Quantitative hepatitis B core antibody and quantitative hepatitis B surface antigen:Novel viral biomarkers for chronic hepatitis B management

The management of hepatitis B virus(HBV)infection now involves regular and appropriate monitoring of viral activity,disease progression,and treatment response.Traditional HBV infection biomarkers are limited in their ability to predict clinical outcomes or therapeutic effectiveness.Quantitation of HBV core antibodies(qAnti-HBc)is a novel non-invasive biomarker that may help with a variety of diagnostic issues.It was shown to correlate strongly with infection stages,hepatic inflammation and fibrosis,chronic infection exacerbations,and the presence of occult infection.Furthermore,qAnti-HBc levels were shown to be predictive of spontaneous or treatment-induced HBeAg and HBsAg seroclearance,relapse after medication termination,re-infection following liver transplantation,and viral reactivation in the presence of immunosuppression.qAnti-HBc,on the other hand,cannot be relied on as a single diagnostic test to address all problems,and its diagnostic and prognostic potential may be greatly increased when paired with qHBsAg.Commercial qAnti-HBc diagnostic kits are currently not widely available.Because many methodologies are only semi-quantitative,comparing data from various studies and defining universal cut-off values remains difficult.This review focuses on the clinical utility of qAnti-HBc and qHBsAg in chronic hepatitis B management.

Screening and evaluation of human single-chain fragment variable antibody against hepatitis B virus surface antigen

BACKGROUND: Phage display technology has become a vital tool in studies aimed at identifying molecules binding to a specific target. It enables the rapid generation and selection of high affinity, fully human antibody product candidates to essentially any disease target appropriate for antibody therapy. In this study, we prepared the recombinant single-chain fragment variable ( ScFv) antibody to hepatitis B virus surface antigen (HBsAg) by the phage display technology for obtaining a virus-targeting mediator. METHODS: mRNA was isolated from B-lymphocytes from a healthy volunteer and converted into cDNA. The fragment variables of heavy and light chain were amplified separately and assembled into ScFv DNA with a specially constructed DNA linker by polymerase chain reaction. The ScFv DNA was ligated into the phagmid vector pCANT-AB5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13K07 helper phage to form a human recombinant phage antibody library. The volume and recombinant rate of the library were evaluated by bacterial colony count and restriction analysis. After two rounds of panning with HBsAg. the phage clones displaying ScFv of the antibody were selected by enzyme-linked immunosorbant assay ( ELISA) from the enriched phage clones. The antigen binding affinity of the positive clone was detected by competition ELISA. HB2151 E. coli was transfected with the positive phage clone demonstrated by competition ELISA for production of a soluble form of the anti-HBsAg ScFv. ELISA assay was used to detect the antigen binding affinity of the soluble anti-HBsAg ScFv. Finally, the relative molecular mass of soluble anti-HBsAg ScFv was measured by SDS-PAGE. RESULTS: The variable heavy ( VH ) and variable light (VL) and ScFv DNAs were about 340bp, 320bp and 750bp, respectively. The volume of the library was up to 2 × 106 and 8 of 10 random clones were recombinants. Two phage clones could strongly compete with the original HBsAb for binding to HBsAg. Within 2 strong positive phage clones, the soluble anti-HBsAg ScFv from one clone was found to have the binding activity with HBsAg. SDS-PAGE showed that the relative molecular weight of soluble anti-HBsAg ScFv was 32 kDa. CONCLUSION: The anti-HBsAg ScFv successfully produced by phage antibody technology may be useful for broadening the scope of application of the antibody.

