基于双分子信标对猪圆环病毒Ⅱ型的简单快速检测

根据猪圆环病毒(PCV2)基因组为单链DNA的特点,设计了两条可与PCV2基因组序列特异性杂交的分子信标,建立了基于双分子信标法检测PCV2的方法。实验结果表明,双分子信标法比单分子信标法的灵敏度更高。在10 mmol/L MgCl2、20 mmol/L Tris-HCl(pH=8.0)的杂交缓冲溶液,40℃孵育30 min的优化检测体系下,此方法可实现对检测样本在2~200 nmol/L线性范围内的检测,检出限可达1 nmol/L。将双分子信标检测体系用于18例可疑猪瘟样品病毒检测,其中8例呈PCV2阳性,检测结果与PCR法一致,证明此双分子信标法可用于实际猪感染PCV2的诊断。     

一种基于分子信标荧光探针快速检测烟草花叶病毒的新方法

研究了一种利用新型荧光探针——分子信标进行植物病毒检测的方法，可以不要求对病毒RNA进行严格的分离和纯化，就能快捷地得到检测结果。此方法被用于烟草花叶病毒(tobacco mosaic virus，TMV)基因组RNA的检测，为植物病毒分子生物学的研究提供了新的手段。     

基于磁致伸缩材料的猪瘟病毒无线免疫传感器

通过在磁致伸缩材料表面形成金-硫键自组装膜固载猪瘟多抗,构建了一种新型、灵敏、便携、低成本的猪瘟病毒无线免疫传感器。该传感器基于抗原抗体特异性结合使猪瘟病毒附着在磁致伸缩材料表面从而导致磁弹片共振频率降低的原理,通过测定共振频率偏移量实现对猪瘟病毒的无线检测。采用扫描电子显微镜(SEM)对抗体修饰及猪瘟病毒检测过程进行了表征,利用该传感器对不同浓度的猪瘟病毒进行检测,线性范围为0.5~2.5μg/m L,响应灵敏度为65 Hz/(μg·m L~(-1)),检出限为0.5μg/m L。     

新冠病毒临床检测中的生化技术和原理*

新型冠状病毒肺炎爆发以来,如何准确快速地识别感染者是疫情防控的关键。本文系统地介绍已用于新冠病毒临床检测的各类技术,对涉及的生化原理进行深入浅出的阐述,旨在让广大群众更好地了解新冠病毒检测的方法和原理,积极配合医务工作者的工作,增强战胜新冠疫情的信心。     

生物传感器在新冠病毒检测中的应用

自新型冠状病毒肺炎(COVID-19)爆发以来,准确高效、快速便捷的新型冠状病毒(SARS-CoV-2)筛查越来越在疫情防控中体现出其重要性.传统的检测方法无法满足短时间内SARS-CoV-2大规模感染时的检测需求.生物传感技术具有灵敏度高、选择性好、低成本、易于微型化等优点,其检测时间短的优势还可用于开发即时检测设备,是临床诊断中实时检测SARS-CoV-2的潜在替代方案.本综述简要阐述了光学生物传感器、电化学生物传感器、可穿戴式生物传感器、磁性生物传感器、金纳米颗粒生物传感器以及适配体生物传感器的构建方法、工作原理,并总结了其在新冠病毒检测领域的最新应用.最后本综述对生物传感器在新冠病毒检测领域的技术瓶颈与未来的发展趋势进行了总结与讨论.     

食物过敏原免疫学检测技术研究进展

免疫学检测方法因特异性强、灵敏度高和操作简便的优点而备受青睐.该文介绍了部分国家和地区食物过敏原标签标注情况,食物过敏原免疫学检测技术,包括酶联免疫吸附法(ELISA)、免疫层析技术、生物传感器技术的研究进展和应用.通过与质谱、链式聚合酶反应技术(PCR)对比,探讨了食物过敏原免疫学检测技术的发展方向.     

河豚毒素的样品前处理与快速检测技术研究进展

河豚毒素是一种毒性极强的小分子、非蛋白神经毒剂,其性质相对稳定,缺乏特效解毒剂。其检测技术主要有生物检测法、理化检测法和免疫学检测法,其中免疫检测因快速、简便、灵敏而得到较为广泛的应用。该文综述了河豚毒素的样品前处理技术、免疫学快速检测技术及相关专利的研究进展情况,并对河豚毒素检测产品的开发进行了展望。     

