乙型肝炎病毒父婴垂直传播的危险因素及基因型研究
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摘要
目的
探讨乙型肝炎病毒(HBV)父婴垂直传播的危险因素;分析血液HBV-DNA载量、精液HBV-DNA载量与父婴垂直传播的关联;研究HBV父婴垂直传播中父婴所携HBV的基因型是否一致。
方法
选择2009年8月至2010年12月在福建省妇幼保健院行产前初诊检查的孕妇,问诊了解其HBsAg且HBV-DNA阴性或仅HBsAb阳性、丈夫HBsAg阳性并决定住本院分娩的孕妇家庭;夫妻双方知情同意参与有关血液或精液乙型肝炎标志物(HBVM)及HBV-DNA定量和基因型检测。分娩期再次抽血确定:孕妇HBVM全阴性或仅HBsAb阳性且HBV-DNA阴性、丈夫HBsAg阳性及其新生儿,共154个研究家庭。采用酶联免疫吸附法(ELISA)检测HBVM,荧光定量PCR法(FQ-PCR)检测血液和精液HBV-DNA载量,型特异性引物和高分辨率溶解曲线测定乙肝病毒基因型。
结果
1.新生儿脐血HBV-DNA阳性率为19.5%(30/154),经单因素分析,孕妇HBsAb阳性、父亲血液HBeAg阳性、父亲血液HBV-DNA阳性、精液HBV-DNA阳性、HBV携带时间、新生儿性别有统计学意义(P<0.05);经多因素分析,父亲血液HBeAg阳性、父亲血液HBV-DNA阳性、精液HBV-DNA阳性仍有统计学意义(P<0.05)。
2.脐血HBV-DNA载量与父亲血液HBV-DNA载量呈正相关。
3.抽血孕周与父亲血液HBV-DNA载量相关性小。
4.精液HBV-DNA阳性率为37.5%(30/80);精液HBV-DNA载量与其血液的HBV-DNA载量呈正相关,精液载量低于血液载量。
5.脐血HBVM检测HBsAg阳性率为17.5%(27/154)。
6.精液HBV-DNA载量与采集孕周相关性不大。
7.HBV-DNA载量由高到低依次为:父亲血液HBV-DNA载量>精液HBV-DNA载量>脐血HBV-DNA载量。
8.父亲血液HBV-DNA阳性伴有HBeAg阳性未见交互作用(P>0.05);父亲血液HBV-DNA阳性伴有精液阳性有交互作用(P<0.05)。
9.两组孕妇孕产史和新生儿妊娠结局等情况类似,均无统计学意义(P均>0.05)。
10.父婴HBV-DNA基因型一致,以B型为主,还有少量的C型、D型和B+C混合型。
11.C型的HBV-DNA载量高于其他型。
12.福州市区和郊县的HBV父婴垂直传播率类似(P>0.05)。
13.阳性新生儿随访至第6个月时,阴转率为6.25%(1/16)。
结论
1.父亲血液HBV-DNA阳性、精液HBV-DNA阳性、血液HBeAg阳性是HBV父婴垂直传播的危险因素。
2.父亲血液HBV-DNA载量越高,其精液和脐血中HBV-DNA载量也越高,且三者的病毒载量依次递减。
3.父亲血液HBV-DNA阳性伴有HBeAg阳性不会增加HBV父婴垂直传播的危险;而父亲血液HBV-DNA阳性伴有精液阳性能增加HBV父婴垂直传播的危险。
4.从分子学水平进一步证实HBV能由父亲传播给后代,父婴基因型一致,以B型为主,C型次之,还分布着少量的D型和B/C混合型。
5.C型的HBV-DNA载量高于其他型。
6.乙肝父婴垂直传播的分布无地域差别。
7.研究提示阻断HBV父婴垂直传播的关键是:孕前定性检测血液或精液HBV-DNA,阳性者,应进行规范化治疗,同时避孕,降至血液或精液中HBV-DNA低水平复制后受孕;建议新生儿出生后24h内和第一个月内注射100U乙肝高效免疫球蛋白。
Objective:
To explore the risk factors of vertical transmission about HBV from HBsAg-positive father to fetal; analyze the relationship of HBV-DNA load and vertical transmission; study whether the genotype is consistent between father and fetal.
Methods:
Using HBsAg and HBV-DNA as screening indicators for pregnant women and their husbands from obstetric clinic from August 2009 to December 2010, totally 154 HBVM were negative or HBsAb was positive only and HBV-DNA was negative pregnant women, HBsAg-positive-husbands and their newborns were enrolled, all subjects provided informed consent for the participation in the study. HBV serological markers were detected by ELISA. HBV-DNA content was detected by fluorescent quantification-PCR (FQ-PCR) and genotypes of HBV were detected by type specific primers and high-resolution melting curve.
