ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Ruijin Zeng,Minghao Qiu,Qing Wan,Zhisheng Huang,Xiaolong Liu,Dianping Tang,Dietmar Knopp
Large-scale, rapid, and inexpensive serological diagnoses of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) are of great interest in reducing virus transmission at the population level; however, their development is greatly plagued by the lack of available point-of-care methods, leading to low detection efficiency. Herein, an ultrasensitive smartphone-based electrochemical immunoassay is reported for rapid (less than 5 min), low-cost, easy-to-implement detection of the SARS-CoV-2 nucleocapsid protein (SARS-CoV-2 N protein). Specifically, the electrochemical immunoassay was fabricated on a screen-printed carbon electrode coated with electrodeposited gold nanoparticles, followed by incubation of anti-N antibody (Ab) and bovine serum albumin as the working electrode. Accompanied by the antigen–antibody reaction between the SARS-CoV-2 N protein and the Ab, the electron transfer between the electroactive species [Fe(CN)6]3–/4– and the electrode surface is disturbed, resulting in reduced square-wave voltammetry currents at 0.075 V versus the Ag/AgCl reference electrode. The proposed immunoassay provided a good linear range with SARS-CoV-2 N protein concentrations within the scope of 0.01–1000 ng/mL (R2 = 0.9992) and the limit of detection down to 2.6 pg/mL. Moreover, the detection data are wirelessly transmitted to the interface of the smartphone, and the corresponding SARS-CoV-2 N protein concentration value is calculated and displayed. Therefore, the proposed portable detection mode offers great potential for self-differential diagnosis of residents, which will greatly facilitate the effective control and large-scale screening of virus transmission in resource-limited areas.
研究领域:
医学诊断
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生物传感器
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病毒学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Ruimin Wang,Shuai Wang,Wanze Guo,Tiantian Zhang,Qing Kang,Pengcheng Wang,Feimeng Zhou,Lixia Yang
A thin-layer flow cell of low internal volume (12 μL) is incorporated in a flow injection analysis (FIA) system for simultaneous and real-time photoelectrochemical (PEC) immunoassay of anti-SARS-CoV-2 spike 1 (S1) and anti-SARS-CoV-2 nucleocapsid (N) antibodies. Covalent linkage of S1 and N proteins to two separate polyethylene glycol (PEG)-covered gold nanoparticles (AuNPs)/TiO 2 nanotube array (NTA) electrodes affords 10 consecutive analyses with surface regenerations in between. An indium tin oxide (ITO) allows visible light to impinge onto the two electrodes. The detection limits for anti-S1 and anti-N antibodies were estimated to be 177 and 97 ng mL −1 , respectively. Such values compare well with those achieved with other reported methods and satisfy the requirement for screening convalescent patients with low antibody levels. Additionally, our method exhibits excellent intra-batch (RSD = 1.3%), inter-batch (RSD = 3.4%), intra-day (RSD = 1.0%), and inter-day (RSD = 1.6%) reproducibility. The obviation of an enzyme label and continuous analysis markedly decreased the assay cost and duration, rendering this method cost-effective. The excellent anti-fouling property of PEG enables accuracy validation by comparing our PEC immunoassays of patient sera to those of ELISA . In addition, the simultaneous detection of two antibodies holds great potential in disease diagnosis and immunity studies.
