Virology Journal
·
2025
·
IF 3.8
Jinlei Guo,Yunfeng Shan,Ge Hu,Xiaojuan Zhu,Kangchen Zhao,Tao Wu,Qiao Qiao,Ying Chi,Lunbiao Cui,Yiyue Ge
Background Monkeypox virus (MPXV) infection has garnered significant global attention due to its rising incidence and substantial public health implications. A rapid, sensitive, and accurate diagnostic method is urgently required to enable early intervention and effective management of MPXV outbreaks. Results In this study, we developed a novel one-step assay that integrates loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12b in one-pot for the detection of MPXV. The entire detection process did not require opening the lid of the reaction tube and could be completed within 40 min using extracted viral nucleic acids, which is faster than real-time quantitative PCR (qPCR). And the results could be interpreted through either real-time fluorescence or naked-eye visualization. The limit of detection (LOD) of the assay was demonstrated to be 6.5 copies per reaction and no cross-reactivity with other pathogens such as HSV, EBV, CVA16, EV-A71, and MV was found. Furthermore, when evaluated using 113 clinical samples, the assay achieved 100% sensitivity (71/71) and 100% specificity (42/42) compared to the qPCR. Conclusions In resource-limited settings, our method requires only a portable heat block or water bath and a blue light or ultraviolet flashlight for visual detection of MPXV, making it highly accessible. The integration of LAMP and CRISPR/Cas12b provides a robust, user-friendly platform for point-of-care testing, with promising potential for the rapid molecular diagnosis of infectious diseases. Supplementary Information The online version contains supplementary material available at 10.1186/s12985-025-02780-0.
研究领域:
医学
·
分子生物学
·
传染病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF MEDICAL VIROLOGY
·
2022
·Vol. 95
·期 1
·e28299
·
IF 20.693
Lu Lv,Leiliang Zhang
Monkeypox virus (MPXV) is a member of Orthopoxvirus in the Poxviridae family, causing a Public Health Emergency of International Concern. The number of cases and geographic range has increased significantly in 2022. Identification of MPXV-specific therapeutic targets is urgent. G-quadruplex (GQ) secondary structures attract great attention as potential targets for antiviral strategy. Whether GQs are present in the MPXV genome remains inconclusive. In this study, we aim to characterize the GQs encoded by MPXV. Through a series of biophysical experiments, we characterized the formation potential of MPXV-encoded GQs and evaluated the binding and stabilization abilities of GQ ligands including BRACO-19, pyridostatin, and TMPyP4 to GQs encoded by MPXV. Moreover, GQ ligands suppressed the gene transcription of MPXV sequences containing GQ. BRACO-19 and TMPyP4 were able to inhibit vaccinia virus replication. We demonstrated the existence of MPXV GQ and reinforced the idea that GQs could be novel antiviral targets. Targeting these GQ sequences with GQ-binding molecules may represent a new approach for MPXV therapy.
研究领域:
病毒学
·
分子生物学
·
药物开发
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Yu Tao,Haixia Wang,Enguo Ju,Yeh-Hsing Lao,Yu Zhang,Mingqiang Li
Diseases caused by viruses, such as monkeypox virus (MPXV) and human papillomavirus (HPV), pose serious threats to human health and safety. Although numerous strategies have been constructed for detecting MPXV and HPV DNA, most methods require either laborious procedures or complicated instruments involving skilled professionals. In this research, a CRISPR-Cas12a-mediated colorimetric detection platform for MPXV and HPV sensing was constructed for the first time by applying probe DNA to reprogram the catalytic properties of molybdenum disulfide quantum dots (MoS 2 QDs). In the presence of MPXV or HPV targets, the CRISPR-Cas12a trans -cleavage activity is effectively motivated to decompose the probe DNA, leading to the suppression of enzymatic activity DNA enhancer adsorbed on MoS 2 QDs, resulting in greatly decreased catalytic behaviors. The MoS 2 QDs-DNA nanohybrids displayed prominent specificity and sensitivity, with detection limits at subpicomolar levels, as well as excellent stability and accuracy for determining MPXV and HPV DNA in human sera biosamples. Furthermore, the proposed colorimetric biosensing approach can be ensembled with a smartphone platform, allowing visible analysis of DNA targets. Taken together, this colorimetric strategy offers a novel diagnosis method for MPXV and HPV DNA detection, particularly favorable for highly endemic developing countries with restricted medical and instrumental support.