Transformation of hepatitis B serologic markers in babies born to hepatitis B surface antigen positive mothers

AIM:To better understand the clinical significance of hepatitis B seroiogic markers in babies born to hepatitis B surface antigen (HBsAg) positive mothers, the incidence of maternal seroiogic markers of hepatitis B via placenta and its transformation in these babies were investigated. METHODS: Mothers with positive HBsAg were selected in the third trimester of pregnancy. Their babies received immunoprophylaxis with hepatitis B immunoglobulin and hepatitis B vaccine after birth, and were consecutively followed up for hepatitis B seroiogic markers and HBV DNA at birth, mo 1, 4, 7, 12, and 24. RESULTS: Forty-two babies entered the study, including 16 born to hepatitis B e antigen (HBeAg)-positive HBsAg carrier mothers and 26 to HBeAg-negative HBsAg carrier mothers. Apart from four babies born to HBeAg-positive carrier mothers and demonstrated persistent positive HBeAg eventually became HBV carriers, all other babies developed anti-HBs before 12 mo of age. Among the other 12 babies born to HBeAg-positive carrier mothers, HBeAg was detected in 7 at birth, in 4 at mo 1, and in none of them thereafter. No antibody response to the transplacental HBeAg was detected. Among the babies born to HBeAg-negative carrier mothers, anti-HBe was detected 100% at birth and mo 1, in 88.5% at mo 4, in 46.2% at mo 7, in 4.2% at mo 12 and none in mo 24. Among all the immunoprophylaxis-protected babies born to either HBeAg-positive or HBeAg-negative carrier mothers, anti-HBc was detected in 100% at birth, mo 1 and mo 4, in 78.9% at mo 7, in 36.1% at mo 12 and in none at mo 24. CONCLUSION: HBeAg can pass through human placenta from mother to fetus and become undetectable before 4 mo of age, but no antibodies response to the transplacental HBeAg can be detected till mo 24 in the immunoprophylaxis-protected babies. The sole existence of anti-HBe before 1 year of age or anti-HBc before 2 years of age in babies born to HBsAg carrier mothers may simply represent the transplacental maternal antibodies, instead of indicators of HBV infection status.

Mutations in surface and polymerase gene of chronic hepatitis B patients with coexisting HBsAg and anti-HBs

AIM： To investigate the clinical significance and presence of mutations in the surface （S） and overlapping polymerase gene of hepatitis B patients with coexisting HBsAg and anti-HBs. METHODS： Twenty-three patients with chronic hepatitis B were studied. Of the 23 patients, i i were both positive for hepatitis B virus （HBV） surface antigen （HBsAg） and antibody to HBV surface antigen （anti-HBs）, 12 were negative for anti-HBs while positive for HBsAg. DNA was extracted from 200 μL serum of the patients. Nucleotide of the surface and overlapping polymerase gene from HBV-infected patients was amplified by PCR, and the PCR products were sequenced. RESULTS： Forty-one mutations were found within the surface gene protein of HBV in 15 patients （10 with coexisting HBsAg and anti-HBs）. Six （14.6%） out of 41 mutations were located at ＂α＂ determinant region in 5 patients （4 positive for HBsAg and anti-HBs）. Eleven mutations （26.8%） occurred in the downstream or upstream of ＂α＂ determinant region. Lamivudine （LMV）- selected mutations were found in three patients who developed anti-HBs, which occurred in amino acid positions （196, 198, 199） of the surface protein and in YMDD motif （M204I/V） of the polymerase protein simultaneously. Presence of these mutations did not relate to changes in ALT and HBV DNA levels.CONCLUSION： Besides mutations in the ＂α＂ determinant region, mutations at downstream or upstream of the ＂α＂ determinant region may contribute to the development of anti-HBs. These mutations do not block the replicating competency of HBV in the presence of high titer of anti-HBs.

