北京协和医院就诊儿童EB病毒血清流行病学研究
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摘要
EB病毒感染与许多肿瘤的发生密切相关。目前,我国缺乏关于人群EB病毒感染,尤其是儿童初次感染的血清流行病学资料；而了解儿童感染EB病毒的概貌,对于病毒学基础研究、EB病毒相关疾病的治疗和预防等,都具有参考意义。
本研究以2013年1月-3月在北京协和医院就诊,且进行过任意一种血清学检查的0-9岁儿童作为研究样本,通过酶联免疫吸附试验(ELISA)的方法检测EB病毒的4种标记性抗体,即抗VCA (viral capsid antigen) IgM及IgG抗体、抗EA(early antigen) IgG抗体和抗NA(nuclear antigen) IgG抗体,所测抗体的结果构成了研究样本的抗体谱。这种联合检测抗EB病毒抗体谱的实验方法在国内并不多见,在北京地区则为首次采用。由抗体谱的结果将所有研究样本区分为急性感染、既往感染和未感染三大类,按照年龄分布,统计了各年龄组的阳性率。除此以外,本研究还提取了研究样本的血清游离DNA,通过实时荧光定量PCR的方法检测了EB病毒DNA的阳性率和阳性标本的DNA拷贝数。
研究结果显示,在所有754份符合设定要求的研究样本中,有583例(80.1%)样本的抗体谱提示EB病毒感染,其中急性感染和既往感染分别有65例(8.92%)和518例(71.2%)。在年龄分布上,根据抗VCA IgM抗体的结果(该抗体不会经过胎盘从母体进入婴儿体内),并结合DNA检测的结果,0岁(0-6m)的感染阳性率为0;根据4种抗体的抗体谱结果,从0岁(7-11m,42.3%)起到9岁(91.9%)的感染阳性率呈现逐渐升高的趋势,其中从0岁(7-11m)到2岁是增长率最快的阶段；而感染阳性率在8岁年龄组首次超过900%,达到与成年人相当的水平。参考以往类似的研究资料,将本研究提示的感染阳性率与北京地区20世纪70年代及2006年的数据结合分析,其感染高峰呈现出延后的趋势,逐渐向发达国家的感染高峰靠近,与我国现阶段的经济社会发展水平相符合。
在754份研究样本中,血清病毒DNA阳性的样本为39例,占5.17%,明显低于抗体谱的阳性率。由于血清EB病毒DNA仅在病毒活跃复制时才能检测到,使用实时荧光定量PCR的方法检测血清病毒DNA拷贝数更适合于在具有一定感染风险的患儿中进行EB病毒感染相关疾病的诊断或评价疾病的严重性时使用。在以无症状的病毒携带者为主要人群的正常人中进行筛查,其对病毒感染状态的提示意义是较为有限的。
EBV is associated with a number of malignancies. Seroepidemiological data concerning children's primary infection of EBV in China is rare, yet such information would help in virology research as well as prevention and treatment of EBV-related diseases.
The study enrolled all children from0-9who visited Peking Union Medical College Hospital between January and March2013and whose serum has been collected for laboratory tests. Four antibodies, namely anti-VCA (viral capsid antigen) IgM and IgG antibodies, anti-EA(early antigen) IgG antibody and an ti-NA (nuclear antigen) IgG antibody, were investigated using ELISA, which was the first such study conducted in Beijing area. According to the results of four different antibodies, the children enrolled in the study are further grouped as primary infection, past infection or susceptible to EBV, and the percentage of each group is calculated. Serum DNA of these children is extracted to investigate EBV DNA positivity as well as copy number, using Real-time PCR technique.
As the result shows,583(80.1%) of the754children in the study are infected with EBV,65(8.92) of whom are in the state of primary infection. The seropositivity of anti-VCA IgM antibodies and EBV DNA positivity revealed that all children of age group0(0-6m) are susceptible to EBV, and starting from the next age group, results of the four antibodies demonstrate a steady increase from42.3%(7-11m) to91.9%(9-year-olds) in seropositivity. The growth rate peaks between age group0(7-11m) and2, and the seropositivity of EBV exceeds90%at age8, which is approximately the positivity of adults. Compared with the results of similar investigations conducted in1970's and2006in Beijing, the age of primary infection is delayed. Since primary infection of EBV tend to occur later in a more developed society, the result coincides with the social-economic development of China in recent decades. Real-time PCR test finds39(5.17%) of the754children are positive for EBV DNA in their serum. As serum EBV DNA is only detectable when the virus undergoes active replication, the determination of serum DNA copies would be more valuable in the diagnosis of patients who are suspected of EBV infection, or in the evaluation of disease severity of such patients. Screening of serum EBV DNA in healthy people or EBV carriers with no symptoms contributes little to determining the status of infection.
