A novel bead-based fluorescence immunoassay for aldosterone

Aldosterone quantification helps evaluate the rennin-angiotensin-aldosterone system. The new bead-based mul-tiplex platform has not been applied in aldosterone detection to achieve simultaneous measurements of multiple hormones. A new sensitive competitive bead immunoassay based on Luminex technology for detecting aldoster-one in small sample volumes was developed using two-antibody coupled beads and biotinylated aldosterone as tracer in combination with an extraction step. The assay was validated in human and mouse samples and exhibited a linear working range from 10 to 1,000 pg/mL. The assay was reproducible and precise with intra-assay coeffi-cient of variations （CVs） from 6.0% to 11.2%, inter-assay CVs from 8.0% to 13.0% and good recovery [（90-110）%] and linearity [（89-107）%]. Excellent correlation was found between this new assay and the reference method （r = 0.96, P 0.000,1）. The successful establishment of this assay provides high possibility for carrying out bead-based multiplex assay measuring aldosterone and other parameters simultaneously in one 50 μL sample so that the efficiency can be improved and precious samples can be saved.

Bead-based microfluidic platforms for multiplex and ultrasensitive immunoassays in clinical diagnosis and treatment

Multiplex ultrasensitive detection of low abundance proteins remains a significant challenge in clinical applications,necessitating the development of innovative solutions.The integration of bead-based microfluidic chip platforms with their efficient target capture and separation capabilities,along with the advantages of miniaturization and low reagent consumption,holds great promise for building an integrated point-of-care testing(POCT)system that enables seamless sample input-result output.This review presents a comprehensive overview of recent advancements in bead-based microfluidic platforms for multiplex and ultrasensitive immunoassays,along with their potential applications in clinical diagnosis and treatment,which is organized into four sections:encoding techniques,the role of microfluidic platforms,applications,and future prospects.

Methodical and pre-analytical characteristics of a multiplex cancer biomarker immunoassay

AIM: To test the methodical and pre-analytical performance of a new multiplex cancer biomarker panel using magnetic beads. METHODS: The MILLIPLEX? MAP Human Circulating Cancer Biomarker Magnetic Bead Panel 1 comprises the tumor markers carcinoembryonic antigen, alpha-fetoprotein, total prostate-specific antigen, cancer antigen 15-3, cancer antigen 19-9, cancer antigen 125, cytokeratine 19-fragment, β-human chorionic gonadotropin, human epididymis protein 4, osteopontin, prolactin, the cell death and angiogenesis markers soluble Fas, soluble Fas-ligand, tumor necrosis factor related apoptosisinducing ligand, vascular endothelial growth factor andthe immunological markers interleukin-6(IL-6), IL-8, tumor necrosis factor-α, transforming growth factor α, fibroblast growth factor-2, macrophage migration inhibitory factor, leptin, hepatocyte growth factor, and stem cell factor. We determined intra- and inter-assay imprecision as well as dilution linearity using quality controls and serum pools. Furthermore, the stability of the 24 biomarkers examined in this panel was ascertained by testing the influence of different storage temperatures and time span before centrifugation.RESULTS: For all markers measured in the synthetic internal quality controls, the intra-assay imprecision ranged between 2.26% and 9.41%, while for 20 of 24 measured markers in the physiological serum pools, it ranged between 1.68% and 12.87%. The inter-assay imprecision ranged between 1.48%-17.12% for 23 biomarkers in synthetic, and between 4.59%-23.88% for 18 biomarkers in physiological quality controls. Here, single markers with very low concentration levels had increased imprecision rates. Dilution linearity was acceptable(70%-130% recovery) for 20 biomarkers. Regarding pre-analytical influencing factors, most markers were stable if blood centrifugation was delayed or if serum was stored for up to 24 h at 4 ℃ and 25 ℃ after centrifugation. Comparable results were obtained in serum and plasma for most markers. However, great changes were observed for single markers.CONCLUSION: MILLIPLEX? MAP Human Circulating Cancer Biomarker Magnetic Bead Panel 1 assay is a stable and precise method for detection of most biomarkers included in the kit. However, single markers have to be interpreted with care.

