A novel, label-free immunosensor for the detection of α-fetoprotein using functionalised gold nanoparticles

Background and objectives

Novel immunoassay methods based on electrochemical sensors have been developed, but most of these immunosensors are unsuitable for clinical detection because their preparation requires complicated chemical procedures and because their detection sensitivity is restricted. In order to develop a highly sensitive, label-free amperometric sensor for immunoassays, we synthesised novel, functionalised gold nanoparticles (SV-GNP) by covalently capping the surface of gold nanoparticles (GNP) with 1,1′-bis-(2-mercapto)-4,4′-bipyridinium dibromide, a kind of sulfhyrdryl viologen (SV).

Design and methods

We fabricated an immunosensor in a multi-step fashion, by first coating the SV-GNP onto a glassy carbon electrode surface; the resulting electrode core could then adsorb a suitable antibody in a second step to afford the desired immunosensor. α-fetoprotein (AFP) was used as a model analyte in this work.

Results

The anti-AFP/SV-GNP-modified electrode was sensitive to AFP with a linear relationship between 1.25 and 200 ng/mL and a correlation coefficient of 0.9983; the detection limit at a signal to noise ratio of 3 was 0.23 ng/mL under optimal conditions. In addition, the proposed immunosensor exhibited good sensitivity, selectivity, stability and long-term maintenance of bioactivity.

Conclusion

The described immunosensor preparation and immunoassay methods offer promise for label-free, simple, and cost-effective analysis of biological samples.     

A robust electrochemical immunosensor based on core–shell nanostructured silica-coated silver for cancer (carcinoembryonic-antigen-CEA) diagnosis

This work addresses the fabrication of an efficient, novel, and economically viable immunosensing armamentarium that will detect the carcinoembryonic antigen (CEA) typically associated with solid tumors (sarcomas, carcinomas, and lymphomas) and is used as a clinical tumor marker for all these malignancies. We synthesized silver nanoparticles by single-step chemical reduction and coated with silica using a modified Stober method to fabricate silica-coated silver core–shell nanoparticles. The morphologies, structure, and size of the nanoparticles were characterized by Transmission Electron Microscopy (TEM), UV-Visible spectroscopy, X-ray diffraction (XRD), Raman spectroscopy, Fourier Transform Infra-Red Spectroscopy (FTIR), and Dynamic Light Scattering (DLS), respectively. The results indicated that the average size of Ag nanoparticles and silica-coated Ag nanoparticles is 50 nm and 80 nm, respectively. Our TEM results indicate that the silica-shell uniformly encapsulates silver core particles. Further, a disposable electrochemical immunosensor for carcinoembryonic antigen (CEA) was proposed based on the antigen immobilized in a silica-coated silver core–shell nanoparticle film on the surface of an indium–tin–oxide (ITO) flat substrate. The morphological characteristics of the constructed biosensor were observed by scanning electron microscopy (SEM) and electrochemical methods. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were employed for the characterization of the proposed bioelectrode. The cyclic voltammogram appears to be more irreversible on silica coated silver core–shell nanoparticles. It is found that the fabricated immunosensor shows fast potentiometric response under the optimized conditions. The CEA could be determined in the linear range from 0.5 to 10 ng mL⁻¹ with a detection limit of 0.01 ng mL⁻¹ using the interface. The developed flat substrate of ITO for CEA detection (the model reagent) is a potentially promising immunosensing system, manifests good stability, and allows batch fabrication because of its economic feasibility.

The process flow of disposable electrochemical immunosensor fabrication.     