本研究以2013年1月-3月在北京协和医院就诊,且进行过任意一种血清学检查的0-9岁儿童作为研究样本,通过酶联免疫吸附试验(ELISA)的方法检测EB病毒的4种标记性抗体,即抗VCA (viral capsid antigen) IgM及IgG抗体、抗EA(early antigen) IgG抗体和抗NA(nuclear antigen) IgG抗体,所测抗体的结果构成了研究样本的抗体谱。这种联合检测抗EB病毒抗体谱的实验方法在国内并不多见,在北京地区则为首次采用。由抗体谱的结果将所有研究样本区分为急性感染、既往感染和未感染三大类,按照年龄分布,统计了各年龄组的阳性率。除此以外,本研究还提取了研究样本的血清游离DNA,通过实时荧光定量PCR的方法检测了EB病毒DNA的阳性率和阳性标本的DNA拷贝数。
研究结果显示,在所有754份符合设定要求的研究样本中,有583例(80.1%)样本的抗体谱提示EB病毒感染,其中急性感染和既往感染分别有65例(8.92%)和518例(71.2%)。在年龄分布上,根据抗VCA IgM抗体的结果(该抗体不会经过胎盘从母体进入婴儿体内),并结合DNA检测的结果,0岁(0-6m)的感染阳性率为0;根据4种抗体的抗体谱结果,从0岁(7-11m,42.3%)起到9岁(91.9%)的感染阳性率呈现逐渐升高的趋势,其中从0岁(7-11m)到2岁是增长率最快的阶段；而感染阳性率在8岁年龄组首次超过900%,达到与成年人相当的水平。参考以往类似的研究资料,将本研究提示的感染阳性率与北京地区20世纪70年代及2006年的数据结合分析,其感染高峰呈现出延后的趋势,逐渐向发达国家的感染高峰靠近,与我国现阶段的经济社会发展水平相符合。
在754份研究样本中,血清病毒DNA阳性的样本为39例,占5.17%,明显低于抗体谱的阳性率。由于血清EB病毒DNA仅在病毒活跃复制时才能检测到,使用实时荧光定量PCR的方法检测血清病毒DNA拷贝数更适合于在具有一定感染风险的患儿中进行EB病毒感染相关疾病的诊断或评价疾病的严重性时使用。在以无症状的病毒携带者为主要人群的正常人中进行筛查,其对病毒感染状态的提示意义是较为有限的。
EBV is associated with a number of malignancies. Seroepidemiological data concerning children's primary infection of EBV in China is rare, yet such information would help in virology research as well as prevention and treatment of EBV-related diseases.
The study enrolled all children from0-9who visited Peking Union Medical College Hospital between January and March2013and whose serum has been collected for laboratory tests. Four antibodies, namely anti-VCA (viral capsid antigen) IgM and IgG antibodies, anti-EA(early antigen) IgG antibody and an ti-NA (nuclear antigen) IgG antibody, were investigated using ELISA, which was the first such study conducted in Beijing area. According to the results of four different antibodies, the children enrolled in the study are further grouped as primary infection, past infection or susceptible to EBV, and the percentage of each group is calculated. Serum DNA of these children is extracted to investigate EBV DNA positivity as well as copy number, using Real-time PCR technique.
As the result shows,583(80.1%) of the754children in the study are infected with EBV,65(8.92) of whom are in the state of primary infection. The seropositivity of anti-VCA IgM antibodies and EBV DNA positivity revealed that all children of age group0(0-6m) are susceptible to EBV, and starting from the next age group, results of the four antibodies demonstrate a steady increase from42.3%(7-11m) to91.9%(9-year-olds) in seropositivity. The growth rate peaks between age group0(7-11m) and2, and the seropositivity of EBV exceeds90%at age8, which is approximately the positivity of adults. Compared with the results of similar investigations conducted in1970's and2006in Beijing, the age of primary infection is delayed. Since primary infection of EBV tend to occur later in a more developed society, the result coincides with the social-economic development of China in recent decades. Real-time PCR test finds39(5.17%) of the754children are positive for EBV DNA in their serum. As serum EBV DNA is only detectable when the virus undergoes active replication, the determination of serum DNA copies would be more valuable in the diagnosis of patients who are suspected of EBV infection, or in the evaluation of disease severity of such patients. Screening of serum EBV DNA in healthy people or EBV carriers with no symptoms contributes little to determining the status of infection.
引文
[1]曾益新.肿瘤学第3版[M].北京：人民卫生出版社,2012：23.
[2]Rickinson A, Kieff E. Epstein-Barr virus. In:Knipe D, Howley P, eds. Fields Virology, 4th ed[M]. Philadelphia:Lippincott Williams & Wilkins,2001:2575-627.
