人疱疹病毒6型在儿科临床中的研究进展

近年来,在神经系统疾病患儿中不断检测到人疱疹病毒6型(HHV-6)感染,部分患儿多年以后出现明显的神经系统后遗症,导致生活质量下降.该文简述HHV-6在病毒学和流行病学方面的特征,从原发性感染、中枢神经系统疾病和免疫缺陷病方面对HHV-6感染进行综述.强调了HHV-6感染与儿科中枢神经系统疾病如癫癎、脑炎、脑病的关系以及对HHV-6进行分型的必要性,从而为临床诊断和对HHV-6的进一步研究提供线索.     

人疱疹病毒6型荧光定量分型方法的建立和临床应用

目的 建立对人疱疹病毒6型(HHV-6)能同时进行定量和分型的荧光定量PCR检测新方法,运用该方法对临床疑似病毒性脑炎患儿进行检测.方法 以HHV-6聚合酶基因区(U38)为靶序列,设计通用引物和特异性分型探针,建立能同时检测HHV-6型A/B亚型的荧光定量PCR方法,进行敏感性和特异性实验.对临床445例疑似脑炎患儿的脑脊液标本进行HHV-6荧光定量分型检测,阳性结果测序验证.结果 HHV-6A和HHV-6B病毒株荧光定鼍分型检测结果均为阳性,两亚型之间无交叉.单纯疱疹病毒1型和2型、水痘-带状疱疹病毒、巨细胞病毒、爱泼斯坦.马尔病毒、乙肝病毒、金黄色葡萄球菌、肺炎支原体、人类基因组DNA及空白对照均为阴性.HHV-6荧光定量分型最低能检测到10拷贝/μl HHV-6A/B.在临床445例疑似脑炎患儿脑脊液标本中检出HHV-6阳性21例(4.72%),其中HHV-6A阳性4例,HHV-6B阳性16例,HHV-6A和HHV-6B混合感染1例.整个PCR操作过程2-3 h.结论 HHV-6荧光定量分型方法能对HHV-6同时进行定量和分型,具有特异、敏感、简便、快速的特点,可为临床HHV-6感染性脑炎提供早期、敏感的诊断依据.     

合理开展病原学检测指导精准抗感染治疗

近年来病原学检测技术快速发展, 临床不恰当的使用和解读可导致误诊、误治。合理开展病原学检测要求了解检测方法的特点, 结合病程、病情和感染部位, 病程早期对感染局部样本进行核酸或抗原检测, 病程7 d后可采用血清特异性抗体检测;疑难危重患儿可恰当使用二代测序等技术;可根据感染部位选择不同组合形式的多病原联合检测。规范采集技术, 保证送检样本质量并科学解读检测报告, 方可实现精准抗感染治疗。     

原发性HHV-7感染及其与HHV-6感染的比较

1990年 ,人类疱疹病毒 7型 ( HHV- 7)被首次分离发现。它和 HHV- 6有抗原相关性 ,而关于 HHV- 7原发感染的临床和病毒学特征所知甚少。该文研究目的在于明确儿童原发性 HHV- 7感染的临床和病毒学特征 ,并与原发性 HHV- 6感染的特点作一比较。病例和方法　 1995年 6～ 11月 ,对 496例儿童 (年龄≤ 3岁 )进行观察研究。有发热、中毒症状或伴惊厥等急性疾病表现者 2 5 0例 ,健康儿童 65例 ,有慢性疾病表现者 181例。分离外周血单核细胞 ( PBMCs)并进行培养 ,HHV- 7或 HHV- 6培养阳性用间接荧光抗体染色法鉴定 ,对分离的 PBMCs用套…     

二代测序诊断异基因造血干细胞移植术后合并人类疱疹病毒-6相关脑炎1例报告并文献复习

<正>人类疱疹病毒6型(HHV-6)是幼儿急疹的主要病原体,超过95%的成人在儿童某一阶段均感染过该病毒。原发感染后,病毒潜伏在体内,只有处于免疫抑制状态时病毒才会出现活化~([1-2])。对于接受造血干细胞移植术(HSCT)的病人在术后出现HHV-6的活化是很常见的,临床表现通常无明显症状。近年来,许多学者认为HHV-6的再激活通常发生在造血干细胞移植术后30d内,并与移植相关并发症的发生率和死亡率密切相关。据文献报道,HHV-6脑炎是异基因造血干细胞移植术后发生病毒性脑炎的常见原因,HHV-6脑炎的发生率在脐血     