Precision medicine allergy immunoassay methods for assessing immunoglobulin E sensitization to aeroallergen molecules

Molecular-based allergy diagnosis for the in vitro assessment of a patient immunoglobulin E(IgE) sensitization profile at the molecular level uses allergen molecules(also referred to as allergen components), which may be well-defined, highly purified, natural allergen components or recombinant allergens. Modern immunoassay methods used for the detection of specific Ig E against aeroallergen components are either singleplex(such as the fluorescence enzyme immunoassay with capsulated cellulose polymer solid-phase coupled allergens, the enzyme-enhanced chemiluminescence immunoassay and the reversed enzyme allergosorbent test, with liquid-phase allergens), multiparameter(such as the line blot immunoassay for defined partial allergen diagnostics with allergen components coating membrane strips) or multiplex(such as the microarraybased immunoassay on immuno solid-phase allergen chip, and the two new multiplex nanotechnology-based immunoassays: the patient-friendly allergen nanobead array, and the macroarray nanotechnology-based immunoassay used as a molecular allergy explorer). The precision medicine diagnostic work-up may be organized as an integrated "U-shape" approach, with a "top-down" approach(from symptoms to molecules) and a "bottomup" approach(from molecules to clinical implications), as needed in selected patients. The comprehensive and accurate Ig E sensitization molecular profiling, with identification of the relevant allergens, is indicated within the framework of a detailed patient's clinical history to distinguish genuine Ig E sensitization from sensitization due to cross-reactivity(especially in polysensitized patients), to assess unclear symptoms and unsatisfactory response to treatment, to reveal unexpected sensitizations, and to improve assessment of severity and risk aspects in some patients. Practical approaches, such as anamnesis molecular thinking, laboratory molecular thinking and postmolecular anamnesis, are sometimes applied. The component-resolved diagnosis of the specific IgE repertoire has a key impact on optimal decisions making for prophylactic and specific immunotherapeutic strategies tailored for the individual patient.

多重细胞因子检测试剂的开发及其在骨髓瘤中的应用

目的:开发多重细胞因子检测试剂,分析复发难治性多发性骨髓瘤(RRMM)的细胞因子、血管生成、骨重塑蛋白表达水平变化。方法:采用多重微球流式荧光免疫技术建立多重细胞因子定量检测试剂,并应用于55例RRMM患者和22例健康对照血清样本的测定,分析患者细胞因子、血管生成、骨重塑蛋白表达水平及其与临床特征的相关性。结果:本研究成功开发了十重细胞因子免疫分析检测试剂,平均灵敏度为7.1 pg/ml,平均回收率为97.4%,平均批内变异系数为4.8%,平均批间变异系数为9.0%。此外,RRMM样本结果发现IL-2、IL-17、DKK1、RANKL、OPG水平与IgG单克隆蛋白水平呈正相关,TIMP1与IgG、IgA单克隆蛋白水平呈正相关。结论:本研究开发出10种具有较高灵敏度和特异性的细胞因子检测试剂,并发现IL-2、IL-17、DKK1、RANKL、OPG、TIMP1在RRMM中具有追踪疾病进程的潜在价值。建立的多重细胞因子试剂开发流程对今后拓展蛋白标志物多重检测试剂的研发和应用具有重要参考价值。

A microfluidic biosensor for multiplex immunoassay of foodborne pathogens agitated by programmed audio signals

Foods are often contaminated by multiple foodborne pathogens,which threatens human health.In this work,we developed a microfluidic biosensor for multiplex immunoassay of foodborne bacteria with agitation driven by programmed audio signals.This agitation,powered by the vibration of a speaker cone during music playing,accelerated the mass transport in the incubation process to form bacterial complexes within 10 min.Immunoassay reagents of the two target bacteria(Escherichia coli O157:H7 and Salmonella typhimurium)were preloaded into the corresponding fore-vacuum storage chamber on the chip,and released to participate in the subsequent immune analysis process by piercing the chambers.All the detection processes were integrated into a single microfluidic chip and controlled by a smartphone through Bluetooth.Under selected conditions,wide linear ranges and low limits of detection(LODs<2CFU/m L)were obtained,and real food samples were successfully determined within 30 min.This biosensing method can be extended to wide-ranging applications by loading different recognizing reagents.