Results:
1. Of the 154 neonatal cord blood, 30(19.5%) were HBV-DNA positive. Univariate and multivariate regression analyses were performed to determine risk factors. Maternal HBsAb positive, paternal serum HBeAg-positive, serum HBV-DNA positive, semen HBV-DNA positive, the time of carrying HBV and neonatal sex were statistically significant in the univariate analysis (P<0.05). Multivariate analysis found that HBeAg-positive, serum HBV-DNA positive and semen HBV-DNA positive were still statistically significant (P<0.05).
2. The paternal serum HBV-DNA load was positively correlated with the cord blood HBV-DNA load.
3. The paternal serum HBV-DNA load was positively correlated with pregnant week, but they had a little correlation.
4. Of the 80 semen, 30 (37.5%) were HBV-DNA positive. There was a positive rank correlation between semen HBV-DNA and serum HBV-DNA load levels, the load levels of semen HBV-DNA was lower than that of serum HBV-DNA.
5. The rate of HBsAg positivity for neonatal cord blood is 17.5% (27/154).
6. No relationship was found between serum HBV-DNA load and pregnant week.
7. Among the HBV-DNA load in the paternal serum, semen and cord blood, the paternal serum load was the highest, and the load in cord blood was lowest.
8. The paternal serum HBV-DNA positive was no interaction with HBeAg-positive(P>0.05), on the contrary, paternal serum HBV-DNA positive was interaction with semen HBV-DNA positive (P<0.05).
9. Both the history of pregnancy and neonatal outcome were not statistically significant between the case group and the control group (P>0.05).
10. The HBV-DNA genotype was consistent with father and infant, genotype B is dominant, as well as a small amount of genotype C, genotype D, confection with genotype B and C.
11. Genotype C had a higher serum HBV-DNA level compared with other genotypes.
12. There was no significant difference about paternal HBV vertical transmission in the geographic distribution of Fuzhou.
13. The negative conversion rate was 6.25% when positive infants were followed up to 6 months.
Conclusion:
1. The paternal serum HBV-DNA positive, semen HBV-DNA positive and HBeAg-positive are risk factors of paternal HBV vertical transmission.
2. The HBV-DNA load in umbilical blood are positively correlated with the HBV-DNA load in serum and in semen of father. The load levels of father’s serum HBV-DNA is the highest and the HBV-DNA load in cord blood is the lowest.
3. The serum HBV-DNA positive and HBeAg-positive can not increase the risk, but the serum HBV-DNA positive and the semen HBV-DNA positive can increase the risk about HBV vertical transmission from father to fetal.
4. The study provides further evidence for paternal HBV vertical transmission in molecular level, the genotype is consistent. Genotype B is dominant,as well as a small amount of genotype C, confection with genotype B and C and genotype D.
5. Genotype C has a higher serum HBV-DNA level compared with other genotypes.
6. There is no geographical difference about paternal HBV vertical transmission in urban and suburb in Fuzhou.
7. The research suggest that the key is to test qualitatively HBV-DNA in the serum and semen before pregnancy, positive men should receive effective anti-viral treatment and contraception, until the viral load is low level reproduction. The neonate should be injected 100U Hepatitis B immunoglobulin after the baby is born within 24 hours and the first month respectively.