Correlation of improved hepatitis B surface antigen detection limits with hepatitis B virus DNA nucleic acid test yield in blood donations

BACKGROUND: This study provides data on the quantitative relationship between hepatitis B surface antigen (HBsAg; ng/mL) and hepatitis B virus (HBV) nucleic acid test (NAT; copies/mL) and addresses whether HBsAg assays with improved sensitivity would impact the detection of HBV‐positive samples from occult or early seroconversion window period infections. STUDY DESIGN AND METHODS: Plasma samples were tested with an HBsAg assay (PRISM, Abbott Laboratories; sensitivity, 0.08‐0.1 ng/mL) and with two HBsAg research prototype assays: HBsAg Prototype 1 (sensitivity, 0.032‐0.045 ng/mL) and HBsAg Prototype 2 (sensitivity, 0.009‐0.017 ng/mL); NAT assays were used to determine HBV DNA copy levels. RESULTS: Samples from 10 hepatitis B seroconversion panels covering the ramp‐up phase were utilized to examine the relationship between detection of HBsAg using improved assays and viral load using quantitative HBV DNA polymerase chain reaction. For these samples, detection at the HBsAg assay cutoff (sample‐to‐cutoff ratio, 1.0) corresponded to 206 copies/mL HBV DNA for the HBsAg Prototype 1 assay and 329 copies/mL for the PRISM HBsAg assay. Compared to the PRISM HBsAg and HBsAg Prototype 1 assays, the HBsAg Prototype 2 assay detected two additional samples of 32 HBV DNA–positive samples obtained from blood donors with occult HBV and one of seven from blood donors with early window period infections. CONCLUSION: Increased sensitivity HBsAg assays result in the detection of samples containing lower viral loads. Improvements in the analytic sensitivity of HBsAg prototype assays allow the detection of additional HBV DNA–positive samples from donors with window period or occult infections compared to PRISM HBsAg. Improved HBsAg assays should allow for incremental detection of HBV infection.     

乙型肝炎病毒血清标志物GICA检测性能的比对分析

目的评估乙型肝炎病毒血清标志物（HBV—M）胶体金免疫层析法（GICA）的分析性能。方法收集雅培i1000免疫检测系统检测的HBV—M阳性标本653例、阴性标本103例和HBV—M标准品,以化学发光免疫测定法（CLIA）检测结果为标准,分别用酶联免疫吸附试验（ELISA）试剂和5种GICA试剂检测,统计分析检测结果。结果检测HBV—M敏感性：ELISA和GICA检测HBV—M的最低检测限分别为乙型肝炎表面抗原（HBsAg）0．5IU／mL和4～8IU／mL、乙型肝炎表面抗体（抗HBs）10mlU／mL和15～30mlU／mL、乙型肝炎e抗原（HBeAg）1NCU／mL和2～4NCU／mL、乙型肝炎e抗体（抗HBe）2NCU／mL和64～256NCU／mL、乙型肝炎核心抗体（抗HBc）2IU／mL．和32—256IU／mL。检测标本HBV—M的符合率：ELISA分别为87．4％、99．0％、87．7％、96．6％、100．0％,GICA分别为69．6％～71．7％、69．0％～72．0％、70．5％～73．9％、42．2％～49．1％、50．0％～60．0％。检测HBV—M特异性：除GICA柃测抗HBs的特异性略差外,其余各HBV—M的检测特异性均是可接受。,结论GICA检测HBV—M低浓度的标本,漏检率很高,只能用于GICA检测线性范围里HBsAg的筛查,不能作为临床诊断乙型肝炎和疗效考核的依据。     

Prostate-specific antigen detection by using a reusable amperometric immunosensor based on reversible binding and leasing of HRP-anti-PSA from phenylboronic acid modified electrode

