CAJ | 학술논문

目的了解疑似中枢神经系统(CNS)感染患儿中人副肠孤病毒(HPeV)的感染状况及脓毒症感染的关系.方法回顾性收集2012年因疑似CNS感染而在首都儿科研究所附属儿童医院住院患儿的脑脊液标本.采用针对HPeV基因组相对保守的5'端非编码区和变异显著的衣壳蛋白基因3和1交界区的通用引物和分型引物,分别对脑脊液标本核酸进行反转录巢式PCR (RT-nPCR)扩增.阳性标本的PCR产物直接进行核苷酸序列测定,与GenBank中的序列进行比对分析并建立系统进化树.对HPeV筛查阳性的脑脊液标本进行肠道病毒(EV)、EB病毒(EBV)和人巨细胞病毒(HCMV)的检测.同时收集HPeV筛查阳性患儿的其他类型标本(包括血清、鼻咽洗液、粪便标本等)进行上述检测.结果 577份脑脊液标本中检测到18份HPeV阳性标本,检出率为3.1％.阳性患儿中男12例,女6例,年龄范围为15 d～14岁,其中3个月以下患儿最多,占7例;其次是3个月～1岁年龄组5例;另有3例阳性患儿＞9岁.对分型成功的10份脑脊液标本中HPeV的序列分析显示7份为HPeV 3型,3份为HPeV 1型.18份HPeV筛查阳性的脑脊液标本中EV、EBV和HCMV检测均为阴性.脑脊液HPeV筛查阳性患儿的其他类型标本中,8份血清标本中检出2份阳性(均为HPeV3),其中1份与脑脊液标本检出及分型结果一致,而另1份对应脑脊液标本筛查为阳性,但未分出型别;2份粪便标本中检出1份阳性(HPeV1),但对应脑脊液标本未分出型别.HPeV阳性患儿临床诊断主要是脓毒症(7/18)和CNS感染(4/18).其中6例(6/8) HPeV3阳性患儿诊断为脓毒症.HPeV3感染多集中于8月份,而HPeV1则集中于1月份.结论 HPeV与北京儿童脓毒症和CNS感染性疾病相关.1岁以下儿童为易感人群.HPeV3型为脑脊液中主要的检出型别.
목적 료해의사중추신경계통(CNS)감염환인중인부장고병독(HPeV)적감염상황급농독증감염적관계.방법 회고성수집2012년인의사CNS감염이재수도인과연구소부속인동의원주원환인적뇌척액표본.채용침대HPeV기인조상대보수적5'단비편마구화변이현저적의각단백기인3화1교계구적통용인물화분형인물,분별대뇌척액표본핵산진행반전록소식PCR (RT-nPCR)확증.양성표본적PCR산물직접진행핵감산서렬측정,여GenBank중적서렬진행비대분석병건립계통진화수.대HPeV사사양성적뇌척액표본진행장도병독(EV)、EB병독(EBV)화인거세포병독(HCMV)적검측.동시수집HPeV사사양성환인적기타류형표본(포괄혈청、비인세액、분편표본등)진행상술검측.결과 577빈뇌척액표본중검측도18빈HPeV양성표본,검출솔위3.1％.양성환인중남12례,녀6례,년령범위위15 d～14세,기중3개월이하환인최다,점7례;기차시3개월～1세년령조5례;령유3례양성환인＞9세.대분형성공적10빈뇌척액표본중HPeV적서렬분석현시7빈위HPeV 3형,3빈위HPeV 1형.18빈HPeV사사양성적뇌척액표본중EV、EBV화HCMV검측균위음성.뇌척액HPeV사사양성환인적기타류형표본중,8빈혈청표본중검출2빈양성(균위HPeV3),기중1빈여뇌척액표본검출급분형결과일치,이령1빈대응뇌척액표본사사위양성,단미분출형별;2빈분편표본중검출1빈양성(HPeV1),단대응뇌척액표본미분출형별.HPeV양성환인림상진단주요시농독증(7/18)화CNS감염(4/18).기중6례(6/8) HPeV3양성환인진단위농독증.HPeV3감염다집중우8월빈,이HPeV1칙집중우1월빈.결론 HPeV여북경인동농독증화CNS감염성질병상관.1세이하인동위역감인군.HPeV3형위뇌척액중주요적검출형별.
Objective Human parechovirus (HPeV) is a single-stranded,positive sense RNA virus in the Parechovirus genus within the large family of Picornaviridae.As a possible new pathogen of neonatal sepsis,meningoencephalitis and other infections in young children,HPeV gets more and more attention.This study aimed to better understand the association of HPeV with central nervous system (CNS) infectious diseases and sepsis among hospitalized children in Beijing.Method A total of 577 cerebrospinal fluid (CSF) samples were retrospectively collected from 557 children suspected of CNS infections in 2012.Three hundred and fifty-one of them were male and 206 were female.HPeV was screened by reverse transcription-nested PCR (RT-nPCR) with the universal primers which target the highly conserved 5'UTR.The positive samples were genotyped by amplifying and sequencing for the VP3/VP1 junction region.The sequences were compared with the HPeV sequences from GenBank and performed phylogenetic analysis.Some samples other than CSF from HPeV positive children,including serum,nasopharyngeal aspirate and stool,were collected and carried out screening for HPeV.Result With the RT-nPCR by universal primers,HPeVs were detected in 18 out of 577 CSF samples obtained from 18 children with a positive rate of 3.1％.The ratio of male and female was 2 ∶ 1.There were no statistically significant differences on infection rate between boys (12/351,3.4％) and girls (6/206,2.9％).All of 18 positive CSF samples were negative for enterovirus,Epstein-Barr virus (EBV),human cytomegalovirus (HCMV),and herpes simplex virus 1 and 2 (HSV).HPeVs from 10 positive CSF samples were genotyped successfully,consisting of 7 HPeV3 and 3 HPeV1.In addition,2 of 8 serum samples were positive for HPeV3 and 1 of 2 stool samples were positive for HPeV 1.HPeVs were identified in CSF from children aged from 15 days to 14 years,in which 7 cases were infants younger than 3 months and 5 cases were infants from 3 months to one year.Three children older than the age of 9 years (9,13 and 14 years) were positive for HPeV.Most of the children (6/8) infected with HPeV3 were younger than 3 months and were diagnosed as sepsis,while the rest of HPeV3 positive children were diagnosed as meningitis and bronchopneumonia.HPeV3 infection clustered in August,while HPeV1 in January.Conclusion HPeVs were associated with CNS infections and sepsis in hospitalized children in Beijing,especially in children younger than one year.HPeV3 was the predominant type identified in CSF.