研究领域:
分子生物学
·
病毒学
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Chen Chen,Yu Chen,Zhiyi Ye,Ahmad Ali,Shun Yao
To address the issues of infectious virus, bacterial secondary infections, skin pigmentation, and scarring caused by monkeypox virus (MPXV), a sprayable hydrogel with versatile functions was developed with comprehensive properties. Based on current research, the bioactive deep eutectic solvent (DES) of rosmarinic acid-proanthocyanidin-glycol (RPG) was designed and synthesized as active agent, and molecular docking was applied to discover its binding to MPXV proteins through H-bonds and van der Waals interactions, and the docking results show the binding energies between RA, PC, Gly and MPXV proteins are −58.7188, −50.2311, and −18.4755 kcal/mol, respectively. Additionally, poly(vinyl alcohol) (PVA), borate, and xylitol (Xyl) were integrated with RPG to prepare the PB-RPG-Xyl hydrogel, which was characterized by popular ways. The pH-responsive properties of the hydrogel accelerated the release of RPG under acidic conditions, resulting in an increased cumulative release percentage of 84.83% at pH 5.5 at 210 min. Besides that, it was proved to have the expected sprayability, self-healing, adhesion, and shape-adaptability. The results of molecular dynamic simulation were meaningful to understanding its formation and self-healing mechanisms. Furthermore, the hydrogel shows ideal degradability, removability, and biocompatibility. Lastly, its multiple functions were systematically explored, including UV-blocking, blood clotting, cooling, antioxidant, antibacterial, and virus inhibition properties. The developed sprayable PB-RPG-Xyl hydrogel represents the first promising dressing based on natural bioactive DES for MPXV lesions management, which not only expands the application of green solvents in health care but also provides a convenient and effective treatment process for MPXV infection in the face of difficult skin lesions and complex treatment needs.
研究领域:
材料科学
·
生物学
·
医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Runlin Yang,Siying Xie,Bin Zhou,Mingming Guo,Jun Fan,Fengli Su,Zhirun Ji,Yue Chen,Bingzhi Li
CRISPR/Cas12a-based biosensing is advancing rapidly; however, achieving sensitive and cost-effective reporting of Cas12a activation remains a challenge. In response, we have developed a label-free system capable of postamplifying Cas12a activation by integrating hybridization chain reaction (HCR) and DNA-copper nanoclusters (DNA-CuNCs). The trans-cleavage of Cas12a triggers a silenced HCR, leading to the in situ assembly of fluorescent DNA-CuNCs, allowing for the turn-on reporting of Cas12a activation. Without preamplification, this assay can detect DNA with a detection limit of 5 fM. Furthermore, when coupled with preamplification, the system achieves exceptional sensitivity, detecting the monkeypox virus (MPXV) plasmid at 1 copy in human serum. In a MPXV pseudovirus-based validation test, the obtained results are in agreement with those obtained by qPCR, reinforcing the robustness of this method. Our study represents the first effort to manipulate DNA-CuNC formation on HCR for highly sensitive and cost-effective reporting of Cas12a, resulting in an efficient synthetic biology-enabled sensing platform for biosafety applications.
研究领域:
合成生物学
·
生物传感
·
分子诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2023
·Vol. 15
·期 17
·2105-2113
·
IF 3.1
Furong Zhao,Pei Wang,Haoxuan Wang,Sirui Liu,Muhammad Sohail,Xing Zhang,Bingzhi Li,He Huang
The spread of the monkeypox virus (MPXV) from Central and West Africa to previously non-endemic regions has caused a global panic. In this context, the rapid, specific, and ultrasensitive detection of MPXV is crucial to contain its spread, though such technology has seldom been reported. Herein, we proposed an MPXV assay combining recombinase-aided amplification (RAA) and CRISPR/Cas12a. This assay targeted the highly conserved MPXV F3L gene and demonstrates a low detection limit (LOD) of 101 copies per μL. By leveraging the high specificity nature of RAA and CRISPR/Cas12a, we rationally optimized probes and conditions to achieve high selectivity that differentiates MPXV from other orthopox viruses and current high-profile viruses. To facilitate on-site screening of potential MPXV carriers, a kit integrating lateral flow strips was developed, enabling naked-eye MPXV detection with a LOD of 104 copies per μL. Our RAA-Cas12a-MPXV assay was able to detect MPXV without the need for sophisticated operation and expensive equipment. We believe that this assay can be rapidly deployed in emerging viral outbreaks for on-site surveillance of MPXV.
