Correlation between HBsAg quantitative assay results and HBV DNA levels in chronic HBV

Background

Viral load has been used to diagnose and monitor patients who are being treated for chronic hepatitis B (CHB). The Diagnosis methods are molecular-based and expensive. Quantitation of hepatitis B surface antigen (HBsAg) by automated chemiluminescent micro-particle immunoassay has been proposed to be a surrogate marker. Quantitating HBV DNA levels molecularly is expensive; thus, a cheaper laboratory test as a surrogate diagnostic marker might simplify our management.

Objectives

We determined whether quantitative HBsAg levels correlate with HBV DNA levels in CHB.

Patients and Methods

In this cross-sectional study, all CHB patients who were referred by a gastroenterologist to undergo quantitative HBV DNA assay in a qualified laboratory in Mashhad, Iran in 2009 were enrolled, and blood samples was obtained. Patients who were positive for antibodies to HCV and HDV were excluded. HBV DNA was measured by real-time polymerase chain reaction, and serum HBsAg was quantified byelectrochemiluminescence assay (Roche Diagnostic).

Results

Of 97 patients, 70 were male (72%) and 27 were female (28%); the mean age was 39 ± 11 years. Eighty-seven percent wasHBeAg-negative. By Mann-Whitney test,HBSAg titer differed significantly between HBeAg-positive and -negative patients (P = 0.001), as did HBV DNA levels (P = 0.009). By Spearman test, there was no significant correlation between HBsAg and HBV DNA levels (P= 0.606 and r = 0.53).

Conclusions

HBeAg-negative patients have higher levels of HBsAg and lower levels of HBV DNA. By electrochemiluminescence assay,HBsAg has no significant correlation with HBV DNA levels in CHB with predominant genotype D and HBeAg negativity in Iran.     

Correlation Between Hepatitis B Surface Antigen Titers and HBV DNA Levels

Background/Aim:

To assess the correlation between serum HBsAg titers and hepatitis B virus (HBV) DNA levels in patients with hepatitis B envelop antigen-negative (HBeAg −ve) HBV genotype-D (HBV/D) infection.

Patients and Methods:

A total of 106 treatment- naïve, HBeAg −ve HBV/D patients were included; 78 in the inactive carrier (IC) state and 28 in the active hepatitis (AH) stage. HBV DNA load and HBsAg titers were tested using TaqMan real-time polymerase chain reaction (PCR) and automated chemiluminescent microparticle immunoassay, respectively.

Results:

The median (range) log10 HBsAg titer was significantly lower in the IC group compared with AH group, 3.09 (−1 to –4.4) versus 3.68 (−0.77 to 5.09) IU/mL, respectively; P < 0.001. The suggested cutoff value of HBsAg titer to differentiate between the two groups was 3.79 log10 IU/mL. In addition, there was a significant positive correlation between HBsAg and HBV DNA levels in the whole cohort, AH, and IC groups (r = 0.6, P < 0.0001; r = 0.591, P = 0.001; and r = 0.243, P = 0.032, respectively).

Conclusion:

Serum HBsAg titers may correlate with HBV DNA in treatment-naïve HBeAg –ve HBV/D patients, and supports the use of HBsAg levels in clinical practice as a predictor of serum HBV DNA levels.     

Correlation of Quantitative Assay of HBsAg and HBV DNA Levels During Chronic HBV Treatment