化学发光免疫分析方法与应用进展

化学发光免疫分析方法具有选择性好、灵敏度高、分析速度快、设备简单等特点,在环境、临床、食品、药物检测等领域得到了广泛应用。在实际应用中,常常需要对大量复杂样品或低丰度分析物样品进行测定。而经典商品化的免疫分析方法所需时间长、样品消耗多、成本高,不能满足上述要求。因而建立快速、高通量、高灵敏和低成本的免疫检测方法已成为化学发光免疫分析方法的研究热点和发展趋势。本文简要总结近5年来化学发光免疫分析方法的研究进展及其应用,并展望其发展方向。

免疫微阵列同步多元分析系统的建立及其初步应用

本研究建立了一种基于抗体微阵列芯片的用于多个生物样品同步检测的分析方法。通过制备条件实验,确定了抗体在微阵列芯片基片表面的多项固化条件;通过鉴定及应用条件实验,确定了抗体微阵列分析系统用于三种肿瘤蛋白质检测的方法学技术指标。本分析系统体现了微阵列检测的高通量性、夹心免疫分析的高特异性和亲合素-生物素系统的高灵敏度。实验及结果分析表明,该系统具有较强的应用价值及开发潜力。

3种不同方法全自动自身抗体分析仪检测抗核抗体谱结果分析

目的探究临床常用的3种不同方法全自动自身抗体分析仪检测抗核抗体谱结果的差异。方法选取2021年4月至5月在蚌埠医学院第一附属医院确诊为系统性自身免疫病(autoimmune disease,AID)的患者72例作为AID组,留取患者相关项目检测剩余的血清样本72份,同时按照性别相同、年龄相近的原则,选取同期体检无AID的31例健康者作为对照组,留取血清样本31份。采用目前临床常用的进口全自动流式点阵发光免疫分析仪(方法A)、国产全自动流式荧光发光分析仪(方法B)和多重液相芯片免疫分析仪(方法C)分别检测两组患者血清样本中的10种抗核抗体,比较3种方法的检测阳性率,分析其阳性符合率、阴性符合率、敏感度、特异性等。为了帮助选择出对临床检验更可靠的方法学及检验设备,对3种方法进行两两比较,进行一致性评价。结果方法A、B和C与间接免疫荧光法检测的AID组阳性检出率均显著高于对照组(P<0.05)。采用方法A、B和C检测对照组10种抗体的结果均为阴性,AID组10种抗体检测结果中,方法B、C检测抗Sm抗体和抗SSA60抗体的阳性率均显著低于方法A(P<0.05),3种方法检测的其他抗体阳性率比较,差异均无统计学意义(P>0.05)。在方法A与方法B的一致性分析中,二者的平均符合率为92.22%,有1项一致性差,3项一致性一般,6项一致性好;方法A与方法C的一致性分析中,二者的平均符合率为93.89%,有5项一致性一般,5项一致性好;方法B与方法的一致性分析中,二者的平均符合率为92.65%。有1项一致性差,4项一致性一般,5项一致性好。结论目前两种国产品牌的全自动流式荧光发光分析仪和多重液相芯片免疫分析仪与进口全自动流式点阵发光免疫分析仪检测自身抗体总符合率较高,但是检测血清抗Sm抗体的一致性较差,应密切结合临床与其他实验室指标综合判断。

流式点阵免疫发光法检测抗染色质抗体在结缔组织病诊断中的价值

目的探讨流式点阵免疫发光法检测抗染色质抗体在结缔组织病诊断中的价值。方法采用流式点阵免疫荧光法检测384例结缔组织病患者[系统性红斑狼疮(SLE)243例、多肌炎(PM)39例、混合性结缔组织病(MCTD)15例、干燥综合征(SS)51例、系统性硬化症(SSc)7例、类风湿关节炎(RA)29例]、200例非风湿病患者(疾病对照组)、200名体检正常者(正常对照组)血清抗染色质抗体的表达,同时采用酶联免疫吸附试验(ELISA)检测抗双链DNA(dsDNA)抗体、抗组蛋白抗体及抗核小体抗体,采用间接免疫荧光法检测抗核抗体(ANA)。采用Spearman相关分析评估系统性红斑狼疮疾病活动度指数(SLEDIA)评分与抗染色质抗体的相关性。采用受试者工作特征(ROC)曲线分析各项指标诊断SLE的价值。结果正常对照组、疾病对照组、SLE组、MCTD组、PM组、SS组、SSc组及RA组抗染色质抗体阳性率分别为0.00%、1.00%、70.78%、73.33%、23.08%、17.65%、28.57%及10.34%。抗染色质抗体阳性率随ANA滴度的增高而增高。将MCTD组、PM组、SS组、SSc组及RA组归为非SLE组,SLE组血清抗染色质抗体水平显著高于非SLE组(P<0.0001)。SLE组中抗染色质抗体、抗dsDNA抗体、抗核小体抗体和抗组蛋白抗体中仅单个抗体阳性而其他3种抗体均阴性的比例分别为45.39%、32.34%、13.42%和7.37%,4种抗体之间均有一定比例的交叉阳性。抗核小体抗体、抗染色质抗体、抗dsDNA抗体及抗组蛋白抗体诊断SLE ROC曲线的曲线下面积(AUC)分别为0.813、0.779、0.777及0.613。抗染色质抗体与SLEDAI评分无相关性(r=0.1246,P=0.1770)。SLE患者未合并肾炎者与合并肾炎者之间血清抗染色质抗体水平差异无统计意义(P>0.05)。结论流式点阵免疫荧光法检测抗染色质抗体在结缔组织病诊断中有一定的临床价值。