HBV C基因型有关的HBsAg阴性HBV DNA阳性患者S区突变对HBsAg的影响

目的通过构建HBV C基因型突变质粒研究HBsAg阴性HBV DNA阳性患者HBV S区突变对HBsAg水平的影响。方法收集2022年8月至2023年4月首都医科大学附属北京佑安医院107例HBsAg-/HBV DNA+患者血液样本,对成功提取扩增的HBV DNA S区进行测序,通过构建HBV C基因型突变质粒对HBV S区突变位点进行细胞功能验证,探讨OBI可能发生的分子机制。结果对成功提取扩增的68例患者进行测序,发现HBV S区存在大量突变,包括免疫逃逸突变(如sG145R、sK122R、sS114T、sT131P等)和跨膜结构域(transmembrane domain,TMD)突变(如sT5A、sG10D、sF20S等)。通过构建HBV C基因型突变质粒,进行细胞转染和细胞免疫荧光实验发现sG145R突变会明显降低HBsAg的表达,但是sK122R、sI26N、sQ29N、sR169H、sS114T、sT131P这6个突变位点并未影响细胞内外HBsAg的表达。结论通过测序发现HBsAg-/HBV DNA+患者HBV S区存在大量突变位点,通过构建sG145R、sK122R、sI26N、sQ29N、sS114T和ST131P等突变质粒发现sG145R突变会明显降低细胞内外HBsAg的表达,但是sK122R、sI26N、sQ29N、sR169H、sS114T、sT131P并未明显降低细胞内外HBsAg的表达。

Differential reactivity of mouse monoclonal anti-HBs antibodies with recombinant mutant HBs antigens

AIM: To investigate the reactivity of a panel of 8 mouse anti-hepatitis B surface antigen (HBsAg) monoclonal antibodies (mAbs) using a collection of 9 recombinant HBsAg mutants with a variety of amino acid substitutions mostly located within the “a” region.METHODS: The entire HBs genes previously cloned into a mammalian expression vector were transiently transfected into COS7 cells. Two standard unmutated sequences of the ayw and adw subtypes served as controls. Secreted mutant proteins were collected and measured by three commercial diagnostic immunoassays to assess transfection efficiency. Reactivity of anti-HBs mAbs with mutated HBsAgs was determined by sandwich enzyme-linked immunosorbent assay (ELISA).RESULTS: Reactivity of anti-HBs mAbs with mutated HBsAgs revealed different patterns. While three mutants reacted strongly with all mAbs, two mutants reacted weakly with only two mAbs and the remaining proteins displayed variable degrees of reactivity towards different mAbs. Accordingly, four groups of mAbs with different but overlapping reactivity patterns could be envisaged. One group consisting of two mAbs (37C5-S7 and 35C6-S11) was found to recognize stable linear epitopes conserved in all mutants. Mutations outside the “a” determinant at positions 120 (P→S), 123(T→N) and 161(M→T) were found to affect reactivity of these mAbs.CONCLUSION: Our findings could have important implications for biophysical studies, vaccination strategies and immunotherapy of hepatitis B virus (HBV) mutants.

Is there a need for universal double reflex testing of HBsAg-positive individuals for hepatitis D infection?

Hepatitis D virus(HDV)can infect HBsAg-positive individuals,causing rapid fibrosis progression,early decompensation,increased hepatocellular carcinoma risk,and higher mortality than hepatitis B virus(HBV)mono-infection.Most countries lack high-quality HDV prevalence data,and the collection techniques employed often bias published data.In recent meta-analyses,HDV prevalence in HBsAg-positive patients reaches 5%-15%and is even significantly higher in endemic areas.Since HBV vaccination programs were implemented,HDV prevalence has decreased among younger populations.However,owing to immigrant influx,it has increased in some Western countries.The current practice of HDV screening in HBsAg-positive individuals is stepwise,based on physician’s discretion,and limited to at-risk populations and may require numerous visits.Double reflex testing,which includes anti-HDV testing in all HBsAg-positive individuals and then HDV RNA testing for anti-HDV-positive ones,is uncommon.Reflex testing can identify more HDV infection cases and link identified patients to further care and follow-up.Moreover,laboratory-based double reflex screening is less biased than physician-led testing.Therefore,health-care providers should learn about reflex testing,and federal and provincial hepatitis control programs should implement laboratory-based double reflex testing to obtain reliable HDV prevalence estimates.The test’s cost-effectiveness depends on the number of HBV-positive patients screened to identify one HDV-positive patient.Such testing may be viable in areas with low HBsAg but high HDV prevalence.However,its economic impact on areas with low HDV prevalence needs further study.