基于免疫磁球的埃博拉病毒糖蛋白高灵敏比色检测

用免疫磁球捕获埃博拉病毒糖蛋白,与生物素化抗体形成免疫夹心复合物,然后链霉亲和素标记辣根过氧化物酶(SA-HRP)催化3,3',5,5'-四甲基联苯胺(TMB)进行显色,通过370 nm处吸光度对病毒糖蛋白进行定量。对免疫磁球进行了免疫荧光表征,通过对照实验验证方法的可靠性,并对检测条件进行了优化。结果表明,吸光度与病毒糖蛋白浓度在1.0~25.0 ng/m L内呈线性关系,检出限达到0.18 ng/m L。该方法重现性较好,特异性好,抗干扰能力强,可实现复杂样品中埃博拉病毒的检测。     

高灵敏度分析检测方法在病毒研究中的应用

病毒与人类息息相关,病毒的分析与检测一直受到人们的关注。本文主要评述了化学发光法、荧光光谱法和原子力显微镜成像法的工作原理以及它们在病毒研究中的具体应用,为病毒的成像、检测以及组分分析提供了可行的技术手段,并对其的发展作了展望。     

G-四链体探针在病毒基因检测和成像中的应用进展

病毒依赖宿主细胞完成生命周期,影响着地球上所有生物,是人类疾病的主要原因之一,对人类健康以及社会经济造成严重影响,因此开发检测病毒的工具对防治病毒感染和预防病毒导致的疾病具有重要意义.病毒基因组在病毒的生命周期中发挥着重要作用,为病毒检测提供了有效靶点.G-四链体(G4)是由富含鸟嘌呤的核酸折叠形成的一种稳定的核酸二级...     

Prediction of African Swine Fever Virus Inhibitors by Molecular Docking-Driven Machine Learning Models

African swine fever virus (ASFV) causes a highly contagious and severe hemorrhagic viral disease with high mortality in domestic pigs of all ages. Although the virus is harmless to humans, the ongoing ASFV epidemic could have severe economic consequences for global food security. Recent studies have found a few antiviral agents that can inhibit ASFV infections. However, currently, there are no vaccines or antiviral drugs. Hence, there is an urgent need to identify new drugs to treat ASFV. Based on the structural information data on the targets of ASFV, we used molecular docking and machine learning models to identify novel antiviral agents. We confirmed that compounds with high affinity present in the region of interest belonged to subsets in the chemical space using principal component analysis and k-means clustering in molecular docking studies of FDA-approved drugs. These methods predicted pentagastrin as a potential antiviral drug against ASFVs. Finally, it was also observed that the compound had an inhibitory effect on AsfvPolX activity. Results from the present study suggest that molecular docking and machine learning models can play an important role in identifying potential antiviral drugs against ASFVs.     

Recombination and expression of classical swine fever virus (CSFV) structural protein E2 gene in Chlamydomonas reinhardtii chroloplasts

The expression of classical swine fever virus (CSFV) structural protein E2 in different vectors, which has been shown to carry critical epitopes, has been established. Here, we reported a Chlamydomonas reinhardtii chloroplast expression vector, P64E2, containing classical swine fever virus structural protein E2 gene, which was constructed and transferred to C. reinhardtii by biolistic bombardment method. The transformants were identified by PCR, Southern blotting, Western blotting after selecting on resistant media. ELISA quantification assay showed that the expressed E2 protein accumulated up to 1.5-2% of the total soluble protein. The results of the study on the immunological activity indicated that the protein E2 expressed in C. reinhardtii chloroplasts could elicit animal bodies to produce antibodies against protein E2.     

Rapid separation and identification of the subtypes of swine and equine influenza A viruses by electromigration techniques with UV and fluorometric detection

Influenza A is viral disease, which is a cause of yearly epidemics and, potentially, pandemics. The conventional techniques used today are equipment-demanding, time-consuming and laborious. Recently, we have confirmed that the capillary isoelectric focusing is a suitable fast alternative for the verifying of virus purity. In the wide pH gradient of pH range 2.0-7.5 the isoelectric points for subtypes of equine (H3N8) and swine (H1N2) influenza A viruses were determined approximately as 6.6 and 6.5, respectively. In this contribution we have verified these findings using different isolates of different viral subtypes of swine influenza, H1N1, H1N2, and of equine influenza, H3N8, H7N7, which were separated by capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF) in the narrow pH gradient pH range from 6.0 to 7.0. It was found that the isoelectric points of different isolates and subtypes of equine and swine influenza are almost independent of their origin. The electromigration velocities of subtypes of equine or swine influenza viruses were dependent on the antigenic subtypes of their surface glycoproteins. The detection sensitivity of the influenza viruses labeled by the fluorescent non-ionogenic tenside based on poly(ethylene glycol)pyrenebutanoate for fluorometric detection was increased and down to ten labeled viruses were detected. The isoelectric points of the native and labeled equine and swine influenza A viruses and their subtypes do not differ. According to our experiments these methods appear to be useful for the fast preliminary differentiation of influenza viruses in future.     