[3]Hesse J, Ibsen K K, Krabbe S, et al. Prevalence of antibodies to Epstein-Barr virus (EBV) in childhood and adolescence in DenmarkQ]. Scandinavian journal of infectious diseases,1983, 15(4):335.
[4]Chan K H, Tam J S L, Peiris J S M, et al. Epstein-Barr virus (EBV) infection in infancy[J]. Journal of clinical virology,2001,21(1):57-62.
[5]Kangro H O, Osman H K, Lau Y L, et al. Seroprevalence of antibodies to human herpesviruses in England and Hong Kong[J]. Journal of medical virology,1994,43(1):91-96.
[6]Niederman J C, Evans A S, Subrahmanyan L, et al. Prevalence, incidence and persistence of EB virus antibody in young adults [J]. New England Journal of Medicine,1970,282(7):361-5.
[7]de-The G. Epidemiology of Epstein-Barr virus and associated diseases in man. [A] In: Roizman B, editor. The Herpesviruses, vol. 1.[M] New York:Plenum Publishing Corp, 1982:25-103.
[8]杜海军,周玲,刘宏图,等.北京地区儿童EB病毒感染的血清学调查[J].中华实验和临床病毒学杂志,2008,22(1)：30-32.
[9]Bauer C C, Aberle S W, Popow-Kraupp T, et al. Serum Epstein-Barr virus DNA load in primary Epstein-Barr virus infection[J]. Journal of medical virology,2005,75(1):54-58.
[10]Kimura H, Ito Y, Suzuki R, et al. Measuring Epstein-Barr virus (EBV) load:the significance and application for each EBV-associated disease[J]. Reviews in medical virology, 2008,18(5):305-319.
[11]柳文菊,杜昆,刘学政,等.儿童呼吸道EB病毒感染IgM抗体与病毒DNA的相关性分析[J].国际检验医学杂志ISTIC,2012,33(3).
[12]H Sousa, J Silva, L Azevedo, et al. Epstein-Barr Virus in healthy individuals from Portugal [J]. Acta Med Port,2011; 24:707-712
[13]Hess R D. Routine Epstein-Barr virus diagnostics from the laboratory perspective:still challenging after 35 years[J]. Journal of clinical microbiology,2004,42(8):3381-3387.
[14]Takeuchi K, Tanaka-Taya K, Kazuyama Y, et al. Prevalence of Epstein-Barr virus in Japan: trends and future prediction [J]. Pathology international,2006,56(3):112-116.
[15]Ito Y, Ishimoto A, Hosokawa T, Hotta S, Noerjasin B. Anti-EB virus antibody titer in sera collected from human subjects of Surabaja, Indonesia, in 1968Q]. Kobe J Med Sci 1970; 16:261-5.
[16]Tsai W S, Chang M H, Chen J Y, et al. Seroepidemiological study of Epstein-Barr virus infection in children in Taipei[J]. Zhonghua Minguo xiao er ke yi xue hui za zhi [Journal]. Zhonghua Minguo xiao er ke yi xue hui,1989,30(2):81.
[17]de-The G, Day NE, Geser A et al. Sero-epidemiology of the Epstein-Barr virus: Preliminary analysis of an international study:A review[J]. IARC Sci Publ 1975; 11:3-16.
[18]Leogrande G, Jirillo E. Studies on the epidemiology of child infections in the Bari area (South Italy) VII. Epidemiology of Epstein-Barr virus infections [J]. European journal of epidemiology,1993,9(4):368-372.
[19]Venkitaraman A R, Lenoir G M, John T J. The seroepidemiology of infection due to Epstein-Barr virus in southern India[J]. Journal of medical virology,1985,15(1):11-16.
[20]李玉雨,郑承勋,李英信,等.延边地区儿童EBV抗体血清流行病学研究[J].延边医学院学报,1991,1：14.
[21]Sumaya C V. Endogenous reactivation of Epstein-Barr virus infections[J]. Journal of Infectious Diseases,1977,135(3):374-379.
[22]陈倩,胡正,张其华.761例住院儿童EB病毒感染分析[J].中国当代儿科杂志,2013,15(3)：183-186.
[23]Fafi-Kremer S, Morand P, Brion J P, et al. Long-term shedding of infectious Epstein-Barr virus after infectious mononucleosis[J]. Journal of Infectious Diseases,2005,191 (6):985-989.
[24]Vasef M A, Ferlito A, Weiss L M. Nasopharyngeal carcinoma, with emphasis on its relationship to Epstein-Barr virusQ]. The Annals of otology, rhinology & laryngology,1997, 106(4):348-356.