8种国产HBsAg试剂盒检测变异HBsAg的效果评价

目的评价国内8种HBsAg诊断试剂检测体外表达的18种变异HBsAg的效果。方法用8种ELISA试剂检测真核细胞表达的18种变异HBsAg的免疫反应性。结果8种试剂盒检测变异HBsAg的能力不同,漏检率为16.7%～44.4%。K122I、T123N、C124R和G145R变异的HBsAg漏检率最高。结论8种HBsAg检测试剂盒都不能很好地检出部分变异HB-sAg,应改进试剂性能,提高对变异HBsAg的检出率。

HBsAg Mab胶体金探针制备与鉴定的实验研究

目的:研制合格的乙肝表面抗原单克隆抗体(HBsAg Mab)标记的胶体金探针。方法:采用柠檬酸三钠还原法制备15nm的胶体金;所制备的胶体金通过透射电镜和紫外分光计度计鉴定其大小和均匀度。通过CVAI曲线,确定胶体金标记HBsAg Mab蛋白用量;采用斑点免疫吸附试验对探针进行鉴定。结果:制备的15nm胶体金颗粒均匀;紫外分光光度计400-700nm扫描结果最大吸收波长为518nm,峰宽较窄;纯化的HBsAg Mab浓度为65mg/mL;在PH为8．2时,每毫升胶体金的蛋白最适保护量为32．5μg;采用斑点免疫吸附试验鉴定探针质量合格,可保存3个月。结论:制备的HBsAg Mab胶体金探针质量合格,为进一步研究提供手段。

HBsAg与抗-HBs同时阳性的慢性乙型肝炎患者临床特点分析

目的分析HBsAg与抗-HBs同时阳性的现象及其临床特点,并探讨其产生的原因。方法收集2011年2月-2014年2月东南大学附属第二医院体检者2260例,其中被诊断为慢性乙型肝炎的患者830例。采用化学发光微粒子免疫分析法筛选HBsAg与抗-HBs同时阳性的患者188例,分为HBeAg阳性组(n=101)和HBeAg阴性组(n=87)。同时选取200例HBsAg阳性、抗-HBs阴性者作为对照,其中HBeAg阳性组80例,HBeAg阴性组120例。检测HBV血清学标志物、肝功能、病毒载量并结合临床进行分析。计数资料组间比较采用χ2检验。结果 HBV血清学标志物在HBsAg与抗-HBs双阳性情况下共有5种模式,其中以HBsAg、抗-HBs、HBeAg及抗-HBc阳性,且抗-HBe阴性多见,占47.9%(90/188),肝功能指标总异常率为69.1%(130/188),HBV DNA总阳性率为56.9%(107/188)。HBeAg阳性的2组HBV DNA均存在高水平复制,其中HBsAg与抗-HBs双阳性组HBV DNA阳性率与对照组比较,差异无统计学意义(χ2=2.632,P>0.05);HBeAg阴性组中,HBsAg与抗-HBs双阳性组HBV DNA定量>1×105IU/ml的比例与对照组比较,差异有统计学意义(χ2=10.740,P<0.05)。对HBV S区进行测序分析发现,测序的80例HBsAg与抗-HBs双阳性患者中有27例患者的HBV S区发生变异,突变率33.7%,且S区变异位点主要有P29L、S61L、P62L、I126T/S、Q129N、M133K、F134L、G145R/K、L175S和L186H等。结论 HBsAg与抗-HBs同时阳性者在乙型肝炎患者中有一定比例,其主要原因可能是病毒株变异所致。这种情况并不代表疾病好转,且抗-HBs出现并不一定能完全有效清除HBsAg,病毒DNA往往存在持续复制,需引起重视。