婴儿 HHV-6感染致暴发性致死性肝炎

人疱疹病毒6型(HHV-6)可引起幼儿急疹,此病通常为婴儿有皮疹的发热性疾病。HHV-6感染亦可表现为没有皮疹而有发热,或有皮疹而无发热。日前已报道的原发性 HHV-6感染的所有疾病,均为良性自限性疾病。本文报道 HHV-6感染引起致死性暴发性肝炎1例。     

百日咳综合征病原学、发病机制与临床特点研究进展

百日咳综合征是由流行性感冒病毒及副流行性感冒病毒、腺病毒、呼吸道合胞病毒、巨细胞病毒、支原体等病原体感染引起的一组临床症候群。百日咳综合征发病率呈周期性上升趋势。本文就百日咳综合征病原学、发病机制与临床特点研究进展展开综述。     

基于症候群检测在感染性疾病实验室病原诊断中的应用

急性感染性疾病仍然是儿科最常见的疾病。准确和及时地作出病原学诊断对于帮助这类疾病的有效治疗至关重要。基于症候群检测的多重核酸分子检测技术是指使用单一实验同时检测多种可造成相似症状的病原的检测方法。因其敏感性、特异性高并且快速高效在近几年被迅速广泛用于呼吸道、消化道、血流、中枢神经系统及关节感染的实验室病原快速诊断。     

新生儿中枢神经系统感染101例临床特征及病原学分析

目的分析新生儿中枢神经系统感染的临床特征及病原学结果, 为精准治疗、优化病原学检测策略提供依据。方法对2020年1月至2021年8月河北省儿童医院新生儿科收治的中枢神经系统感染患儿的临床资料进行数据收集, 分析采用常规及分子生物学技术检出脑脊液病原学情况, 比较不同种类病原体感染的实验室检查特点。结果共纳入101例患儿, 中位出生胎龄为38.8(36.2, 39.6)周, 早产儿27例(26.7%), 男婴68例(67.3%), 中位发病日龄为9(2, 14)d。血培养阳性19例(18.8%), 其中17例细菌, 2例真菌。自脑脊液直接检出病原体者33例(32.7%), 其中细菌13例, 病毒19例, 真菌1例。无乳链球菌、大肠埃希菌是前2位细菌, 检出最常见的病毒是肠道病毒。脑脊液病原体检出方式:细菌培养检出7例(7/101, 6.9%)、脑脊液涂片检出2例(2/21, 9.5%)、单病毒及多重聚合酶链反应方法共检出22例(22/45, 48.9%)、宏基因组二代测序检出4例(4/7, 57.1%)。自脑脊液检出细菌/真菌的患儿脑脊液白细胞计数、蛋白水平以及血C-反应蛋白均较检出病毒...     

泛病原体基因芯片在小儿肺炎中检出博卡病毒

正呼吸道感染是儿童常见病,病原学复杂,20%~30%的小儿呼吸道感染病原难以明确~([1-2])。传统的病原学检测手段如荧光定量PCR、抗原抗体免疫学方法或细菌培养等,操作繁琐、通量低,一次只能检测少数几种致病原~([3-6])。本研究对1例病原不明的小儿肺炎,利用泛病原体基因芯片进行高通量病原学筛查分析,并予后续PCR扩增及测序验证,以探讨该芯片系统应用于呼吸道感染病原学诊断的可行性。     

人类疱疹病毒6型(幼儿急疹)无热性感染

人类疱疹病毒6型(HHV-6)是一种引起幼儿急疹的病原体。该病在婴幼儿中有多种临床表现,包括不显性感染、发热而无皮疹以及发热后伴有皮疹的典型症状。我门报告2例有皮疹而无发热的一种新的婴儿急疹的临床特征。     

儿童2019新型冠状病毒感染的诊断与防治建议(试行第一版)