研究领域:
病毒学
·
分子诊断
·
生物技术
✓ 已验证
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Shenghao Zhang,Meiling Zhang,Yinhao Zhang,Yingxia Zong,Xu Hun
Here, we designed a catalytic DNA assembly (CDA) amplification strategy to construct a photoelectrochemical (PEC) assay to detect the specific target F3L-gene of Monkeypox Virus (MPXV). Graphdiyne (GDY) and CuSe were synthesized by a surface-assisted deprotection/polymerization method and hydrothermal method, respectively. Then AuNPs and GDY/CuSe nanocomposites were modified on the surface of gold electrode (GE). The hairpin DNA4 (H4) was incubated onto the surface of the electrode by Au-S bond. When the target was present, the CDA was occurred by hairpin DNA1 (H1) and hairpin DNA2 (H2). Then, DNA3 (H3) hybridized with the single stranded region of H1-H2 complex, formed a site that can be hydrolyzed by exonuclease III, led to exonuclease III hydrolysis of DNA and to release the H3 primer (H3P). H3 further opened H4 and triggered a subsequent CDA reaction, ultimately formed a Y-shaped DNA structure on the electrode surface with the help of DNA5 (H5) and DNA6 (H6). The two end sequences of H4 and H6, which constituted the Y-shaped structure, were designed as split H3P fragments. They can be used as primers to trigger CDA reactions for more efficient and characteristic autocatalytic signal amplification. The Y-shaped structure carried H5-ALP, and catalyzed phosphonoxyphenol ( o -PP) to generate benzene-1,2-diol (CA), which could enhance the PEC signal. The linear range is 0.1 fM-10.0 pM with the detection limit of 0.05 fM (3S/N) for MPXV.
研究领域:
生物医学
·
分子诊断
·
纳米材料科学
✓ 已验证
产品关联
DOI
高影响因子
Chemosensors
·
2025
·Vol. 13
·期 4
·142
·
IF 3.7
Wei-Zhi Zhang,Chen-Fei Zhang,Shou-Nian Ding
The Monkeypox virus (MPXV), a DNA virus classified under the Orthpoxvirus genus alongside variola virus, has recently garnered significant global health attention due to its increasing transmission and emerging genomic mutations. Point-of-care testing is essential for effective clinical response and outbreak mitigation. In this article, we developed a novel class of colored microspheres designed for application in a lateral flow immunoassay (LFIA) platform targeting MPXV-specific biomarkers. Polystyrene-maleic anhydride (SMA-MAA) microspheres were synthesized with a high-temperature soap-free emulsion polymerization optimized in our lab. Subsequent alkali and acid treatments were employed to introduce porosity into the microsphere matrix. Solvent Red 27 and Disperse Red 60 were incorporated via solvent-swelling and thermal-swelling methods, respectively, to generate high brightness (HB) carriers. A surface coating composed of a tannic acid–iron (TA–Fe3⁺) coordination complex was applied to form a stable metal–polyphenol film (MPF). This coating not only minimized dye leaching by establishing a robust shell but also improved dye distribution, thereby enhancing overall color intensity. The final HB-LFIA system, configured in a sandwich immunoassay format, demonstrated favorable sensitivity and linear detection range for Monkeypox antigen, indicating strong potential for clinical diagnostic use.
研究领域:
病毒学
·
医学诊断
·
材料科学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL LETTERS
·
2023
·
IF 2
Peipei Cao,Xiangde Lai,Rui Zhang,Zhiling Chen,Bin Qiao,Lina Niu,Qiang Wu
Monkeypox, a rare zoonotic disease classified as a category B infectious disease, is caused by the monkeypox virus (MPXV), which has a broad host range. Since May 2022, human MPXV infection outbreak has spread to more than 100 countries, and China has reported 106 MPXV infections between June 2 and 30, 2023. Therefore, rapid and effective MPXV detection is essential to control and prevent its transmission. In this study, a rapid, cost-effective, and convenient fluorescent immunochromatographic assay (FICA) for MPXV antigen A35R detection was developed. Under the optimal reaction conditions, the visual detection limit of the proposed assay was 7.5 ng/mL and was completed within 15 min. There was no cross-reactivity with norovirus (NV) or dengue virus (DENV) controls. Additionally, the clinical application of the proposed method was demonstrated by testing human serum samples spiked with the MPXV antigen A35R. FICA was faster and more convenient than traditional enzyme-linked immunosorbent assays (ELISA) or polymerase chain reaction (PCR), and hence suitable for the early screening of MPXV infection. In summary, FICA could realize rapid, low-cost, sensitive, and specific MPXV detection hold potential application prospects in clinical settings.