BACKGROUND: In recent years, many automated immunoassay analyzers have been developed for accurate diagnosis of various disease states and to improve effective drug administration. Amperometric immunoassay has been increasingly applied to laboratory medicine due to its ease in automation, rapid speed and low detection limits. It is important to develop reusable immunologically-sensitive elements for prostate-specific antigen (PSA) detection. METHODS: The strategy for the immunosensor construction is based on the enzyme-conjugated prostate-specific antibody (HRP-anti-PSA) reversible binding with a self-assembled phenylboronic acid monolayer on gold. RESULTS: After incubating an HRP-anti-PSA modified electrode in a PSA solution, a decrease in the electrocatalytic response of the HRP-anti-PSA modified electrode to the reduction of H(2)O(2) is observed. The photometric activity assays show that this decrease of the electrocatalytic response arises from the formation of immunocomplexes of HRP-conjugated anti-PSA and its antigen, not from the loss of bound HRP-anti-PSA from the electrode surface. Analytical performances and optimal conditions of the described immunosensor are also investigated. Under the optimal conditions, the amperometric immunosensor shows a linear increase of the relative intensity in 2 PSA concentration range from 2 to 15 ng/ml and 15 to 120 ng/ml, respectively. CONCLUSION: This method could be used for rapid analysis of PSA and potentially other antigens.     

Towards cancer diagnostics – an α-feto protein electrochemical immunosensor on a manganese(iv) oxide/gold nanocomposite immobilisation layer

A novel electrochemical immunosensor for the quantification of α-feto protein (AFP) using a nanocomposite of manganese(iv) oxide nanorods (MnO₂NRs) and gold nanoparticles (AuNPs) as the immobilisation layer is presented. The MnO₂NRs was synthesised using a hydrothermal method and AuNPs were electrodeposited on a glassy carbon electrode surface. The MnO₂NRs were characterised with scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM) and X-ray powder diffraction (XRD). Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to characterise the immunosensor at each stage of the biosensor preparation. The MnO₂ nanorods and AuNPs were applied as the immobilisation layer to efficiently capture the antibodies and amplify the electrochemical signal. Under optimised conditions, the fabricated immunosensor was utilised for the quantification of AFP with a wide dynamic range of 0.005 to 500 ng mL⁻¹ and detection limits of 0.00276 ng mL⁻¹ and 0.00172 ng mL⁻¹ (S/N = 3) were obtained from square wave anodic stripping voltammetry and EIS respectively. The nanocomposite modifier enhanced the immunosensor performance. More so, this label-free immunosensor possesses good stability over a period of two weeks when stored at 4 °C and was selective in the presence of some interfering species.

A novel electrochemical immunosensor for the quantification of α-feto protein (AFP) using a nanocomposite of manganese(iv) oxide nanorods (MnO₂NRs) and gold nanoparticles (AuNPs) as the immobilisation layer is presented.     

溶血对乙型肝炎表面抗原检测的影响

目的分析溶血对乙型肝炎表面抗原检测的影响。方法采用金标法检测乙型肝炎表面抗原（HBsAg）。结果溶血标本采用金标法检测HBsAg易产生假阳性或假阴性。结论溶血影响乙型肝炎表面抗原的测定结果 ,溶血标本一般不宜做乙型肝炎表面抗原的检测。     

无标记检测CD4~+T淋巴细胞的免疫传感器构建

目的构建一种无标记检测CD4~+T淋巴细胞的免疫传感器。方法采用葡萄球菌A蛋白(SPA)法定向固定单克隆抗体于金叉指微阵列电极表面以捕获CD4~+T淋巴细胞,并用循环伏安法(CV)扫描对金叉指微阵列电极表面的修饰情况进行表征,最后通过电化学交流阻抗谱法(EIS)对免疫传感器捕获的CD4~+T淋巴细胞进行阻抗检测,通过等效电路拟合获得的阻抗值变化绘制标准曲线。结果该免疫传感器检测CD4~+T淋巴细胞的线性范围是(5.0×103-5.0×106)/mL,检测下限为5.0×102/mL。结论该免疫传感器的结果准确可靠,检测过程简便,价格低廉,有望用于实时检测系统,为实现快速、准确、价廉的CD4~+T淋巴细胞分类计数提供帮助。     