乙肝患者血清HBsAg与HBsAb双阳性的临床分析

目的通过分析乙型病毒性肝炎(以下简称"乙肝")患者乙型肝炎病毒(HBV)血清标志物中乙型肝炎表面抗原(HBsAg)与乙型肝炎表面抗体(HBsAb)同时阳性罕见模式的病例,探讨其存在原因及临床价值。方法应用微粒子酶免疫化学发光分析法(MEIA)检测乙肝患者血清标志物(HBV-M);采用用荧光定量PCR方法检测HBV-DNA,采用PCR-高分辨率熔解曲线(PCR-HRM)分析技术检测HBV基因基本核心启动子(BCP)双变异nt1762A-T/nt1764G-A;比色法检测谷丙转氨酶和谷草转氨酶,并结合患者的临床特点进行综合分析。结果在15 600例乙肝患者中,HBsAg与HBsAb双阳性的检出率为2.3%,其阳性率在不同性别组及不同年龄组中差异无统计学意义(P> 0.05);在所有HBsAg与HBsAb双阳模式中,HBsAg、HBsAb、HBeAb、HB-cAb阳性模式比例最高,占57.9%;HBsAg与HBsAb双阳模式中HBeAg阳性组乙肝患者的HBV DNA阳性率高于HBeAg阴性组;而HBeAg阴性组BCP双变异nt1762A-T/nt1764G-A发生率较HBeAg阳性组高;两组差异具有统计学意义(P <0.05);慢性乙肝患者HBsAg与HBsAb双阳检出率均高于无症状携带者、乙肝后肝硬化、重型乙肝及乙肝肝癌(P <0.05)。结论 HBsAg与HBsAb双阳性出现,并不预示乙肝病毒传染性减弱,而提示病毒基因有可能出现变异,临床有必要通过检测血清HBV DNA来确定患者体内病毒是否处于复制状态,为此部分HBsAg与HBsAb双阳性患者的个性化治疗提供一定临床帮助。

医院就诊人群HBsAg与抗HBs双阳性HBV感染者的流行病学与分子病毒学特征

目的探讨乙型肝炎病毒表面抗原(HBsAg)和抗HBsAg抗体(抗HBs)同时阳性的HBV感染者的流行病学及分子病毒学特征。方法分析52 070例医院就诊人群HBV血清学标志物检测结果,以HBsAg和抗HBs双阳性患者为实验组,HBsAg阳性、抗HBs阴性患者为对照组,比较2组患者的流行病学特征。半巢氏PCR扩增2组患者HBV S蛋白编码区并测序,比较2组间在不同基因区、基因型及临床诊断时的统计学差异。结果医院检测人群HBsAg阳性率为20.40%(10 621/52 070),其中HBsAg、抗HBs双阳性率为2.48%(263/10 621)。HBsAg、抗HBs双阳性在0~9岁和≥80岁人群流行率较高,而HBsAg阳性、抗HBs阴性则相反。实验组S蛋白氨基酸突变率显著高于对照组(1.52%vs 0.81%,P<0.01),差异主要存在于主要亲水区(MHR)(1.68%vs 0.57%,P<0.01)。实验组中除肝癌(HCC)患者外(1.97%vs 2.21%,P>0.05),乙肝携带者、慢性乙型肝炎(CHB)患者和肝硬化(LC)患者的S蛋白突变率均显著高于对照组同疾病患者(分别为1.47%vs 0.65%,1.28%vs0.84%,2.21%vs 0.44%,P均<0.05)。结论 HBsAg、抗HBs双阳性以0~9岁和≥80岁HBsAg阳性人群更多见。HBV感染者血清HBsAg和抗HBs共存可能与S蛋白(主要为MHR)的突变造成的免疫逃逸有关。HBsAg、抗HBs双阳性和HBsAg阳性、抗HBs阴性患者之间S蛋白的突变率差异与患者肝脏疾病所处的阶段有关。