研究领域:
医学诊断
·
生物传感器
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免疫学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 4
·1-13
·
IF 5.3
Li Yang,Long Feng,Han Shitong,Wang Yi,Zhu Anna
Timely and accurate detection of the virus is of great significance to prevent the virus’s harm and control the epidemic. Here, an aptasensor based on the principle of promoting hybridization through aptamer conformational change was designed to quantitatively detect the spike (S) protein of SARS-CoV-2. When the S protein binds to the 3' end of the aptamer, the 5' end of the aptamer tansforms into a straight hybridization region, which will greatly facilitate the hybridization with complementary DNA (cDNA). In the absence of S protein, hybridization is less likely to occur due to the complex G-quadruplex structure of aptamer. According to this principle, cDNA is modified onto magnetic beads (MBs) or onto the optical fiber probe of an evanescent wave fluorescence aptasensor (EWFA) detection platform to capture the fluorescently labeled aptamer-S protein conjugate, two kinds of quantitative detection methods for SARS-CoV-2 S protein were established. In particular, simple, rapid and sensitive detection could be obtained based on the EWFA detection platform, in which the whole detection procedure including the measurement and regeneration takes only 14 min, the LOD is 5.34 ng/mL, the linear response range is 141.49 to 9507.36 ng/mL, and the optical fiber probe could be reused for 19 times. The EWFA detection platform is also potentially applicable to detect other protein biomarkers only by replacing the specifically modified optical fiber probes. Graphical
研究领域:
生物化学
·
分子诊断
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病毒学
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产品关联
DOI
PubMed
高影响因子
ACS Applied Bio Materials
·
2024
·
IF 4.6
Hao Chen,Xilin Xiong,Yuan Huang,Bo Huang,Xinxin Luo,Qi Ke,Pengyu Wu,Suxiao Wang
The global outbreak of the COVID-19 pandemic has indisputably wreaked havoc on societies worldwide, compelling the scientific community to seek urgently needed therapeutic agents with low-cost and low-side effect profiles. Numerous approaches have been investigated in the quest to prevent or treat COVID-19, but many of them exhibit unwelcome side effects, such as dysfunctional viral immune responses and inflammation. Herein, we present the preparation of solid natural human pulmonary alveolar epithelial cell (ATII) membrane-coated PLGA NPs (PLGA NPs@ATII-M), which demonstrate remarkable affinity and competitiveness to neutralize the SARS-CoV-2 S1 protein-coated NPs (SCMMA NPs–S1), which are employed as a surrogate for coronavirus particles. In addition, we first considered the antifouling properties of these types of NPs, and we found that this membrane-coated NP formulation boasts excellent antifouling capabilities, which serve to protect their neutralization properties out of shielding by protein coronas in blood circulation. Moreover, this formulation is easily prepared and stored with a low-cost profile and exhibits good specificity, high targeting efficiency, and potentially side effect avoiding, thus making it a highly promising candidate for COVID-19 treatment.
研究领域:
药学
·
纳米技术
·
病毒学
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
MOLECULAR PHARMACEUTICS
·
2024
·
IF 4.5
Ningze Zheng,Shurui Liu,Jianheng Chen,Yue Xu,Wenyin Cao,Jinyi Lin,Guang Lu,Guigen Zhang
The development of biomolecule delivery systems is essential for the treatment of various diseases such as cancer, immunological diseases, and metabolic disorders. For the first time, we found that SARS-CoV-2-encoded nonstructural protein 2 (NSP2) can be secreted from the cells, where it is synthesized. Brefeldin A and H89, inhibitors of ER/Golgi secretion pathways, did not inhibit NSP2 secretion. NSP2 is likely secreted via an unconventional secretory pathway. Moreover, both secreted and purified NSP2 proteins were able to traverse the plasma membrane barrier and enter both immortalized human umbilical vein endothelial cells and tumor cell lines. After entry, the NSP2 protein was localized in only the cytoplasm. Cytochalasin D, a potent inhibitor of actin polymerization, inhibited the entry of NSP2. NSP2 can carry other molecules into cells. Burkholderia lethal factor 1, a monomeric toxin from the intracellular pathogen Burkholderia pseudomallei, has demonstrated antitumor activity by targeting host eukaryotic initiation translation factor 4A. An NSP2-BLF1 fusion protein was translocated across the cellular membranes of Huh7 cells and mediated cell killing. By using different approaches, including protein purification, chemical inhibition, and cell imaging, we confirm that NSP2 is able to deliver heterologous proteins into cells. NSP2 can act as a potential delivery vehicle for proteins.