胶体金免疫层析法检测乙型肝炎病毒表面抗原

目的建立一种简易快速、自测式胶体金免疫层析法（ＧＩＣＡ）用于检测血清中乙型肝炎病毒表面抗原（ＨＢｓＡｇ）。方法采用柠檬酸三钠还原法制备胶体金颗粒、标记乙型肝炎病毒表面抗体（抗ＨＢｓ），制成免疫层析检测试条。血清中ＨＢｓＡｇ与测试条上金标记抗体结合后沿着硝酸纤维素膜移动，与膜上的固相抗体结合形成肉眼可见的红色线条。结果ＧＩＣＡ试条只与ＨＢｓＡｇ有特异性反应，与乙型肝炎病毒核心抗原、ｅ抗原等无交叉反应。检测中国药品生物制品检定所ＨＢｓＡｇ标准品，灵敏度可达１ｎｇ／ｍｌ。用ＧＩＣＡ与酶免疫法比较检测了３９５份不同血清标本中ＨＢｓＡｇ，两法符合率为９９．０％。结论ＧＩＣＡ检测血清中的ＨＢｓＡｇ特异性强、灵敏度高、简便快速，无需特殊仪器设备，有广泛应用价值。     

HBV-DNA载量水平与血清学标志物分组模式及前S1抗原的关系

目的:探讨乙型肝炎病毒DNA(hepatitis B virus DNA,HBV-DNA)载量水平与血清学标志物(hepatitis B virus markers,HBVM)分组模式以及前S1抗原(Pre-S1 antigen,Pre-S1Ag)的关系。方法:收集2018年6月至2020年7月来院就诊的180例慢性乙肝患者的血清样本,采用实时荧光定量聚合酶链反应(qRT-PCR)技术检测血清HBV-DNA载量水平;采用化学发光免疫分析法(CLIA)检测乙肝表面抗原(HBsAg)、e抗原(HBeAg)、表面抗体(抗HBs)、e抗体(抗HBe)、核心抗体(抗HBc),并归纳受检样本的HBVM分组模式;采用ELISA法检测血清Pre-S1Ag。分析HBV-DNA载量水平、HBVM分组模式和Pre-S1Ag水平的关系。结果:180例血清样本,HBsAg⁺/抗HBe⁺/抗HBc⁺模式85例(47.22%),HBsAg⁺/HBeAg⁺/抗HBc⁺模式70例(38.89%),其他模式25例(13.89%)。HBsAg⁺/HBeAg⁺/抗HBc⁺模式组HBV-DNA、Pre-S1Ag阳性率均明显高于HBsAg⁺/抗HBe⁺/抗HBc⁺模式组及其他模式组,差异有统计学意义(χ²=56.955、46.809,P<0.05)。将HBV-DNA阳性作为判断HBV复制的金标准,HBeAg的灵敏度为87.23%,特异度为65.12%,阳性预测值为73.21%,阴性预测值为82.35%;Pre-S1Ag的灵敏度为90.35%,特异度为86.36%,阳性预测值为91.96%,阴性预测值为83.82%。依据HBV-DNA载量检测水平,分成<10³ copies/mL、10³~10⁵ copies/mL、10⁵~10⁷ copies/mL、>10⁷copies/mL的4个亚组。随着HBV-DNA载量水平升高,Pre-S1Ag阳性率逐渐升高,分别为41.18%、64.00%、77.78%、94.29%,差异具有统计学意义(χ²=31.250,P<0.05)。结论:HBV血清学标志物和Pre-S1Ag均可用于辅助诊断是否感染HBV以及HBV复制水平,但尚不可取代HBV-DNA定量检测,三者联合检测能为临床诊断、病毒复制提供更全面的依据。     