液流芯片法在诊断抗中性粒细胞胞浆抗体相关性血管炎中的价值

目的探讨液流芯片法在诊断抗中性粒细胞胞浆抗体相关性血管炎(AAV)中的价值。方法选取AAV患者300例,非AAV自身免疫性疾病患者275例及健康对照者277名,分别采用线性免疫印迹法(LIA)和液相芯片法检测抗髓过氧化物酶(MPO)抗体和抗蛋白酶3(PR3)抗体,同时采用间接免疫荧光法(IFA)检测抗中性粒细胞胞浆抗体(ANCA),比较不同检测方法的性能,并筛选诊断AAV的最佳检测方案。结果LIA与液流芯片法检测抗MPO抗体、抗PR3抗体总阳性率较为一致,分别为83.1%、78.2%,kappa值均>0.75。液流芯片法的敏感性和特异性分别为85.7%、82.6%,均高于LIA的59.7%、56.6%(P<0.05)。胞浆型抗中性粒细胞胞浆抗体(cANCA)+抗PR3抗体、核周型抗中性粒细胞胞浆抗体(pANCA)+抗MPO抗体联合检测模式在AAV组中的阳性率最高,分别为42.4%、45.0%。结论液流芯片法对ANCA的检测有较高的敏感性和特异性,与IFA联合检测是诊断AAV的最佳方案。

电感耦合等离子体质谱用于多组分免疫分析研究进展

免疫分析在临床医学检测领域具有重要的地位。本课题组提出了基于电感耦合等离子体质谱(ICPMS)的免疫分析方法,利用元素标记技术结合ICP-MS检测实现多组分免疫分析。随后,研究人员在该领域做了大量研究工作,证明该方法可用于从小分子、蛋白质、核酸到细胞等一系列生物样品的检测。本文综述了基于ICP-MS免疫分析方法的特点,对其发展方向进行了展望,希望为该领域的研究工作提供参考。

多重微球流式荧光免疫技术检测抗核抗体谱的评价与临床应用

目的:分析和评估多重微球流式荧光免疫技术(MBFFI)检测抗核抗体谱的结果和临床应用价值。方法:同时用MBFFI和免疫印迹技术检测333例自身免疫病患者、44例其他疾病患者和59例健康者血清的抗核抗体谱,进行结果的一致性检验,并用间接免疫荧光技术检测抗核抗体,比较3种方法的灵敏度、特异度。结果:MBFFI与免疫印迹技术检测所有标本抗核抗体谱一致率为83. 26%,Kappa值=0. 655;病例组各项目一致率在79. 88%至99. 40%之间,Kappa值在0. 106至0. 958之间。对照组各项目一致率在96. 61%至100. 00%之间,其他疾病组各项目一致率在95. 45%至100. 00%之间。间接免疫荧光技术诊断自身免疫病的灵敏度最高(P<0. 01),MBFFI诊断自身免疫病的特异度最高(P<0. 05),MBFFI与间接免疫荧光技术联合检测可提高灵敏度(P<0. 01)。结论:多重微球流式荧光免疫技术检测抗核抗体谱与免疫印迹技术相比,一致性尚可,特异度更高,可以替代免疫印迹技术成为新一代的抗核抗体确认试验检测方法。

多指标免疫分析的研究进展

免疫分析是临床上检测生理相关蛋白质指标的主要方法.与单一指标的免疫分析相比,临床体外诊断对多指标的生理相关蛋白质免疫分析有着更为广泛的需求.通过在固相载体上完成免疫反应的非均相免疫分析具有灵敏度高的优点,是当前多指标免疫分析的主流方法.本文按照固相载体的不同,对近年来的多指标免疫分析系统分别从技术原理、实现方法、各自特点等方面的研究进展进行综合阐述.最后对比总结了不同系统的优势与不足,并展望了微流控多指标免疫分析在即时检测领域的发展前景.