探讨乙型肝炎病毒(HBV)父婴垂直传播的危险因素;分析血液HBV-DNA载量、精液HBV-DNA载量与父婴垂直传播的关联;研究HBV父婴垂直传播中父婴所携HBV的基因型是否一致。
方法
选择2009年8月至2010年12月在福建省妇幼保健院行产前初诊检查的孕妇,问诊了解其HBsAg且HBV-DNA阴性或仅HBsAb阳性、丈夫HBsAg阳性并决定住本院分娩的孕妇家庭;夫妻双方知情同意参与有关血液或精液乙型肝炎标志物(HBVM)及HBV-DNA定量和基因型检测。分娩期再次抽血确定:孕妇HBVM全阴性或仅HBsAb阳性且HBV-DNA阴性、丈夫HBsAg阳性及其新生儿,共154个研究家庭。采用酶联免疫吸附法(ELISA)检测HBVM,荧光定量PCR法(FQ-PCR)检测血液和精液HBV-DNA载量,型特异性引物和高分辨率溶解曲线测定乙肝病毒基因型。
结果
1.新生儿脐血HBV-DNA阳性率为19.5%(30/154),经单因素分析,孕妇HBsAb阳性、父亲血液HBeAg阳性、父亲血液HBV-DNA阳性、精液HBV-DNA阳性、HBV携带时间、新生儿性别有统计学意义(P<0.05);经多因素分析,父亲血液HBeAg阳性、父亲血液HBV-DNA阳性、精液HBV-DNA阳性仍有统计学意义(P<0.05)。
2.脐血HBV-DNA载量与父亲血液HBV-DNA载量呈正相关。
3.抽血孕周与父亲血液HBV-DNA载量相关性小。
4.精液HBV-DNA阳性率为37.5%(30/80);精液HBV-DNA载量与其血液的HBV-DNA载量呈正相关,精液载量低于血液载量。
5.脐血HBVM检测HBsAg阳性率为17.5%(27/154)。
6.精液HBV-DNA载量与采集孕周相关性不大。
7.HBV-DNA载量由高到低依次为:父亲血液HBV-DNA载量>精液HBV-DNA载量>脐血HBV-DNA载量。
8.父亲血液HBV-DNA阳性伴有HBeAg阳性未见交互作用(P>0.05);父亲血液HBV-DNA阳性伴有精液阳性有交互作用(P<0.05)。
9.两组孕妇孕产史和新生儿妊娠结局等情况类似,均无统计学意义(P均>0.05)。
10.父婴HBV-DNA基因型一致,以B型为主,还有少量的C型、D型和B+C混合型。
11.C型的HBV-DNA载量高于其他型。
12.福州市区和郊县的HBV父婴垂直传播率类似(P>0.05)。
13.阳性新生儿随访至第6个月时,阴转率为6.25%(1/16)。
结论
1.父亲血液HBV-DNA阳性、精液HBV-DNA阳性、血液HBeAg阳性是HBV父婴垂直传播的危险因素。
2.父亲血液HBV-DNA载量越高,其精液和脐血中HBV-DNA载量也越高,且三者的病毒载量依次递减。
3.父亲血液HBV-DNA阳性伴有HBeAg阳性不会增加HBV父婴垂直传播的危险;而父亲血液HBV-DNA阳性伴有精液阳性能增加HBV父婴垂直传播的危险。
4.从分子学水平进一步证实HBV能由父亲传播给后代,父婴基因型一致,以B型为主,C型次之,还分布着少量的D型和B/C混合型。
5.C型的HBV-DNA载量高于其他型。
6.乙肝父婴垂直传播的分布无地域差别。
7.研究提示阻断HBV父婴垂直传播的关键是:孕前定性检测血液或精液HBV-DNA,阳性者,应进行规范化治疗,同时避孕,降至血液或精液中HBV-DNA低水平复制后受孕;建议新生儿出生后24h内和第一个月内注射100U乙肝高效免疫球蛋白。
Objective:
To explore the risk factors of vertical transmission about HBV from HBsAg-positive father to fetal; analyze the relationship of HBV-DNA load and vertical transmission; study whether the genotype is consistent between father and fetal.
Methods:
Using HBsAg and HBV-DNA as screening indicators for pregnant women and their husbands from obstetric clinic from August 2009 to December 2010, totally 154 HBVM were negative or HBsAb was positive only and HBV-DNA was negative pregnant women, HBsAg-positive-husbands and their newborns were enrolled, all subjects provided informed consent for the participation in the study. HBV serological markers were detected by ELISA. HBV-DNA content was detected by fluorescent quantification-PCR (FQ-PCR) and genotypes of HBV were detected by type specific primers and high-resolution melting curve.
Results:
1. Of the 154 neonatal cord blood, 30(19.5%) were HBV-DNA positive. Univariate and multivariate regression analyses were performed to determine risk factors. Maternal HBsAb positive, paternal serum HBeAg-positive, serum HBV-DNA positive, semen HBV-DNA positive, the time of carrying HBV and neonatal sex were statistically significant in the univariate analysis (P<0.05). Multivariate analysis found that HBeAg-positive, serum HBV-DNA positive and semen HBV-DNA positive were still statistically significant (P<0.05).
2. The paternal serum HBV-DNA load was positively correlated with the cord blood HBV-DNA load.
3. The paternal serum HBV-DNA load was positively correlated with pregnant week, but they had a little correlation.
4. Of the 80 semen, 30 (37.5%) were HBV-DNA positive. There was a positive rank correlation between semen HBV-DNA and serum HBV-DNA load levels, the load levels of semen HBV-DNA was lower than that of serum HBV-DNA.