金标法检测乙肝表面抗原漏检原因分析

目的：分析金标法检测乙肝表面抗原（HBsAg ）的漏检原因。方法采用金标法和酶联免疫吸附法（ELISA）对10335例献血者血液标本进行HBsAg检测,计算阳性率、漏检率。采用金标法和ELISA检测 HBsAg质控品,分析检测灵敏度。结果金标法检测阳性率为0．33％,ELISA为1．14％,金标法检测阳性率低于 ELISA （χ2＝46．76,P＜0．05）。金标法漏检率为71．19％。金标法反应时间为5、10、15 min时,漏检率为94．92％、88．98％、71．19％,比较差异有统计学意义（χ2＝38．27,P＜0．05）。金标法不能检出浓度为0．5 ng/mL的 HBsAg质控品。结论金标法检测灵敏度较ELISA低,并且受人为因素影响较大,适用于献血前初筛。     

时间分辨荧光分析法检测低值乙型肝炎病毒表面抗原的质量评价

目的评价时间分辨荧光分析法检测低值乙型肝炎病毒表面抗原（HBsAg）的性能。方法使用不同浓度试剂盒标准品、二级标准品进行重复测定,建立相应的数学模型,进行检测低限（lower limit of detection,LLD）、生物检测限（biologic limit of detection,BLD）、功能灵敏度（functional sensitivity,FS）、空白限（limit of blank,LoB）、检出限（limit of detection,LoD）、定量检出限（limit of quantifications,LoQ）的统计计算。结果 LLD为0.044ng/mL、BLD为0.18ng/mL、FS为0.18ng/mL、LoB为0.03ng/mL、LoD为0.108ng/mL,尝试计算低值HBsAg检测结果的不确定度,确定定量检出限（LoQ）约为0.5ng/mL。结论使用灵敏度性能和不确定度2种方法进行评价,该检测系统对低值HBsAg分析能够达到试剂盒规定的性能要求。     

滴金法同步检测甲、乙、丙型肝炎病毒免疫标志物方法学建立及评价

目的　建立同步检测甲、乙、丙型肝炎病毒免疫标志物的金免疫渗滤法 (GIFA)的实验方法 ,并对其性能作评价。方法　采用胶体金作为免疫示踪物 ,标记羊抗人IgG单克隆抗体 ,将相应的病毒抗原HAV Ag、HBc Ag及HCV Ag包被在硝酸纤维素薄膜表面 ,标本中所含抗体与相应的抗原结合后 ,加入金标记抗体 ,即可在薄膜表面形成肉眼可见的红色斑点。结果　自制的GIFA纸条法检测 64 5份血清标本 ,与ELISA、RIA法作对比 ,符合率分别为 97.4%、96.9%、10 0 .0 1% ,最低检测浓度分别为 2ng/ml、1ng/ml、1ng/ml。 结论　本实验方法具有较高的特异性和灵敏度 ,操作简便。     

Multiple fluorescent labeling of silica nanoparticles with lanthanide chelates for highly sensitive time-resolved immunofluorometric assays

BACKGROUND: Time-resolved immunofluorometric assays (TrIFA) using lanthanide-labeled nanoparticles have greatly increased the sensitivity of immunoassays. Current labeling strategies, however, use either physical doping of lanthanide chelates into preformed nanoparticles or covalent linking of lanthanide chelates to precursors used for making nanoparticles; both these strategies have drawbacks. METHODS: Luminescent Eu(III) and Tb(III) chelates were covalently coated on the surface of preformed silica nanoparticles to which detection antibodies or bridging proteins for antibody binding were conjugated. We used the resulting conjugates in TrIFA for detection of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg), both individually and simultaneously. We compared the results of the newly established method with results of an ELISA for serum samples. Positive samples identified by TrIFA but not by ELISA were confirmed by additional assays, including real-time PCR detection of viral DNA. RESULTS: The prepared nanoparticle conjugates were homogeneous in size, at approximately 55 (5) nm in diameter [mean (SD)], were stable for long-time storage (>2 years), and contained more chelates [6.86 x 10(5) for Eu(III), 4.73 x 10(4) for Tb(III)] per nanoparticle than particles made as previously reported. The TrIFA established for HBsAg had a comparable or lower detection limit (0.0092 microg/L) than existing nanoparticle-based TrIFA or ELISA. The TrIFA for HBeAg had a much lower detection limit [10.0 National Centre Unit (NCU)/L] than ELISA and detected HBeAg in 5 samples missed by the ELISA method. Simultaneous TrIFA for both HBsAg and HBeAg was achieved with detection limits (0.033 microg/L for HBsAg and 27.0 NCU/L for HBeAg) close to those of the individual assays. CONCLUSIONS: Covalent surface labeling of silica nanoparticles with lanthanide chelates provides good fluorescent labels that can be used in TrIFA for highly sensitive and robust detection of clinical targets.     