HBV感染者血清HBsAg、抗-HBs共存与S基因变异

目的 :探讨无锡地区HBV感染者HBsAg与抗 -HBs的共存模式与S基因变异、病毒复制及免疫治疗的关系。方法 :采用Abbott及ELISA测定HBVM ;套式PCR检测HBVDNA并作序列分析 ;荧光定量法检测HBVDNA含量。结果 :30例adr亚型 ,1例adw亚型 ;2 1份血清出现S区多部位变异 ,造成HBsAg肽 36、47、6 3、77、89、90、115、12 6、12 9、139、15 4位氨基酸替换 ;该变异不影响病毒复制 ;变异组免疫药物的使用率 6 6 .6 % ;抗 -HBs阳性血清加入HBsAg阳性血清后 ,再测抗 -HBs阴转。结论 :S基因变异导致HBsAg抗原性的改变和与抗 -HBs结合力的降低 ,以及检测灵敏度的提高、亚型的转变都是造成HBsAg与抗 -HBs共存的原因 ;

Elecsys HBsAg确证试验在灰区及弱反应性标本检测中的应用

目的探讨ElecsysHBsAg确证试验在ELISA定性试验和ECLIA定量试验临界灰区及弱反应性标本检测中的临床应用价值。方法所有被检测血清标本均来自门诊和住院患者，对其中145例ELISA法检出的灰区及弱反应性标本分别进行E1ecsysHBsAg定量试验及确证试验，并对结果进行了比较分析。结果血清HBsAg经EUSA法检测OD／CUTOFF结果在0．85～O．99之间标本共35例，其中1例ElecsysHBsAg定量及确证试验阳性占2．9％（1／35）；OD／CUTOFF结果在1．0～1．99弱反应性标本共54例，ElecsysHBsAg定量试验有反应性14例，占25．9％（14／54），ElecsysHBsAg确证试验阳性18例占33．3％（18／54）；OD／CUTOFF结果在2．0～4．99共31例，有17例ElecsysHBsAg定量及确证试验阳性占54．8％（17／31）；OD／CUTOFF结果在5．O～7．99共25例，有21例ElecsysHBsAg定量及确证试验阳性占84．0％（21／25）。通过对EuSA法与ECLIA法、ELISA法与ElecsysHBsAg确证试验检测HBsAg结果比较差异具有统计学意义（P〈O．01）。结论ELISA法检测HBsAg灰区及较低含量的弱反应性标本、可疑或乙肝感染模式不常见的标本需再次用敏感性更高的ECLIA等方法复检以防假阴性或假阳性出现，有条件的实验室可用Elect；ysHBsAg确证试验进一步确认。

输血前患者血HBsAg、抗HCV、抗HIV和梅毒抗体检测结果临床研究

目的:观察输血前患者血乙肝表面抗原(HBsAg)、丙肝抗体(抗HCV)、艾滋病抗体(抗HIV)和梅毒抗体检测结果。方法:取医院输血治疗患者3542例,治疗前对患者采用化学发光免疫测定(CLIA)法和ELISA法检测血清标本中血HBsAg、抗HCV、抗HIV和梅毒抗体指标,分析其检测结果并提出相应的解决对策。结果:3542例输血患者中704例血HBsAg阳性,52例抗HCV阳性、3例抗HIV阳性,200例梅毒抗体阳性;男性血HBsAg、抗HCV、抗HIV和梅毒抗体感染率,高于女性(P<0.05)。3542例输血患者中不同年龄段均存在血HBsAg、抗HCV、抗HIV和梅毒抗体感,对于血HBsAg、抗HCV多发生在21~40岁年龄段;抗HIV多发生在41~60岁年龄段;梅毒抗体发生在41~>60岁年龄段。959例血HBsAg、抗HCV、抗HIV和梅毒抗体感染病例分布科室较多,排在前三位分别为消化内科、普外科及肿瘤科,分别占33.98%、20.96%及17.00%。结论:输血前加强患者血HBsAg、抗HCV、抗HIV和梅毒抗体指标检测,能降低血液传染性疾病发生率,值得推广应用。