[2]Rickinson A, Kieff E. Epstein-Barr virus. In:Knipe D, Howley P, eds. Fields Virology, 4th ed[M]. Philadelphia:Lippincott Williams & Wilkins,2001:2575-627.
[3]Hesse J, Ibsen K K, Krabbe S, et al. Prevalence of antibodies to Epstein-Barr virus (EBV) in childhood and adolescence in DenmarkQ]. Scandinavian journal of infectious diseases,1983, 15(4):335.
[4]Chan K H, Tam J S L, Peiris J S M, et al. Epstein-Barr virus (EBV) infection in infancy[J]. Journal of clinical virology,2001,21(1):57-62.
[5]Kangro H O, Osman H K, Lau Y L, et al. Seroprevalence of antibodies to human herpesviruses in England and Hong Kong[J]. Journal of medical virology,1994,43(1):91-96.
[6]Niederman J C, Evans A S, Subrahmanyan L, et al. Prevalence, incidence and persistence of EB virus antibody in young adults [J]. New England Journal of Medicine,1970,282(7):361-5.
[7]de-The G. Epidemiology of Epstein-Barr virus and associated diseases in man. [A] In: Roizman B, editor. The Herpesviruses, vol. 1.[M] New York:Plenum Publishing Corp, 1982:25-103.
[8]杜海军,周玲,刘宏图,等.北京地区儿童EB病毒感染的血清学调查[J].中华实验和临床病毒学杂志,2008,22(1)：30-32.
[9]Bauer C C, Aberle S W, Popow-Kraupp T, et al. Serum Epstein-Barr virus DNA load in primary Epstein-Barr virus infection[J]. Journal of medical virology,2005,75(1):54-58.
[10]Kimura H, Ito Y, Suzuki R, et al. Measuring Epstein-Barr virus (EBV) load:the significance and application for each EBV-associated disease[J]. Reviews in medical virology, 2008,18(5):305-319.
[11]柳文菊,杜昆,刘学政,等.儿童呼吸道EB病毒感染IgM抗体与病毒DNA的相关性分析[J].国际检验医学杂志ISTIC,2012,33(3).
[12]H Sousa, J Silva, L Azevedo, et al. Epstein-Barr Virus in healthy individuals from Portugal [J]. Acta Med Port,2011; 24:707-712
[13]Hess R D. Routine Epstein-Barr virus diagnostics from the laboratory perspective:still challenging after 35 years[J]. Journal of clinical microbiology,2004,42(8):3381-3387.
[14]Takeuchi K, Tanaka-Taya K, Kazuyama Y, et al. Prevalence of Epstein-Barr virus in Japan: trends and future prediction [J]. Pathology international,2006,56(3):112-116.
[15]Ito Y, Ishimoto A, Hosokawa T, Hotta S, Noerjasin B. Anti-EB virus antibody titer in sera collected from human subjects of Surabaja, Indonesia, in 1968Q]. Kobe J Med Sci 1970; 16:261-5.
[16]Tsai W S, Chang M H, Chen J Y, et al. Seroepidemiological study of Epstein-Barr virus infection in children in Taipei[J]. Zhonghua Minguo xiao er ke yi xue hui za zhi [Journal]. Zhonghua Minguo xiao er ke yi xue hui,1989,30(2):81.
[17]de-The G, Day NE, Geser A et al. Sero-epidemiology of the Epstein-Barr virus: Preliminary analysis of an international study:A review[J]. IARC Sci Publ 1975; 11:3-16.
[18]Leogrande G, Jirillo E. Studies on the epidemiology of child infections in the Bari area (South Italy) VII. Epidemiology of Epstein-Barr virus infections [J]. European journal of epidemiology,1993,9(4):368-372.
[19]Venkitaraman A R, Lenoir G M, John T J. The seroepidemiology of infection due to Epstein-Barr virus in southern India[J]. Journal of medical virology,1985,15(1):11-16.
[20]李玉雨,郑承勋,李英信,等.延边地区儿童EBV抗体血清流行病学研究[J].延边医学院学报,1991,1：14.
[21]Sumaya C V. Endogenous reactivation of Epstein-Barr virus infections[J]. Journal of Infectious Diseases,1977,135(3):374-379.
[22]陈倩,胡正,张其华.761例住院儿童EB病毒感染分析[J].中国当代儿科杂志,2013,15(3)：183-186.
[23]Fafi-Kremer S, Morand P, Brion J P, et al. Long-term shedding of infectious Epstein-Barr virus after infectious mononucleosis[J]. Journal of Infectious Diseases,2005,191 (6):985-989.
[24]Vasef M A, Ferlito A, Weiss L M. Nasopharyngeal carcinoma, with emphasis on its relationship to Epstein-Barr virusQ]. The Annals of otology, rhinology & laryngology,1997, 106(4):348-356.