全自动酶联免疫检测仪与手工酶联免疫检测的比对评价

目的评价Tecan Freedom Evolyzer-2200全自动酶联免疫仪与手工操作后BIO-RAD680酶标仪检测临床标本结果的一致性。方法用高浓度的乙型肝炎表面抗原（HBsAg）了解仪器的交叉污染情况,采用1IU/mL的HBsAg质控血清检测其孔间重复性和板间重复性,随机抽取50份临床标本经该仪器和手工加样操作BIORAD680酶标仪测定乙型肝炎病毒5项指标,测定结果进行比对。结果经试验该酶联免疫仪器的拖带污染在低水平,与手工操作比对50个项目指标检测结果总的符合率达96.4%。结论该仪器可以满足临床实验需要。     

孕产妇HBV前S1抗原与相关血清标志物联合检测结果分析

目的了解孕产妇乙型肝炎(下称乙肝)病毒(HBV)感染情况,对孕产妇乙肝预防控制提供依据,并探讨HBV前S1抗原和乙肝血清标志物联合检测的关系及临床意义。方法采用酶联免疫吸附试验(ELISA)法对2 067例孕产妇进行血清HBV前S1抗原(PreS1-Ag)、HBsAg、抗-HBs、HBeAg、抗-HBe和抗-HBc进行检测,并对检测结果进行分析。结果 HBsAg阳性211例,阳性率10.21%。在211例HBsAg阳性血清中,PreS1-Ag阳性138例(65.40%),HBeAg阳性71例(33.65%),经χ2检验,二者差异有统计学意义(P<0.05);大三阳模式为HB-sAg(+)、HBeAg(+)、抗-HBc(+),HBsAg(+)、HBeAg(+),小三阳模式为HBsAg(+)、抗-HBe(+)、抗-HBc(+)的PreS1-Ag检测阳性率分别为80.39%、75.00%和64.04%,经χ2检验,3种模式之间差异无统计学意义(P>0.05)。其中71例HBeAg阳性血清标本,PreS1-Ag检出56例(78.87%);HBeAg阴性140例,PreS1-Ag检出82例(58.57%),二者比较差异有统计学意义(P<0.05)。HBsAg阴性血清中未检出PreS1-Ag。结论 PreS1-Ag检测比HBeAg更敏感,是HBV复制的可靠指标之一,与乙肝相关血清标志物联合检测可更准确反映HBV感染与复制状况,及时对孕产妇采取相关措施,对减少母婴传播乙肝,降低新生儿乙肝传播,提高优生优育有着重要意义。     

GP73, a new marker for diagnosing HBV-ACLF in population with chronic HBV infections

