitally infected infants

A case-control study design was used in order to compare the distribution of human cytomegalovirus (HCMV) glycoprotein B (gB) genotypes in 48 mothers of 49 congenitally infected infants with that observed in 144 mothers of 146 uninfected infants to study genetic variation of HCMV strains and maternal-fetal transmission. Congenital infection with HCMV was characterized by DNA detection and virus isolation from two urine or saliva samples collected prior to the third week of life. Genotyping of HCMV was carried out by a polymerase chain reaction-restriction fragment length polymorphism analysis of the variable region of the gB gene, testing for four genotypes. Genotype frequency was similar among the 28 non-transmitting mothers who were shedding virus (gB1: 25%; gB2: 28.6%; gB3: 42.8%; gB4: 0%), the 37 transmitting mothers (gB1: 21.6%; gB2: 46%; gB3: 27%; gB4: 0%), and the 49 infected infants (gB1: 39%; gB2: 37%; gB3: 24%; gB4: 0%). The same genotype was detected at different body sites (urine, saliva, and blood) of each infected newborn and in the respective mother (breast milk, urine, and saliva). Co-infection with multiple genotypes was observed in the immediate postpartum period in two mothers of infected infants (5.4%) and one non-transmitting mother (3.6%). The gB genotype was not correlated with intrauterine HCMV transmission. The genotype distribution found reflects the overall frequency of wild strains circulating in this geographic region. A single genotype is responsible for congenital HCMV infection. Co-infection with more than one strain, as characterized by gB genotype, was infrequent in women who were presumably immunocompetent.     

Viral Load in Breast Milk Correlates with Transmission of Human Cytomegalovirus to Preterm Neonates, but Lactoferrin Concentrations Do Not

In vitro, lactoferrin (LF) strongly inhibits human cytomegalovirus (HCMV), which led us to hypothesize that in vivo HCMV might also be inhibited in secretions with high LF concentrations. In breast milk, high viral loads observed as high viral DNA titers tended to coincide with higher LF levels. However, the LF levels did not correlate to virus transmission to preterm infants. The viral load in the transmitting group was highest compared to the nontransmitting group. We conclude that viral load in breast milk is an important factor for transmission of the virus.     

Viral load in breast milk correlates with transmission of human cytomegalovirus to preterm neonates, but lactoferrin concentrations do not.

In vitro, lactoferrin (LF) strongly inhibits human cytomegalovirus (HCMV), which led us to hypothesize that in vivo HCMV might also be inhibited in secretions with high LF concentrations. In breast milk, high viral loads observed as high viral DNA titers tended to coincide with higher LF levels. However, the LF levels did not correlate to virus transmission to preterm infants. The viral load in the transmitting group was highest compared to the nontransmitting group. We conclude that viral load in breast milk is an important factor for transmission of the virus.     

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.