Cytokines are early diagnostic biomarkers of graft-versus-host disease in liver recipients

BACKGROUND： Graft-versus-host disease （GVHD） is associated with high mortality. Early diagnosis is essential to start treatment and to improve outcomes. Because of the inflammatory nature, we hypothesis that cytokine profile of patients with GVHD may serve as diagnostic markers. The present study was to evaluate the role of cytokine profile in the diagnosis of GVHD. METHODS： An immunoassay was used to detect 29 cytokines simultaneously in the serum; the measuring sensitivity of all cytokines was pg/mL. Healthy subjects undergoing annual routine physical examinations served as negative controls; 23 patients with hepatocellular carcinoma （HCC） who had undergone liver transplantation （the LT group） comprised the test subjects. A total of 22 kidney recipients with biopsyconfirmed GVHD （the RT group） were included for comparison. HCC patients with radical surgery （the HCC group, n=22） served as positive control. The liver contents of the three cytokines, IL-2, IL-18, and IFN-γ, were detected with immunohistochemistry. Serum granzyme B and perforin were measured by flow cytometry.RESULTS： Of the 29 cytokines, the levels of IL-2 and IL-18 were increased significantly in liver recipients with GVHD compared with healthy controls （P〈0.05）. The serum levels of these three cytokines in the healthy, HCC, LT, and RT groups were IL-2： 0.90±0.02, 4.14±0.61, 5.10±0.89, and 1.48±0.09 pg/mL; IL-18： 80.61±9.35, 109.51±10.93, 230.11±12.92, and 61.98±7.88 pg/mL; IFN-γ： 24.06±3.88, 24.84±3.21, 40.37±5.88, and 15.33±4.72 pg/mL, respectively. Immunohistochemistry showed that these 3 cytokines expressions in the liver were parallel to the serum cytokine. After standard anti-GVHD treatment, the expressions of IL-2, IL-18, and IFN-y were de- creased in the liver （P〈0.05）. Serum granzyme B and perforin were significantly increased in GVHD patients （P〈0.05）. CONCLUSIONS： IL-2, IL-18 and IFN-γ were from liver and might serve as biomarkers for monitoring GVHD develop- ment and the effects of anti-GVHD treatment. Granzyme B and perforin may play a role in increasing IL-2, IL-18, and IFN-y levels in GVHD patients.

息肉状脉络膜血管病变患者血清中常见趋化因子的变化

目的检测息肉状脉络膜血管病变(PCV)患者血清中嗜酸性粒细胞活化趋化因子(Eotaxin)、生长相关蛋白(GRO-α)、白介素-8(IL-8)、干扰素诱导蛋白-10(IP-10)、单核细胞趋化蛋白-1(MCP-1)及调剂活化正常T细胞表达与分泌的趋化因子(RANTES)的表达,为进一步探讨PCV的发病机制提供依据。方法采集30例PCV患者血清,并以35例无眼底疾病的白内障患者血清作为正常对照组,应用多因子检测试剂盒对两组血清中的Eotaxin、GRO-α、IL-8、IP-10、MCP-1及RANTES的表达进行检测,分析各因子在两组血清中的表达差异。结果在PCV患者血清中Eotaxin平均浓度为(48.90±24.76)pg/m L,高于对照组(37.49±14.39)pg/m L,差异具有统计学意义(P<0.05)。其他几种趋化因子的表达无明显差异(P>0.05)。结论 Eotaxin在PCV血清中表达明显增高,提示Eotaxin可能在PCV的发生和发展中起重要作用。

Changes in Biomarkers after Initial Periodontal Treatment in Gingival Crevicular Fluid from Patients with Chronic Periodontitis Presenting with Drug-Induced Gingival Overgrowth