5. The rate of HBsAg positivity for neonatal cord blood is 17.5% (27/154).
6. No relationship was found between serum HBV-DNA load and pregnant week.
7. Among the HBV-DNA load in the paternal serum, semen and cord blood, the paternal serum load was the highest, and the load in cord blood was lowest.
8. The paternal serum HBV-DNA positive was no interaction with HBeAg-positive(P>0.05), on the contrary, paternal serum HBV-DNA positive was interaction with semen HBV-DNA positive (P<0.05).
9. Both the history of pregnancy and neonatal outcome were not statistically significant between the case group and the control group (P>0.05).
10. The HBV-DNA genotype was consistent with father and infant, genotype B is dominant, as well as a small amount of genotype C, genotype D, confection with genotype B and C.
11. Genotype C had a higher serum HBV-DNA level compared with other genotypes.
12. There was no significant difference about paternal HBV vertical transmission in the geographic distribution of Fuzhou.
13. The negative conversion rate was 6.25% when positive infants were followed up to 6 months.
Conclusion:
1. The paternal serum HBV-DNA positive, semen HBV-DNA positive and HBeAg-positive are risk factors of paternal HBV vertical transmission.
2. The HBV-DNA load in umbilical blood are positively correlated with the HBV-DNA load in serum and in semen of father. The load levels of father’s serum HBV-DNA is the highest and the HBV-DNA load in cord blood is the lowest.
3. The serum HBV-DNA positive and HBeAg-positive can not increase the risk, but the serum HBV-DNA positive and the semen HBV-DNA positive can increase the risk about HBV vertical transmission from father to fetal.
4. The study provides further evidence for paternal HBV vertical transmission in molecular level, the genotype is consistent. Genotype B is dominant,as well as a small amount of genotype C, confection with genotype B and C and genotype D.
5. Genotype C has a higher serum HBV-DNA level compared with other genotypes.
6. There is no geographical difference about paternal HBV vertical transmission in urban and suburb in Fuzhou.
7. The research suggest that the key is to test qualitatively HBV-DNA in the serum and semen before pregnancy, positive men should receive effective anti-viral treatment and contraception, until the viral load is low level reproduction. The neonate should be injected 100U Hepatitis B immunoglobulin after the baby is born within 24 hours and the first month respectively.
引文
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[1]杨绍基,任红.传染病学[M].第7版.北京:人民卫生出版社,2008:28.
[2] Lin CL, Kao JH, Chen BF, et al. Application of Hepatitis B Virus Genot ping and Phylogenetic Analysis in Intrafamilial Transmission of Hepatitis B Vir us[J]. Cli Infect Dis 2005; 41:1576–1581.
[3] Komatsu H, Inui A, Sogo T, et al. Source of transmission in children withchronic hepatitis B infection after the implementation of a strategy for prevention in those at high risk[J]. Hepa Res 2009;34:1872-1878.
[4]何进球,袁松华,冼革坚,等.乙型肝炎病毒阳性父-婴垂直传播的临床研究[J].实用妇产科学杂志,2005,21(5):301-302.
[5]周小辉,王莹,温小丽,等.乙型肝炎病毒父婴传播的流行情况研究[J].汕头大学医学院学报,2005,18(4):214-215.
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[9] Ali BA, Huang TH, Salem HH, et al. Expression of hepatitis B virus genein early embryonic cells originated from hamster ova and human spermatozoa transfected with the complete viral genome[J].Asian J Androl,2006;8(3):273–279.
[10]高慧婕,刘超,李倩,等.HBV DNA经小鼠精细胞垂直传递的研究[J].现代医学,2007,35(5):361-364.
[11] Ahmed MM, Huang TH, Xie QD.A Sensitive and Rapid Assay for Investigating Vertical Transmission of Hepatitis B Virus via Male Germ Line Using EGFP Vector as Reporter[J]. J Biome Biote, Volume 2008; Article ID 495436,6 pages.
[12] Wang PL, Wang XH, Cong SY, et al.Mutation analyses of integrated HBVgenome in hepatitis B patients[J]. J Gene Geno2008; 35:85-90.
[13]高慧婕,刘超,李倩.乙型肝炎患者睾丸和附睾组织HBV DNA检测[J].蚌埠医学院学报,2008,33(2):150-152.
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[15]张明辉,王海舫.乙型肝炎病毒父婴垂直传播[J].中国临床医药实用杂志,2004(19):54-56.
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[19]彭桂福,王珊珊,姜普林等.一对父亲与胎儿所携乙型肝炎病毒前S基因的序列分析[J].现代预防医学,2000,27(3):277-279.
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