| 莫艳萍.湖州地区献血人群隐匿性乙型肝炎病毒感染的血清学特征及分子机制研究[J].浙江中西医结合杂志,2021,31(10): |
| 湖州地区献血人群隐匿性乙型肝炎病毒感染的血清学特征及分子机制研究 |
| Serological characteristics and molecular mechanism of occult hepatitis B virus infection in blood donors in Huzhou |
| 投稿时间：2021-03-18 修订日期：2021-04-07 |
| DOI： |
| 中文关键词: 乙型肝炎病毒 隐匿性 无偿献血 基因型 血清学 变异 |
| 英文关键词:Hepatitis B virus Occult Unpaid blood donation Genotype Serology Mutation |
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| 中文摘要: |
| 目的 了解湖州地区献血人群隐匿性乙型肝炎病毒感染(OBI)情况、血清学特点及OBI献血者S区基因突变特征,为OBI感染防控提供参考。方法 获得2018年1月~2019年9月湖州市中心血站无偿献血标本中乙型肝炎表面抗原(HBsAg)初、复检均阴性而核酸扩增检测(NAT)阳性标本作为OBI组,HBsAg初、复检均阳性而NAT阳性标本作为阳性对照组,检测乙肝血清五项,提取血清病毒DNA,实时定光定量聚合酶链式反应法(PCR)测定HBV DNA定量,巢式PCR扩增HBV S基因区,测序,与标准序列比对,观察基因变异情况,发现可能与OBI相关变异位点。结果 2018年1月~2019年9月湖州市中心血站共采集无偿献血标本109 560份,其中HBsAg初、复筛均阴性而NAT阳性标本共检出30份,检出率为0.027%(30/109 560)。OBI血清学模式：抗-HBc(+)/抗-HBs(-)12例(40.00%),抗-HBs(+)/抗-HBc(+)8例(26.67%),抗-HBs(+)/抗-HBc(-)3例(10.00%),抗-HBc(-)/抗-HBs(-)7例(23.33%)；30份OBI标本HBV-DNA病毒载量区间为8~1751(中位数268.33)IU/mL,ALT范围为10~36(中位数18.13)U/L；30份OBI无偿献血样本中29份(96.67%)标本S区扩增成功,2份(6.67%)全基因型扩增成功,23份(76.67%)BCP/PC扩增成功,S区、BCP/PC两个片段均扩增成功22份(73.33%)；基因型分型：B型23例(76.67%),C型5例(16.67%),未明确分型2例(6.67%)；血清型：adw共24例(80.00%),adr共5例(16.67%),ayr共1例(3.33%)；OBI组与阳性对照组HBV基因型均以B基因型占优势,两组基因型分布比较差异无统计学意义(P>0.05)；OBI组小S区18例(62.07%)伴不同程度氨基酸突变,2例(6.90%)OBI提前形成终止密码子；阳性对照组小S区10例(33.33%)出现氨基酸突变,1例(3.33%)提前形成终止密码子,OBI组S区突变率高于阳性对照组(P<0.05)。结论 湖州市献血人员中OBI检出率为0.027%,抗-HBc阳性献血人员OBI感染率较高,基因型以B型为主,HBV S区基因变异可能为OBI发生的关键分子学机制。 |
| 英文摘要: |
| Objective To understand occult hepatitis B virus (HBV) infection (OBI), serological characteristics and gene mutation characteristics of S region in blood donors in Huzhou, so as to provide reference for the prevention of OBI. Methods During the period from January 2018 to September 2019 in central blood station of Huzhou, blood samples with negative hepatitis B surface antigen (HBsAg) in initial examination and re-examination, and with positive HBsAg in nucleic acid amplification test (NAT) were enrolled as OBI group, while blood samples with positive HBsAg in initial examination and re-examination, and with positive HBsAg in NAT were enrolled as positive control group. The five serum makers of hepatitis B were detected. The serum viral DNA was extracted. HBV DNA quantification was detected by real-time quantitative polymerase chain reaction (PCR). HBV S gene region was amplified by nested PCR for sequencing, which was compared with standard sequence. The gene variation was observed to find variation sites that might be related to OBI. Results Amongthe the 109, 560 blood specimens from January 2018 to September 2019 in central blood station of Huzhou, there were 30 cases with negative HBsAg in initial examination and re-examination, and with positive HBsAg in NAT, with detection rate of 0.027% (30/109, 560). In OBI serological mode: 12 cases (40.00%) with anti-HBc(+)/anti-HBs(-), 8 cases (26.67%) with anti-HBs(+)/anti-HBc(+), 3 cases (10.00%) with anti-HBs(+)/Anti-HBc(-), 7 cases (23.33%) with anti-HBc(-)/anti-HBs(-). Of the 30 OBI specimens, interval of HBV-DNA viral load was 8-1751 (median: 268.33) IU/mL and ALT range was within 10-36 (median: 18.13) U/L. Of the 30 OBI blood donation samples, there were 29 (96.67%) cases with successful amplification in S region, 2 (6.67%) cases with successful amplification of whole genotype, 23 (76.67%) cases with successful amplification of BCP/PC, and 22 (73.33%) cases with successful amplification in S region and BCP/PC. In terms of genotype: 23 cases (76.67%) with type B, 5 cases (16.67%) with type C and 2 cases (6.67%) not clearly determined. In terms of serotype: 24 cases (80.00%) with adw, 5 cases (16.67%) with adr and 1 case (3.33%) with ayr. HBV genotypes in both OBI group and positive control group were dominated by type B. There was no significant difference in the distribution of genotypes between the two groups (P>0.05). In OBI group, there were 18 cases (62.07%) in small S area with varying degrees of amino acid mutations and 2 cases (6.90%) with OBI forming stop codons early. In positive control group, there were 10 cases (33.33%) in small S area with varying degrees of amino acid mutations and 1 cases (3.33%) with OBI forming stop codons early. The mutation rate of S region in OBI group was higher than that in positive control group (P<0.05). Conclusion The detection rate of OBI among blood donors in Huzhou is 0.027%. The infection rate of OBI among anti-HBc-positive blood donors is relatively higher. The genotype is mainly on type B. The gene mutation of HBV in S region may be the key molecular mechanism of OBI. |
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