2019年12月以来,湖北省武汉市暴发新型冠状病毒肺炎流行,疫情很快扩散,形势严峻。随着疫情高峰及更多地区开展病原学检测,儿童感染报告病例数正在明显增多,备受各方关注和高度重视。中华医学会儿科学分会和中华儿科杂志编辑委员会组织相关专业专家在总结部分儿童病例流行病学特征和临床疾病特点的基础上,重点就儿童病例诊断标准、处置流程、治疗和防控等提出建议。     

儿童呼吸道病毒感染的实验室诊断方法及合理应用

病毒是儿童呼吸道感染的主要病原。在呼吸道感染中,早期、快速、准确的病原学诊断,可避免抗生素滥用。呼吸道病毒感染的不同实验室诊断方法各具优缺点,合理运用和正确解读其检测结果有助于临床医生做出准确的病原学诊断。     

儿童呼吸道病毒感染的实验室诊断方法及合理应用

病毒是儿童呼吸道感染的主要病原。在呼吸道感染中,早期、快速、准确的病原学诊断,可避免抗生素滥用。呼吸道病毒感染的不同实验室诊断方法各具优缺点,合理运用和正确解读其检测结果有助于临床医生做出准确的病原学诊断。     

儿童肺炎支原体感染咽拭子核酸检测方法的比较

目的 探讨咽拭子核酸检测方法在儿童肺炎支原体(MP)感染病原学诊断中的应用价值。方法 选取2018年7至10月在浙江大学医学院附属儿童医院呼吸科住院的454例(男210例、女244例)肺炎患儿进行前瞻性研究,入院当天或第2天采集患儿的咽拭子和静脉血,分别采用DNA荧光定量扩增、RNA恒温扩增、MP培养和MP-IgM进行MP病原检测,以MP培养阳性或其余3项检测中2项阳性为MP感染诊断标准并进行二次检测,筛选出MP感染患儿于出院前再次采集咽拭子,采用DNA荧光定量扩增、RNA恒温扩增、MP培养3种方法再次进行MP病原学检测。比较3种检测方法随病情变化的检出率及MP菌量变化,组间比较采用χ2检验。结果 454例住院肺炎患儿采用DNA荧光定量扩增、RNA恒温扩增、MP培养和MP-IgM的检出率分别为43.6%(198/454)、43.2%(196/454)、40.0%(180/454)、30.6%(139/454),差异有统计学意义(χ2=20.8,P<0.05)。DNA荧光定量扩增和RNA恒温扩增检出率差异无统计学意义(χ2=0.018,P=0.900),较MP-IgM及MP培养高。...     

人疱疹病毒6型荧光定量分型方法的建立和临床应用

Objective Human herpesvirus 6(HHV-6)isolates are classified into two variants,HHV-6A and HHV-6B,based on distinct genetic,antigenic and biological characteristics.HHV-6 has been associated with encephalitis in children recently.This study aireed to estabhsh a real time PCR assay for simultaneous detection of the two subtypes of HHV-6,and apply this new assay to children with suspected encephalitis,then analyze the relationship between the infeetion with HHV-6 and encephalitis in children.Method The universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene(U38)of HHV-6.The 5'end of the probes for HHV-6A and HHV-6B was labeled with the fluoreseein reporter tetrachloro-6-carboxyfluorescein and 6-earboxyfluorescein(6-FAM),separately,while the 3'end were quenched with 6-carboxy-tetramethyl-rhedamine.The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established.Then,the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 109 to 10°copies/μl,together with controls were used for testing both sensitivity and specificity of the real time PCR assay.The cerebrospinal fluid(CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.Result Both HHV6A(strain ZJ-159)and HHV-6B (strain GS)were positive on the real time PCR assay.There were no cross-reaction with herpes simplex virus type 1,type 2(HSV-1,HSV-2),varicella-zoster virus(YZV),cytomegalovirus(CMV),EpsteinBarr virus(EBV),hepatitis B virus,Staphylococcus aureus,Mycoplasma pneumoniae and human DNA.A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6.The sensitivity threshold was 10 copies/μl for the real time PCR.HHV-6 positive rate by the real time PCR assay was 4.72%(21/445),including 4 ca8es with HHV-6A infection,16 cases of HHV-6B infeedon and l case with mixed HHV-6A and HHV-6B infeetion.The new PCR assay usually took 2 to 3 hours to provide results.Conclusion This new real time PCR assay call simultaneously detect both subtypes of HHV-6,and have high specificity and sensitivity.It will pmvide an early and sensitive diagnosis of HHV-6 encephalitis in children.     