研究领域:
分子生物学
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细胞生物学
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癌症研究
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药理学
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DOI
PubMed
高影响因子
BRIEFINGS IN BIOINFORMATICS
·
2021
·Vol. 22
·期 2
·1378-1386
·
IF 11.622
Li Shasha,Liu Wenli,Chen Yangzhen,Wang Liqin,An Wenlin,An Xiaoping,Song Lihua,Tong Yigang,Fan Huahao,Lu Chenyang
Antiviral therapies targeting the pandemic coronavirus disease 2019 (COVID-19) are urgently required. We studied an already-approved botanical drug cepharanthine (CEP) in a cell culture model of GX_P2V, a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-related virus. RNA-sequencing results showed the virus perturbed the expression of multiple genes including those associated with cellular stress responses such as endoplasmic reticulum (ER) stress and heat shock factor 1 (HSF1)-mediated heat shock response, of which heat shock response-related genes and pathways were at the core. CEP was potent to reverse most dysregulated genes and pathways in infected cells including ER stress/unfolded protein response and HSF1-mediated heat shock response. Additionally, single-cell transcriptomes also confirmed that genes of cellular stress responses and autophagy pathways were enriched in several peripheral blood mononuclear cells populations from COVID-19 patients. In summary, this study uncovered the transcriptome of a SARS-CoV-2-related coronavirus infection model and anti-viral activities of CEP, providing evidence for CEP as a promising therapeutic option for SARS-CoV-2 infection.
研究领域:
病毒学
·
分子生物学
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药理学
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转录组学
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传染病学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Haochen Qi,Zhiwen Hu,Zhongliang Yang,Jian Zhang,Jie Jayne Wu,Cheng Cheng,Chunchang Wang,Lei Zheng
The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has lasted for almost 2 years. Stemming its spread has posed severe challenges for clinical virus detection. A long turnaround time, complicated operation, and low accuracy have become bottlenecks in developing detection techniques. Adopting a direct antigen detection strategy, we developed a fast-responding and quantitative capacitive aptasensor for ultratrace nucleocapsid protein detection based on a low-cost microelectrode array (MEA) chip. Employing the solid–liquid interface capacitance with a sensitivity of picofarad level, the tiny change on the MEA surface can be definitively detected. As a result, the limit of detection reaches an ultralow level of femtogram per milliliter in different matrices. Integrated with efficient microfluidic enrichment, the response time of this sensor from the sample to the result is shortened to 15 s, completely meeting the real-time detection demand. Moreover, the wide linear range of the sensor is from 10–5 to 10–2 ng/mL, and a high selectivity of 6369:1 is achieved. After application and evaluation in different environmental and body fluid matrices, this sensor and the detection method have proved to be a label-free, real-time, easy-to-operate, and specific strategy for SARS-CoV-2 screening and diagnosis.