弓形虫和乙型肝炎病毒混合核酸疫苗的研究Ⅲ.pcDNA3-HBsAg-GRA1免疫小鼠的保护性观察

目的观察重组质粒pcDNA3HBsAgGRA1DNA接种诱导的保护性免疫应答。方法质粒DNA免疫BALB/c小鼠;ELISA法检测GRA1、HBsAg抗体及亚类水平;提取各免疫组小鼠肌肉组织DNA进行PCR检测;弓形虫RH强毒株攻击感染各免疫组小鼠。结果经pcDNA3HBsAgGRA1免疫组小鼠产生抗GRA1和HBsAg抗体,且抗GRA1的抗体水平明显高于GRA1单独和GRA1与HBsAg混合免疫组。弓形虫RH强毒株攻击感染pcDNA3HBsAgGRA1免疫组小鼠,其存活时间明显长于其他各组,结果提示HBsAg可能起免疫佐剂作用。结论将GRA1与HBsAg融合明显增强了GRA1的免疫原性和保护性。

非竞争性ELISA法测定人源抗HBsAg Fab功能性亲和常数

目的 测定完全人源化基因工程抗体HBsAgFab的亲和常数。方法 采用非竞争性ELISA固相法 ,经确定最佳抗原包板浓度、最佳抗原包板时间及最佳抗原与抗体结合反应时间后 ,得到了HBsAg与抗体片段抗HBsAgFab及完整抗体抗HBsAgIgG的抗原抗体结合反应曲线 ,计算出抗HBsAgFab及抗HBsAgIgG的亲和常数。结果 人源基因工程抗体抗HBsAgFab的功能性亲和常数在 10 7～10 8M 1水平 ,比完整抗HBsAgIgG仅仅小约 1个数量级 (10 8～ 10 9M 1)。结论 该基因工程抗体与抗原结合能力较强 ,为今后开发应用Fab进行生物导向治疗提供了理论基础。

重庆市2008～2012年无偿献血者HBsAg,ALT及抗HIV、抗HCV、抗TP抗体检测结果的分析

目的分析重庆市2008~2012年无偿献血者乙型肝炎病毒表面抗原（HBsAg）,丙氨酸氨基转移酶（ALT）及抗丙型肝炎病毒（HCV）、抗人类免疫缺陷病毒（HIV）、抗梅毒螺旋体（TP）抗体检测结果,为招募低危无偿献血者,减少血液报废提供依据。方法选择2008~2012年于重庆市血液中心接受酶联免疫吸附测定（ELISA）筛查的无偿献血者的血液标本551 133例,检测其HBsAg,ALT及抗HIV、抗HCV、抗TP抗体。采用实验室信息管理系统ITSWELL对检测结果进行读取、保存、汇总,并判断标本是否合格。结果共计检出不合格标本37 534例（6.81%）,其中,HBsAg,ALT及抗HCV、HIV、TP抗体不合格率分别为1.10%、3.79%、0.51%、0.33%、1.08%。结论加强血液检测试剂筛选及无偿献血者队伍的建设将有助于临床的输血安全。

乙型肝炎表面抗原(HBsAg)胶体金试剂的研制

目的 研制乙型肝炎表面抗原 (HBsAg)胶体金试剂。方法 用未加氢氧化铝佐剂的乙型肝炎血源疫苗原材料免疫BALB/c小鼠制备抗 -HBs单克隆抗体 ;应用胶体金标记抗 -HBs单克隆抗体 ;测定HBsAg胶体金试剂的稳定性 ;对该试剂进行临床考核及检定等。结果 经中国药品生物制品检定所检定 ,其灵敏度达到 1ng/ml,特异度为 10 0 % (2 0 / 2 0 ) ,精密度为 10 0 % (10 / 10 ) ,符合国家对HBsAg试剂盒的要求 ,临床考核合格 ,已通过国家药品监督管理局的新药评审 ,获国家新药证书。结论 HBsAg胶体金试剂可用于献血员HBsAg筛查、健康人群体检、乙型肝炎病毒 (HBV)感染的流行病学调查和临床诊断。