Background and aim Viral load is used for the diagnosis and monitoring the treatment of chronic hepatitis B (CHB). These methods are molecular-based and are expensive. Previous studies suggest that quantitative hepatitis B surface antigen (HBsAg) studied by automated chemiluminescent microparticle immunoassay can be a surrogate marker. In this study, we aimed to investigate whether quantitative HBsAg correlates hepatitis B virus (HBV) DNA levels during CHB treatment. Methods The study included 18 patients (13 male, 5 female, mean age: 33 ± 9 years) with CHB. They were given pegylated interferon ± lamivudine for 52 months and serum samples were obtained in weeks 0, 4, 8, 24, 48, 52, and 76. HBV DNA was measured by TaqMan polymerase chain reaction (PCR; Erasmus MC, University Medical Center, Rotterdam, The Netherlands). Quantitative HBsAg was studied by automated chemiluminescent microparticle immunoassay (Architect HBsAg, Abbott, IL). Results HBV DNA levels were measured as follows: 9.66, 7.69, 7.06, 5.93, 5.89, 5.88, and 7.27 logarithmic genome equivalent/ml, respectively. The corresponding HBsAg quantitation results were 42,888, 31,176, 37,882, 27,277, 28,279, 29,471, and 31,535 IU/ml, respectively. They showed a significant correlation (canonical correlation = 0.85). Conclusions HBsAg studied by automated chemiluminescent microparticle immunoassay correlates with HBV DNA and can be a surrogate marker during the monitoring of the efficacy of HBV treatment.     

HBVDNA的表达与肝纤维化形成的相关性研究

探讨HBVDNA的表达与肝纤维化形成的相关性。应用荧光定量PCR技术测定 2 0 0例乙肝患者和 4 9名健康对照组的血清HBVDNA ,同时用放免法检测层粘连蛋白 (LN)、透明质酸 (HA)、Ⅲ型前胶原 (PⅢ )、Ⅳ型胶原 (Ⅳ .C)水平。HBVDNA表达阳性组LN、HA、PⅢ、Ⅳ .C的含量〔(16 4 8± 5 6 1)、(16 6 1± 78 0 )、(15 3 5± 6 0 7)、(92 1±2 9 9) μg/L〕显著高于HBVDNA阴性组〔(110 9± 2 9 8)、(82 1± 2 4 7)、(91 9± 2 9 2 )、(5 9 5± 14 3) μg/L〕(P <0 0 0 1)和健康对照组〔(10 4 9± 16 3)、(6 8 0± 2 7 6 )、(76 3± 2 1 3)、(5 4 3± 7 8) μg/L〕(P <0 0 0 1)。HBVDNA阳性组中 ,上述四项指标随着HBVDNA含量升高而递增。动态观察 2 8例乙肝患者随着治疗过程HBVDNA浓度下降 ,LN、HA、PⅢ、Ⅳ .C的含量明显递减。HBVDNA的阳性表达与肝纤维化形成的相关性十分密切     

HBsAg阴性/HBV DNA阳性与S基因变异的关系——附8例报告

目的研究无锡地区HBV感染者HBsAg缺失与S基因变异的关系,探讨其形成机理。方法采用ABBOTT测定 HBV M;套式 PCR检测 HBV DNA并作S基因序列分析;荧光定量法测定HBV DNA含量。结果8例HBsAg缺失者S基因出现了变异,导致HBsAg肽63、82、89、90、91、101、115、154位氨基酸替换,且89、90位氨基酸为联合变异;该模式75％(6/8)系慢性肝炎病人,87.5％(7/8)未使用免疫制剂,其HBV DNA含量是低的,而血清抗-HBs中位数值达239.1mU/ml。结论 HBsAg缺失模式多为自然发生的变异,变异主要发生在 a决定簇以外部分,可改变HBsAg的抗原性,使其不能被现行的试剂所检出,此外HBsAg阴性也与血液中HBV DNA含量低有关。     

Detection of HBsAg, HBcAg, and HBV DNA in ovarian tissues from patients with HBV infection

AIM: To investigate the presence of HBsAg, HBcAg, and HBV DNA in ovarian tissues from patients with HBV infection. METHODS: HBsAg and HBcAg were examined in ovarian biopsy tissues from 26 patients with HBV infection by immunocytochemistry, and HBV DNA was detected in ovarian tissues by PCR. RESULTS: HBsAg and HBcAg were present with the same positive rate of 34.6% (9/26). The total positive rate was 46.2% (12/26). HBsAg and HBcAg were positive in 6 (23.1%) of the 26 patients. Brown positive particles were diffusely distributed in ovarian cells. The positive rate of HBV DNA was 58.3% (7/12). CONCLUSION: HBsAg, HBcAg, and HBV DNA can be detected in ovarian tissues from patients with HBV infection. The presence of HBsAg and HBcAg in ovarian tissues does not correlate with the HBV markers in serum.     