研究领域:
生物医学工程
·
病毒学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
Nature Biomedical Engineering
·
2025
·
IF 26.6
Li Cuifeng,Li Yuxin,Zhou Bini,Li Tong,Wei Xiaohui,Chen Kun,Chen Wenlong,Shi Ziwei,Dai Xiaobin,Zhang Jiachen,Yang Chen,Ji Zhenglin,Sun Wenbo,Gao Ji,Wu Jing,Zhao Binbin,Min Xin,Li Yujie,Lin Lin,Yang Wei,Wang Min,Liu Zezhong,Liu Yaping,Zhu Chenyou,Yang Bo,Xu Jiang-Fei,Yan Li-Tang,Shi Yi,Lu Lu,Zhang Linqi,Ding Qiang,Xue Jing,Hou Baidong,Qi Hai,Liu Jiangning,Yang Yuhe R.,Liu Dongsheng,Liu Wanli
The emergence of novel infectious disease has intensified demand for more advanced vaccine development and more potent adjuvants to enhance immunogenicity. Here we introduce a dynamic DNA supramolecular matrix assembled from five unmodified, short DNA single strands, serving as a safe, multifaceted adjuvant platform. This DNA matrix elicits a robust humoral response with minimal adverse effects, generating potent neutralizing antibodies and conferring robust protection against SARS-CoV-2 and Streptococcus pneumoniae infections. Its dynamic colloidal feature prolongs the in vivo retention of both DNA and antigen, facilitating lymphatic-targeted transportation and presentation. This process leads to a robust pro-inflammatory response in both the vaccinated site and draining lymph node, which, in turn, promotes the recruitment and activation of immune cells, leading to a rapid, effective antigen-specific antibody response. The enhanced function of DNA matrix depends on the canonical TLR9–MyD88 signalling axis in dendritic cells. In addition, only right-handed, not left-handed, chirality of the DNA strands forms d -DNA matrix and promotes immune activations. Thus, this DNA matrix functions as an all-in-one adjuvant platform, opening promising avenues for future vaccine design.
研究领域:
免疫学
·
微生物学
·
生物材料
·
疫苗学
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产品关联
DOI
PubMed
高影响因子
Gut Microbes
·
2022
·
IF 9.434
Liqiu Jia,Shufeng Weng,Jing Wu,Xiangxiang Tian,Yifan Zhang,Xuyang Wang,Jing Wang,Dongmei Yan,Wanhai Wang,Fang Fang,Zhaoqin Zhu,Chao Qiu,Wenhong Zhang,Ying Xu,Yanmin Wan
The origins of preexisting SARS-CoV-2 cross-reactive antibodies and their potential impacts on vaccine efficacy have not been fully clarified. In this study, we demonstrated that S2 was the prevailing target of the preexisting S protein cross-reactive antibodies in both healthy human and SPF mice. A dominant antibody epitope was identified on the connector domain of S2 (1147-SFKEELDKYFKNHT-1160, P144), which could be recognized by preexisting antibodies in both human and mouse. Through metagenomic sequencing and fecal bacteria transplant, we demonstrated that the generation of S2 cross-reactive antibodies was associated with commensal gut bacteria. Furthermore, six P144 reactive monoclonal antibodies were isolated from naïve SPF mice and were proven to cross-react with commensal gut bacteria collected from both human and mouse. A variety of cross-reactive microbial proteins were identified using LC-MS, of which E. coli derived HSP60 and HSP70 proteins were confirmed to be able to bind to one of the isolated monoclonal antibodies. Mice with high levels of preexisting S2 cross-reactive antibodies mounted higher S protein specific binding antibodies, especially against S2, after being immunized with a SARS-CoV-2 S DNA vaccine. Similarly, we found that levels of preexisting S2 and P144-specific antibodies correlated positively with RBD binding antibody titers after two doses of inactivated SARS-CoV-2 vaccination in human. Collectively, our study revealed an alternative origin of preexisting S2-targeted antibodies and disclosed a previously neglected aspect of the impact of gut microbiota on host anti-SARS-CoV-2 immunity.
研究领域:
免疫学
·
微生物学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Xiaohu Liu,Ruifeng Xie,Kang Li,Zhelei Zhu,Xi Huang,Qian He,Zhe Sun,Haiyang He,Yuancai Ge,Qingwen Zhang,Hu Chen,Yi Wang
We have developed a convenient surface-enhanced Raman scattering (SERS) platform based on vertical standing gold nanowires (v-AuNWs) which enabled the on-mask detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) related substances such as the Spike-1 protein and the corresponding pseudo-virus. The Spike-1 protein was clearly distinguished from BSA protein with an accuracy above 99 %, and the detection limit could be achieved down to 0.01 μg/mL. Notably, a similar accuracy was achieved for the pseudo-SARS-CoV-2 (pSARS-2) virus as compared to the pseudo-influenza H7N9 (pH7N9) virus. The sensing strategy and setups could be easily adapted to the real SARS-CoV-2 virus and other highly contagious viruses. It provided a promising way to screen the virus carriers by a fast evaluation of their wearing v-AuNWs integrated face-mask which was mandatory during the pandemic.