Calcium channel blocker-induced gingival overgrowth (CCB-GO) is increasing in elderly patients who have been prescribed medication for hypertension for years. The purpose of the present study was to analyze the comprehensive protein expression levels of candidate biomarkers in gingival crevicular fluid (GCF) from CCB-GO patients. Eleven GO patients (10 males and one female, mean ± SD: age: 64.4 ± 14.0 years) who had been systemically prescribed CCBs, either amlodipine or nifedipine, for hypertension for at least 12 months were recruited. Before (baseline) and 4 weeks after initial periodontal treatments, subgingival plaque and GCF samples were taken from two sites per patient: sites affected by CCB-GO and chronic periodontitis. Measurement of clinical parameters and quantitative analysis of periodontopathic bacteria using real-time PCR were performed. Biomarkers/cytokines in GCF were examined using multiplex bead immunoassays. The Mann-Whitney U test was used to compare the collected data between groups. The correlations between pairs of biomarkers were assessed using the Spearman correlation relationship. Levels of two of the 14 biomarkers, interleukin (IL)-1β and transforming growth factor (TGF)- β, were significantly decreased in CCB-GO sites after initial periodontal therapy. The intragroup comparison at baseline showed that counts of Treponema denticola in the GO group were significantly higher than those in the chronic periodontitis group (P β and TGF-β in CCB-GO patients. These factors are involved in initiation and progression of GO as well as periodontitis.

全自动流式点阵免疫发光法测定抗MPO和抗PR3抗体的性能评估

目的评估全自动流式点阵免疫发光法(MBIA法)测定抗髓过氧化物酶(MPO)和抗蛋白酶3(PR3)抗体的检测性能。方法按照美国临床实验室标准化委员会(CLIS)EP15-A3文件进行精密度验证,按照EP28-A3文件验证参考区间。比较MBIA法和ELISA法检测抗PR3和抗MPO结果,结合间接免疫荧光法(IFT)评价不同方法对抗中性粒细胞胞浆抗体(ANCA)相关性血管炎(AAV)的诊断效能。结果 MBIA法检测抗PR3和MPO抗体批内、批间不精密度分别为3.65%、4.10%和2.28%、3.32%,小于厂家声明允许不精密度批内4.70%、批间8.60%,符合要求。MBIA和ELISA两种方法检测抗PR3和抗MPO的阳性、阴性、总符合率分别是40.00%、90.89%、90.72%和80.49%、97.65%、96.10%。MBIA法和ELISA法的敏感性、特异性以及阳性预测值分别是53.85%、91.69%、67.31%和55.38%、79.71%、46.45%。与IFT结果结合后,阳性预测值可分别提高至83.56%、73.48%。结论 MBIA法检测性能符合要求,其诊断AAV的效能要优于ELISA法,与IFT法结果结合可提高对AAV诊断的阳性预测值。

Point-of-Care Immunoassays with Tunable Detection Range for Detecting Infection in Intensive Care Unit

One of the difficulties of a multiplexed analytical assay is matching the concentration range of different markers.Here,the authors develop an automatic,multiplexed point-of-care(POC)immunoassay that allows detecting C-reactive protein(CRP),procalcitonin(PCT),and interleukin 6(IL-6)in 50μL serum samples with limits of detection 1.87μg/mL,0.17 ng/mL,and 49.75 pg/mL,respectively.The authors use electrospun fibers and surface chemistry of gold nanoparticles(AuNPs)to adjust the detection range of different biomarkers.The authors have proposed some new diagnostic indicators(mScore and mScorePlus)that combine the results of CRP,PCT,IL-6,and complete blood counts(CBCs).The authors also tracked changes in CRP,PCT,and IL-6 of patients in an intensive care unit(ICU).The authors find that mScore and mScorePlus have advantages in improving the diagnostic accuracy and providing more analytical information.mScore and mScorePlus are effective tools to detect infections,differentiate bacterial and viral infections,and monitor disease.Integrating multiple markers into one straightforward parameter is an effective method for analytical applications.The authors believe that this method provides a general way for chemists to develop increasingly accurate detection methods and indicators.

多重生物检测技术研究进展

多重生物检测因其快速、耗费少、所需样品量少等优点,被广泛用于细菌、病毒、真菌、寄生虫及自身免疫相关抗体等抗原或抗体的快速筛查诊断。多重核酸检测的新技术包括巢式PCR(nPCR)、多重串联PCR、基于毛细管电泳的GeXP平台和luminex xMAP多重分析技术平台,多重免疫检测的新技术包括基于上转换发光、多线点、表面等离子共振技术的多重生物检测及基于蛋白芯片的阵列array-ELISA检测。本文综述了近年来多重生物检测新技术在呼吸道感染、腹泻、自身免疫性缺陷和肿瘤等疾病中的应用进展。