乙肝肝硬化血清透明质酸与HBV DNA的相关性

目的:探讨乙肝肝硬化患者血清透明质酸与HBV DNA水平的相关性.方法:收集乙肝肝硬化患者144例,其中女性42例,男性102例,平均年龄(54.42±11.53)岁,Child pugh A级42例,Child pugh B级40例,Child pugh C级62例,采用放射免疫法检测血清透明质酸,荧光定量PCR检测血清HBV DNA水平,对血清透明质酸、血清HBV DNA水平进行统计并分析两者之间的关系.结果:随着乙肝肝硬化Child分级的加重,患者的血清透明质酸水平也增高,不同分级之间的差异有统计学意义(174.10μg/L±127.98μg/Lvs421.35μg/L±176.96μg/Lvs903.58μg/L±212.02μg/L,P<0.01).不同Childpugh分级患者的血清HBV DNA水平差异无统计学意义(P>0.05).不同Child pugh分级患者的血清HBV DNA和透明质酸水平无显著相关性(P>0.05).结论:随着患者的Child pugh分级升高,HA水平显著增高,说明HA是反映肝硬化程度的敏感指标;乙肝肝硬化患者HBV DNA与肝硬化程度无显著相关,血清透明质酸与HBV DNA水平之间也无显著相关性,因此抗病毒和抗纤维化治疗对乙肝肝硬化患者同等重要.     

慢性乙型肝炎患者外周血单个核细胞内HBV RNA、HBV DNA的检测及其与血清HBV DNA的关系

目的:探讨慢性乙型肝炎(慢乙肝)患者外周血单个核细胞(PBMC)中HBV感染情况及复制状态,以及与血清HBV DNA的关系.方法:应用PCR技术特异性,选择地检测慢乙肝患者PBMC中HBV RNA、HBV DNA,并与血清HBV DNA进行比较.结果:在慢乙肝患者PBMC中能检测出HBV RNA及HBV DNA,阳性率分别为38.78%及77.55%,且两者的检出率有一致性.当PBMC中HBV RNA阳性时,HBV DNA均为阳性(100%),而当HBV DNA阳性时,部分病例可表现为HBV RNA阴性,两者阳性符合率为50%.血清中HBV DNA阳性率为71.42%.PBMC中HBV RNA的阳性率与血清中HBV DNA的阳性符合率为63.33%,血清中HBV DNA与PBMC中HBV DNA阳性符合率为76.32%.结果提示:HBV能感染PBMC,在部分患者存在着活动性复制,这种复制表现为不同于肝细胞内的低水平复制.此外,HBV感染PBMC后也存在着非复制状态,表现为PBMC中HBV DNA阳性,但HBV RNA阴性.另外,当血中HBV DNA阴性时,少数病例PBMC仍可表现为HBV RNA及HBV DNA阳性.结论:HBV能够感染PBMC并存在活动性复制,可能是造成患者免疫功能紊乱、肝脏损害慢性化的重要原因之一.     