研究领域:
分析化学
·
纳米技术
·
病毒学
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Signal Transduction and Targeted Therapy
·
2024
·Vol. 9
·期 1
·1-13
·
IF 40.8
Yang Minghui,Li Chunhui,Ye Guoguo,Shen Chenguang,Shi Huiping,Zhong Liping,Tian Yuxin,Zhao Mengyuan,Wu Pengfei,Hussain Abid,Zhang Tian,Yang Haiyin,Yang Jun,Weng Yuhua,Liu Xinyue,Wang Zhimin,Gan Lu,Zhang Qianyu,Liu Yingxia,Yang Ge,Huang Yuanyu,Zhao Yongxiang
Emerging and recurrent infectious diseases caused by human coronaviruses (HCoVs) continue to pose a significant threat to global public health security. In light of this ongoing threat, the development of a broad-spectrum drug to combat HCoVs is an urgently priority. Herein, we report a series of anti-pan-coronavirus ssDNA aptamers screened using Systematic Evolution of Ligands by Exponential Enrichment (SELEX). These aptamers have nanomolar affinity with the nucleocapsid protein (NP) of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and also show excellent binding efficiency to the N proteins of both SARS, MERS, HCoV-OC43 and -NL63 with affinity K D values of 1.31 to 135.36 nM. Such aptamer-based therapeutics exhibited potent antiviral activity against both the authentic SARS-CoV-2 prototype strain and the Omicron variant (BA.5) with EC 50 values at 2.00 nM and 41.08 nM, respectively. The protein docking analysis also evidenced that these aptamers exhibit strong affinities for N proteins of pan-coronavirus and other HCoVs (−229E and -HKU1). In conclusion, we have identified six aptamers with a high pan-coronavirus antiviral activity, which could potentially serve as an effective strategy for preventing infections by unknown coronaviruses and addressing the ongoing global health threat.
研究领域:
病毒学
·
分子生物学
·
药物开发
·
传染病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Haodong Cui,Wenxing Song,Xiaoling Ru,Wen Fu,Ling Ji,Wenhua Zhou,Zhen Zhao,Guangbo Qu,Xue-Feng Yu,Guibin Jiang
The ongoing outbreak of the novel coronavirus disease 2019 (COVID-19) draws worldwide concerns due to its long incubation period and strong infectivity. Although RT-PCR-based methods are being widely applied for clinical diagnosis, timely and accurate diagnosis towards COVID-19 causing virus, the SARS-CoV-2, is still limited due to labor-intensive and time-consuming operations. Herein, we report a new viral RNA extraction method based on poly-(amino ester) with carboxyl group (PC)-coated magnetic nanoparticles (pcMNPs) for the sensitive detection of SARS-CoV-2. This method combines the lysis and binding steps into one step, and refines multiple washing steps into one step, giving a turnaround time of less than 9 min. Furthermore, the extracted pcMNP-RNA complexes can be directly introduced into subsequent RT-PCR reactions without elution. This simplified viral RNA method could be well adapted in fast manual and automated high-throughput nucleic acids extraction protocols suitable for different scenarios. A high sensitivity down to 100 copies/mL and a linear correlation between 100 and 10 6 copies/mL of SARS-CoV-2 pseudovirus particles are achieved in both protocols. Benefitting from the simplicity and excellent performances, this new method can dramatically improve the efficiency and reduce operational requirements for the early clinical diagnosis and large-scale SARS-CoV-2 nucleic acid screening.
研究领域:
分子生物学
·
医学诊断
·
纳米技术
·
传染病研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）