研究领域:
病毒学
·
传染病
·
分子诊断
·
临床微生物学
✓ 已验证
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Jia Wei,Wanjun Wang,Qiong Yu,Mingrui Zhang,Fengyu Xue,Beibei Fan,Tong Zhang,Yanpeng Gao,Juan Li,Xianying Meng,Bo Pang
Mpox (formerly referred as Monkeypox) outbreak has been declared a Public Health Emergency of International Concern. However, traditional polymerase chain reaction (PCR) diagnostic technology is not ideal for on-site applications. To conduct the sample-to-result Mpox viral particles detection outside the laboratories, we developed an easy-to-operate palm-size pouch, termed Mpox At-home Self-Test and point-of-caRe Pouch (MASTR Pouch). In this MASTR Pouch, the fast and accurate visualization was achieved by incorporating recombinase polymerase amplification (RPA) with clustered regularly interspaced short palindromic repeat (CRISPR)/Cas12a system. From viral particle lysis to naked eye readout, MASTR Pouch required only four simple steps to accomplish the analysis process within 35 min. Fifty-three Mpox pseudo-viral particles in exudate (10.6 particles/μL) were able to be detected. To verify the practicability, 104 mock Mpox clinical exudate specimens were tested. The clinical sensitivities were determined to be 91.7%− 95.8%. There was no false-positive result, validating the 100% clinical specificity. MASTR Pouch approaches the WHO's ASSURD criteria for point-of-care diagnostic, which will be beneficial for mitigating Mpox's global spread. The versatility potential of MASTR Pouch could further revolutionize infection diagnosis.
研究领域:
病毒学
·
分子诊断
·
公共卫生
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Lunjing Liu,Yu Fu,Chengli Zhang,Jingyan Lin,Huiyi Yang,Wenjie Wu,Jingru Wang,Jiang Xiao,Yideng Jiang,Yuhui Liao,Guanghua Li
Recurrent outbreaks of Mpox have highlighted the importance of rapid diagnosis to effectively control virus transmission from infected individuals. Traditional laboratory-based detection methods are time-consuming, complex, and unsuitable for point-of-care testing. Therefore, we developed a lateral flow immunoassay (LFIA) using platinum–gold nanoflowers (PtAu NFs) as a labeling material, and this technique was integrated with recombinase polymerase amplification (RPA) to detect Mpox within 25 min. From material preparation to detection, the speed and simplicity of detection were comprehensively optimized in this study. Moreover, a novel material preparation method based on tripropylamine (TPrA) as a reducing agent for the rapid preparation of PtAu NFs has been demonstrated. Unlike traditional reducing agents, this method does not require high temperatures and extended incubation periods, thereby providing a novel strategy for preparing nanomaterials. Furthermore, the traditional RPA–LFIA strategy was improved by directly modifying the primer label, avoiding the use of probes and exonucleases, which saves reagent costs. Under optimized conditions, the RPA–PtAu–LFIA platform had a visual limit of detection of 1 copy/μL, with high specificity for Mpox and other poxviruses. Furthermore, the sensitivity of this platform was validated using 29 different clinical samples obtained from the pustules, saliva, anus, and throat of infected individuals; the results obtained using this platform were comparable to those obtained using quantitative polymerase chain reaction. This study provides an excellent platform for preparing nanomaterials and detecting Mpox.
研究领域:
病毒学
·
纳米医学
·
诊断学
✓ 已验证
产品关联
DOI
高影响因子
Emerging Microbes & Infections
·
2024
·
IF 8.4
Anke Chen,Ning Fang,Zhifei Zhang,Yiqing Wen,Yajie Shen,Yanjie Zhang,Lu Zhang,Guoping Zhao,Jin Ding,Jixi Li
The global outbreak of Mpox, caused by the monkeypox virus (MPXV), has attracted international attention and become another major infectious disease event after COVID-19. The mRNA cap N7 methyltransferase (RNMT) of MPXV methylates the N7 position of the added guanosine to the 5'-cap structure of mRNAs and plays a vital role in evading host antiviral immunity. MPXV RNMT is composed of the large subunit E1 and the small subunit E12. How E1 and E12 of MPXV assembly remains unclear. Here, we report the crystal structures of E12, the MTase domain of E1 with E12 (E1CTD-E12) complex, and the E1CTD-E12-SAM ternary complex, revealing the detailed conformations of critical residues and the structural changes upon E12 binding to E1. Functional studies suggest that E1CTD N-terminal extension (Asp545-Arg562) and the small subunit E12 play an essential role in the binding process of SAM. Structural comparison of the AlphaFold2-predicted E1, E1CTD-E12 complex, and the homologous D1-D12 complex of vaccinia virus (VACV) indicates an allosteric activating effect of E1 in MPXV. Our findings provide the structural basis for the MTase activity stimulation of the E1-E12 complex and suggest a potential interface for screening the anti-poxvirus inhibitors.
研究领域:
病毒学
·
结构生物学
·
分子生物学
·
传染病学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）