乙型肝炎孕妇HBV血清标志物与HBVDNA载量及ALT的相关性分析

目的 分析乙型肝炎(乙肝)孕妇HBV血清标志物、HBVDNA载量及ALT检测结果,为HBV感染孕妇的诊治提供参考。方法 回顾性分析2016年11月—2017年11月在我区孕检的120例乙肝孕妇的临床资料,应用酶联免疫吸附法检测血清五项HBV标志物,同时采用荧光实时定量PCR技术检测HBVDNA水平,酶速率法检测ALT,并对检测结果进行统计分析。结果 120例孕妇血清中,感染模式Ⅰ(大三阳)HBsAg(+)、HBeAg(+)、HBcAb(+)58例,占48.33%;HBVDNA(+)49例,占84.48%,其中HBVDNA>106IU/ml42例,占72.41%;ALT增高39例,异常率为67.24%。感染模式Ⅱ(小三阳)HBsAg(+)、HBeAb(+)、HBcAb(+)45例,占37.50%;HBVDNA(+)27例,占60.00%,其中HBVDNA>106IU/ml15例,占33.33%;ALT增高20例,异常率为44.44%。感染模式Ⅰ孕妇HBVDNA阳性率、HBVDNA>106IU/ml率和ALT异常率最高,感染模式Ⅱ孕妇次之。结论 HBV血清标志物与HBVDNA高载量和ALT水平密切相关,三者相结合能为孕妇的临床诊断、围产期干预措施以及疗效观察提供参考依据。     

丁型肝炎患者肝组织HDAg与HBsAg/HBcAg和HBV DNA相关性研究

目的探讨丁型肝炎患者肝组织中HDAg与HBsAg,HBcAg和HBV DNA表达及相关性.方法应用免疫组化双重染色及连续切片技术和原位杂交,检测了79例丁型肝炎患者肝组织HDAg,HBsAg,HBcAg和HBVDNA表达,以52例乙型肝炎作对照.结果丁型肝炎HBsAg,HBcAg检出率为81%,71%,乙型肝炎为94%,92%,两组比较有显著性差异(P＜0.05或0.01).HDAg以肝细胞核表达为主,其次是胞质表达,HBsAg以肝细胞浆表达为主,HDAg和HBsAg表达强度及阳性细胞分布呈一致性,两种抗原的表达程度与肝组织的炎症活动和病理损害相关(P＜0.01).HBcAg以以肝细胞核表达为主,阳性细胞主要呈单个细胞或点状分布,且HBcAg阳性细胞明显少于HDAg阳性细胞.HDAg表达强度与HBV DNA表达呈负相关(P＜0.05).结论HDV感染会抑制HBV DNA复制或病毒抗原表达;在HDV致病机制中HDV的直接细胞毒性可能起主要作用,也有HBV的协同作用.     

HBsAg和HBcAg融合表达质粒对HBV转基因小鼠的体液免疫治疗效应

目的观察融合表达质粒pcDNA3．1-SC对HBV复制和表达的抑制效果。方法 构建融合表达质粒pcDNA3．1-SC，以100μg肌肉接种C57，BL／6 HBV DNA转基因小鼠，2、4周后各加强免疫1次。然后动态检测小鼠血清抗-HBs、抗-HBc的诱生，以及肝组织HBsAg、HBcAg和血清HBV DNA的消长情况。结果接种后4、8、12周，小鼠血清抗-HBs阳转率分别达到55％、67％和33％；而抗-HBc阳转率低于20％；同时，与接种前相比，肝组织内HBsAg、HBcAg表达和血清HBV DNA水平呈逐渐减弱的趋势，在接种后8周，有1／3的小鼠已不能检出。结论融合表达质粒pcD—NA3．1-SC能够在一定程度上抑制转基因小鼠体内的HBV DNA复制和抗原表达，提示了研制乙型肝炎治疗性DNA疫苗的可能性。     