Laboratory tests for human immunodeficiency viruses.

Two types of human immunodeficiency viruses are known. Both type 1 (HIV-1) and type 2 (HIV-2) can lead to AIDS. This article describes laboratory tests that are used for diagnosis of HIV infection and for monitoring disease progression or the effects of therapy. These tests are based on detection of host antibodies, viral antigens, viral nucleotide sequences, or cultivation of the virus in vitro. The principles underlying test methods for differentiation between HIV-1 and HIV-2 are described.     

Extracellular vesicles based electrochemical biosensors for detection of cancer cells: A review

Extracellular vesicles (EVs) derived from cancer cells are considered as ideal biomarker for liquid biopsy in cancer diagnosis, and are stable and abundant. Electrochemical methods for the detection of EVs are preferred over conventional methods such as Western blotting and enzyme-linked immunosorbent assay for their high sensitivity and real-time detection. This article summaries studies proposing the electrochemical methods utilizing immunological and molecular methodologies for detecting EVs derived biomacromolecules such as miRNAs and transmembrane protein for cancer diagnosis. Moreover, the electrochemical detection methods are compared and future prospects for the development of electrochemical methods for EVs detection are concluded.     

Electrochemical sensing of trypanosome- and flavivirus-related neglected tropical diseases

Virus infections and parasitic diseases that are often found in tropical countries represent a large burden on societies in those regions but are unfortunately rarely studied by electrochemistry. These neglected tropical diseases (NTDs) are a concerning issue, especially in developing nations because a challenged healthcare system often negatively affects the economy. Besides the exploration of new treatment strategies, the development of early diagnostic methods is crucial to attenuate negative impacts related to NTDs. This review highlights the most recent developments of electrochemical biosensing efforts for five NTDs, Chagas disease, leishmaniasis, sleeping sickness, dengue fever and Zika virus infection, which affect a number of Latin American, African, and Asian countries. Special focus is placed on the design and fabrication of genosensors and immunosensors, designed as innovative and promising diagnostic tools with the potential of being integrated into portable systems to advance the development of point-of-care devices.     

Magnetic beads-based electrochemical immunosensor for detection of pseudorabies virus antibody in swine serum

A novel magnetic electrochemical immunosensor has been developed for the detection of pseudorabies virus antibody in swine serum. The magnetic glass carbon electrode was fabricated to manipulate magnetic beads for the direct sensing applications. Magnetic beads were employed as the platforms for the immobilization and immunoreaction process, and gold nanoparticles were chosen as electroactive labels for the electrochemical detection. The parameters concerning the assay strategy were carefully investigated. Under the optimal conditions, the linear response range of pseudorabies virus antibody dilution ratio (standard positive serum) was 1:250 to 1:1000 with a detection limit of 1:1000. Finally, this developed immunoassay method was successfully applied in the detection of pseudorabies virus antibody in swine serum, and had a good diagnostic accordance in comparison with ELISA.     

Rapid screening swine foot-and-mouth disease virus using micro-ELISA system

In order to tackle both regional and global foot-and-mouth disease virus (FMDV) epdimics, we hereby develop a rapid microfluidic thermal lens microscopic method to screen swine type O FMDV with good efficiency. The scheme has great merits in terms of field portability, sample volume, assay time, analytical sensitivity, and test reproducibility.     

Evaluation of the antiviral and antimicrobial activities of triterpenes isolated from Euphorbia segetalis

A phytochemical reinvestigation of the whole plant of Euphorbia segetalis yielded five tetracyclic triterpenes: 3beta-hydroxy-cycloart-25-en-24-one (1), cycloart-25-ene-3beta,24-diol (2), cycloart-23-ene-3beta,25-diol (3), lanosta-7,9(11),24-trien-3beta-ol (4) and lanosta-7,9(11),24(31)-trien-3beta-ol (5). beta-acetoxy-cycloart-25-en-24-one (1a) and glutinol (6), lupenone (7), dammaranodienol (9), cycloartenol acetate (10), 24-methylenecycloartanol acetate (11) and beta-sitosterol (12), isolated previously, were evaluated for their antiviral activities against Herpes simplex virus (HSV) and African swine fever virus (ASFV). Lupenone exhibited strong viral plaque inhibitory effect against HSV-1 and HSV-2. The in vitro antifungal and antibacterial activities of la, cycloart-23-ene-3beta,25-diol, 3-acetate (3a) and 6-12 were also investigated.