人疱疹病毒6型荧光定量分型方法的建立和临床应用

Objective Human herpesvirus 6(HHV-6)isolates are classified into two variants,HHV-6A and HHV-6B,based on distinct genetic,antigenic and biological characteristics.HHV-6 has been associated with encephalitis in children recently.This study aireed to estabhsh a real time PCR assay for simultaneous detection of the two subtypes of HHV-6,and apply this new assay to children with suspected encephalitis,then analyze the relationship between the infeetion with HHV-6 and encephalitis in children.Method The universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene(U38)of HHV-6.The 5'end of the probes for HHV-6A and HHV-6B was labeled with the fluoreseein reporter tetrachloro-6-carboxyfluorescein and 6-earboxyfluorescein(6-FAM),separately,while the 3'end were quenched with 6-carboxy-tetramethyl-rhedamine.The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established.Then,the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 109 to 10°copies/μl,together with controls were used for testing both sensitivity and specificity of the real time PCR assay.The cerebrospinal fluid(CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.Result Both HHV6A(strain ZJ-159)and HHV-6B (strain GS)were positive on the real time PCR assay.There were no cross-reaction with herpes simplex virus type 1,type 2(HSV-1,HSV-2),varicella-zoster virus(YZV),cytomegalovirus(CMV),EpsteinBarr virus(EBV),hepatitis B virus,Staphylococcus aureus,Mycoplasma pneumoniae and human DNA.A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6.The sensitivity threshold was 10 copies/μl for the real time PCR.HHV-6 positive rate by the real time PCR assay was 4.72%(21/445),including 4 ca8es with HHV-6A infection,16 cases of HHV-6B infeedon and l case with mixed HHV-6A and HHV-6B infeetion.The new PCR assay usually took 2 to 3 hours to provide results.Conclusion This new real time PCR assay call simultaneously detect both subtypes of HHV-6,and have high specificity and sensitivity.It will pmvide an early and sensitive diagnosis of HHV-6 encephalitis in children.     

人疱疹病毒6型荧光定量分型方法的建立和临床应用

Objective Human herpesvirus 6(HHV-6)isolates are classified into two variants,HHV-6A and HHV-6B,based on distinct genetic,antigenic and biological characteristics.HHV-6 has been associated with encephalitis in children recently.This study aireed to estabhsh a real time PCR assay for simultaneous detection of the two subtypes of HHV-6,and apply this new assay to children with suspected encephalitis,then analyze the relationship between the infeetion with HHV-6 and encephalitis in children.Method The universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene(U38)of HHV-6.The 5'end of the probes for HHV-6A and HHV-6B was labeled with the fluoreseein reporter tetrachloro-6-carboxyfluorescein and 6-earboxyfluorescein(6-FAM),separately,while the 3'end were quenched with 6-carboxy-tetramethyl-rhedamine.The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established.Then,the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 109 to 10°copies/μl,together with controls were used for testing both sensitivity and specificity of the real time PCR assay.The cerebrospinal fluid(CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.Result Both HHV6A(strain ZJ-159)and HHV-6B (strain GS)were positive on the real time PCR assay.There were no cross-reaction with herpes simplex virus type 1,type 2(HSV-1,HSV-2),varicella-zoster virus(YZV),cytomegalovirus(CMV),EpsteinBarr virus(EBV),hepatitis B virus,Staphylococcus aureus,Mycoplasma pneumoniae and human DNA.A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6.The sensitivity threshold was 10 copies/μl for the real time PCR.HHV-6 positive rate by the real time PCR assay was 4.72%(21/445),including 4 ca8es with HHV-6A infection,16 cases of HHV-6B infeedon and l case with mixed HHV-6A and HHV-6B infeetion.The new PCR assay usually took 2 to 3 hours to provide results.Conclusion This new real time PCR assay call simultaneously detect both subtypes of HHV-6,and have high specificity and sensitivity.It will pmvide an early and sensitive diagnosis of HHV-6 encephalitis in children.     