HBV基因型和S基因序列特点与慢性乙型肝炎患者HBsAg/抗-HBs共存关系

目的探讨HBV基因型及S基因序列特点与慢性乙型肝炎（CHB）患者HBsAg和抗-HBs共存的内在相关性以及HBsAg＋/抗-HBs＋的发生机制及意义。方法共收集49例HBsAg＋/抗-HBs＋CHB患者的血清HBVDNA样本,采用PCR-RFLP及测序法鉴定其基因型,并与267例HBsAg＋/抗-HBs-CHB患者的HBV基因型分布进行比较。对6例HBsAg＋/抗-HBs＋CHB患者（Ⅰ组）的HBVS基因进行克隆测序,并与HBsAg＋/抗-HBs-CHB患者（Ⅱ组）进行对比,分析变异种类及频率等的差异。结果 HBsAg＋/抗-HBs＋CHB患者HBV基因型B、C及B/C混合感染的构成比分别为63.3%（31/49）、26.5%（13/49）、10.2%（5/49）,而对照组上述构成比分别为52.8%（141/267）、46.1%（123/267）、1.1%（3/267）,两组构成比差异有统计学意义（P〈0.01）。Ⅰ组HBsAg各区段,特别是主要亲水区（MHR）的变异位点明显多于Ⅱ组;发现了若干新变异、少见的W196和C69终止突变,1例患者检测到G145R变异。某些患者HBsAgMHR-2（包含a决定簇）并未发现变异。结论 HBV基因型的差异及HBsAg变异增多可能是部分CHB患者出现HBsAg＋/抗-HBs＋的原因之一;基因型B或B/C混合感染相对更易出现HBsAg＋/抗-HBs＋。HBsAg变异增多是CHB患者出现HBsAg＋/抗-HBs＋的重要机制之一。某些患者HBsAgMHR-2并无变异,提示HBsAg/抗-HBs共存还存在其他机制。     

Relationship between HBsAg,HBcrAg and hepatocellular carcinoma in patients with undetectable HBV DNA under nucleos(t)ide therapy

We examined the relationship between hepatitis B surface and core‐related antigens (HBsAg, HBcrAg) and hepatocellular carcinoma (HCC) development in patients with undetectable serum HBV DNA receiving nucleos(t)ide analogue (NA). Seventy‐six HBV carriers with undetectable HBV DNA (<20 IU/mL) who subsequently developed HCC were compared with 152 matched controls. Clinical and laboratory parameters (including novel assays to measure linearized HBsAg [HQ‐HBsAg] and HBcrAg) were analysed. There were no significant differences in HBsAg/HQ‐HBsAg levels between the two groups. There was a significant difference in the median values of both pre‐ and post‐NA HBcrAg levels between the HCC and control groups (pre‐treatment: 279.0 vs 35.4 kU/mL, P=.005; post‐treatment: 10.2 vs 1.7 kU/mL, P=.005, respectively). For the whole HCC group, a cut‐off value of post‐treatment HBcrAg level ≥7.8 kU/mL yielded an area under receiver operating curve (AUROC) of 0.61 with a negative predictive value (NPV) of 77.0%. The OR of HCC development was 3.27. For noncirrhotic patients, the median values of post‐treatment HBcrAg level of HCC group and controls were 10.2 and 1.0 kU/mL, respectively (P=.001). A cut‐off value of HBcrAg level ≥7.9 kU/mL yielded an AUROC of 0.70 with a NPV of 80.6%. The OR of HCC development was 5.95. A higher pre‐ and post‐NA treatment HBcrAg level (but not HBsAg) was associated with an increased risk of HCC development in patients achieving undetectable serum HBV DNA while on NA therapy. HBcrAg may serve as a novel risk marker for HCC in this group of patients.     

乙型肝炎患者前S_1抗原与病毒血清学标志物及HBVDNA含量的相关性分析

目的：探讨乙型肝炎患者前S1（Pre-S1）抗原与病毒血清学标志物（HBV-M）、HBV DNA含量的相关性,以评价Pre-S1抗原检测的临床应用价值。方法：检测316例乙型肝炎患者病毒Pre-S1与HBV-M及HBV DNA含量,并进行相关性分析。结果：Pre-S1在HBeAg阳性患者中阳性率为95.7%,与HBeAg及HBV DNA具有相关性（P〈0.01）。结论：Pre-S1与HBV DNA具有相关性,且较HBeAg更敏感,可以作为评价HBV复制的重要指标。