Although Golgi protein 73 (GP73) has been widely evaluated for diagnosing hepatocellular carcinoma (HCC) and other liver diseases in recent decade, its serum profile of patients with hepatitis B virus (HBV)–associated acute-on-chronic liver failure (HBV-ACLF) is still unknown. This study was designed to evaluate the serum levels of GP73 in patients with HBV-ACLF. The participants included 200 apparently healthy controls; 200 patients with chronic hepatitis B (CHB); 200 patients with HCC; 210 patients with HBV-ACLF, in which 29 HBV-ACLF patients were followed up for 3 months. All patients were Hepatitis B virus surface antigen (HBsAg) positive. The concentrations of GP73 in patients with HBV-ACLF (285.3 ± 128.5 ng/mL) were markedly higher than those HCC patients (159.1 ± 105.8 ng/mL), CHB patients (64.65 ± 44.99 ng/mL), and healthy controls (35.37 ± 12.41 ng/mL). When the cut-off value was set at 182.1 ng/mL, the sensitivity and specificity of HBV-ACLF diagnosis were 77.62% (95% confidence interval [CI]: 71.37%–83.07%) and 95.50% (95% CI: 92.27%–98.26%), respectively. If serum GP73 concentration was still above 361.6 ng/mL after 14 days of follow-up, the patient's prognosis may be depressed. Serum GP73 may be used to diagnosis HBV-ACLF in population with chronic HBV infections.     

Reagentless and sensitive determination of carcinoembryonic antigen based on a stable Prussian blue modified electrode

Reagentless and sensitive detection of tumor biomarkers using label-free electrochemical immunosensors is highly desirable for early and effective cancer diagnosis. Herein, we present a label-free electrochemical immunoassay platform based on surface-confined Prussian blue (PB) redox probes for sensitive and reagentless determination of carcinoembryonic antigen (CEA). To facilitate the electron transfer of probes and improve sensitivity, Au nanoparticles and PB (Au–PB) are electrochemically co-deposited on a carbon nanotube (CNT) modified glassy carbon electrode (GCE). A polydopamine (pDA) layer is coated on the Au–PB nanocomposite layer in situ as a bifunctional linker. In addition to improving the stability of PB, pDA also provides reducibility for the preparation of gold nanoparticles, which offers an interface for anti-CEA antibody immobilization. The fabricated immunosensor has good stability and is able to reagentlessly detect CEA over a wide range (0.005–50 ng mL⁻¹) with high reproducibility. Furthermore, the immunosensor was used for determination of CEA in human serum samples.

A label-free electrochemical sensor is easily fabricated based on stable and surface-confined Prussian blue for reagentless and sensitive detection of carcinoembryonic antigen.     

乙型肝炎前S1抗原和乙型肝炎血清标志物联合检测结果分析

目的分析乙型肝炎病毒前S1抗原与乙型肝炎病毒(HBV)血清标志物的相互关系。方法用酶联免疫吸附试验对门诊、住院患者血清进行乙型肝炎5项,前S1抗原联合检测(将检测结果中除抗-HBs阳性外的其余项中至少有一项阳性的1591例患者检测结果作回顾性分析)。结果 1591例标本检测结果中,前S1抗原主要出现在HBsAg阳性血清中,前S1抗原在HBeAg阳性组合中的检出率显著高于HBeAg阴性组合,差异有统计学意义(P0.05),在"大三阳"组合中的检出率显著高于"小三阳"组合,差异有统计学意义(P0.05),前S1抗原、HBeAg检出率差异有统计学意义(P0.05)。结论前S1抗原与其他乙型肝炎表面标志物有一定的相关性,在实际工作中两种方法可协同检测,相互补充。     

乙型肝炎病毒表面抗原阴性271例隐匿性感染的研究

目的了解在乙型肝炎表面抗原（HBsAg）阴性人群中隐匿性感染情况。方法采用实时荧光定量聚合酶链反应检测271例重庆黔江区HBsAg阴性住院患者血清中HBV-DNA含量。结果271例HBsAg阴性人群血清HBV—DNA阳性率为10．7％，HBV-DNA含量均低于10^3 copy／mL。HBV—DNA检出率与性别、年龄无关。在HBV-DNA阳性人群中抗-HBc抗体出现率较高，抗-HBs阳性或乙型肝炎病毒标志物全阴性也可检出HBV-DNA。结论血清HBsAg阴性者存在一定比例的隐匿性感染，对不明原因的肝损害、输血、器官移植等应结合灵敏度高的HBV-DNA检测结果再作判断。