人疱疹病毒6型荧光定量分型方法的建立和临床应用

Objective Human herpesvirus 6(HHV-6)isolates are classified into two variants,HHV-6A and HHV-6B,based on distinct genetic,antigenic and biological characteristics.HHV-6 has been associated with encephalitis in children recently.This study aireed to estabhsh a real time PCR assay for simultaneous detection of the two subtypes of HHV-6,and apply this new assay to children with suspected encephalitis,then analyze the relationship between the infeetion with HHV-6 and encephalitis in children.Method The universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene(U38)of HHV-6.The 5'end of the probes for HHV-6A and HHV-6B was labeled with the fluoreseein reporter tetrachloro-6-carboxyfluorescein and 6-earboxyfluorescein(6-FAM),separately,while the 3'end were quenched with 6-carboxy-tetramethyl-rhedamine.The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established.Then,the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 109 to 10°copies/μl,together with controls were used for testing both sensitivity and specificity of the real time PCR assay.The cerebrospinal fluid(CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.Result Both HHV6A(strain ZJ-159)and HHV-6B (strain GS)were positive on the real time PCR assay.There were no cross-reaction with herpes simplex virus type 1,type 2(HSV-1,HSV-2),varicella-zoster virus(YZV),cytomegalovirus(CMV),EpsteinBarr virus(EBV),hepatitis B virus,Staphylococcus aureus,Mycoplasma pneumoniae and human DNA.A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6.The sensitivity threshold was 10 copies/μl for the real time PCR.HHV-6 positive rate by the real time PCR assay was 4.72%(21/445),including 4 ca8es with HHV-6A infection,16 cases of HHV-6B infeedon and l case with mixed HHV-6A and HHV-6B infeetion.The new PCR assay usually took 2 to 3 hours to provide results.Conclusion This new real time PCR assay call simultaneously detect both subtypes of HHV-6,and have high specificity and sensitivity.It will pmvide an early and sensitive diagnosis of HHV-6 encephalitis in children.     

人疱疹病毒6型荧光定量分型方法的建立和临床应用

Objective Human herpesvirus 6(HHV-6)isolates are classified into two variants,HHV-6A and HHV-6B,based on distinct genetic,antigenic and biological characteristics.HHV-6 has been associated with encephalitis in children recently.This study aireed to estabhsh a real time PCR assay for simultaneous detection of the two subtypes of HHV-6,and apply this new assay to children with suspected encephalitis,then analyze the relationship between the infeetion with HHV-6 and encephalitis in children.Method The universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene(U38)of HHV-6.The 5'end of the probes for HHV-6A and HHV-6B was labeled with the fluoreseein reporter tetrachloro-6-carboxyfluorescein and 6-earboxyfluorescein(6-FAM),separately,while the 3'end were quenched with 6-carboxy-tetramethyl-rhedamine.The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established.Then,the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 109 to 10°copies/μl,together with controls were used for testing both sensitivity and specificity of the real time PCR assay.The cerebrospinal fluid(CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.Result Both HHV6A(strain ZJ-159)and HHV-6B (strain GS)were positive on the real time PCR assay.There were no cross-reaction with herpes simplex virus type 1,type 2(HSV-1,HSV-2),varicella-zoster virus(YZV),cytomegalovirus(CMV),EpsteinBarr virus(EBV),hepatitis B virus,Staphylococcus aureus,Mycoplasma pneumoniae and human DNA.A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6.The sensitivity threshold was 10 copies/μl for the real time PCR.HHV-6 positive rate by the real time PCR assay was 4.72%(21/445),including 4 ca8es with HHV-6A infection,16 cases of HHV-6B infeedon and l case with mixed HHV-6A and HHV-6B infeetion.The new PCR assay usually took 2 to 3 hours to provide results.Conclusion This new real time PCR assay call simultaneously detect both subtypes of HHV-6,and have high specificity and sensitivity.It will pmvide an early and sensitive diagnosis of HHV-6 encephalitis in children.