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A global outbreak of monkeypox (mpox) caused by the mpox virus (MPXV) has posed a serious threat to public health worldwide, thus calling for the urgent development of antivirals and vaccines to curb its further spread. In this study, we screened 41 anhydride-modified proteins and found that 3-hydroxyphthalic anhydride-modified ß-lactoglobulin (3HP-ß-LG), a clinically used anti-HPV agent, was highly effective in inhibiting infection of vaccinia virus Tiantan strain (VACV-VTT) and MPXV. Mechanistic studies demonstrated that 3HP-ß-LG bound to the virus, not the host cell, by targeting the early stage of virus entry, possibly through the interaction between the amino acids with negatively charges in 3HP-ß-LG and the key amino acids with positive charges in the target region of A29L, a key surface protein of MPXV. A synergistic effect was observed when 3HP-ß-LG was combined with tecovirimat, a small-molecule antiviral drug approved by the United States Food and Drug Administration and the European Medicine Agency for the treatment of smallpox and mpox. Because of its clinically proven safety and stability, 3HP-ß-LG shows promise for further development as a prophylactic agent to prevent the sexual transmission of MPXV.
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Abnormal trophoblast self-renewal and differentiation during early gestation is the major cause of miscarriage, yet the underlying regulatory mechanisms remain elusive. Here, we show that trophoblast specific deletion of Kat8, a MYST family histone acetyltransferase, leads to extraembryonic ectoderm abnormalities and embryonic lethality. Employing RNA-seq and CUT&Tag analyses on trophoblast stem cells (TSCs), we further discover that KAT8 regulates the transcriptional activation of the trophoblast stemness marker, CDX2, via acetylating H4K16. Remarkably, CDX2 overexpression partially rescues the defects arising from Kat8 knockout. Moreover, increasing H4K16ac via using deacetylase SIRT1 inhibitor, EX527, restores CDX2 levels and promoted placental development. Clinical analysis shows reduced KAT8, CDX2 and H4K16ac expression are associated with recurrent pregnancy loss (RPL). Trophoblast organoids derived from these patients exhibit impaired TSC self-renewal and growth, which are significantly ameliorated with EX527 treatment. These findings suggest the therapeutic potential of targeting the KAT8-H4K16ac-CDX2 axis for mitigating RPL, shedding light on early gestational abnormalities.
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Fator de Transcrição CDX2 , Proliferação de Células , Autorrenovação Celular , Histona Acetiltransferases , Trofoblastos , Trofoblastos/metabolismo , Fator de Transcrição CDX2/metabolismo , Fator de Transcrição CDX2/genética , Animais , Feminino , Humanos , Camundongos , Gravidez , Autorrenovação Celular/genética , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/genética , Aborto Habitual/metabolismo , Aborto Habitual/genética , Camundongos Knockout , Histonas/metabolismo , Diferenciação Celular , Placentação/genéticaRESUMO
Since May 2022, the multi-country outbreak of monkeypox (mpox) has raised a great concern worldwide. Early detection of mpox virus infection is recognized as an efficient way to prevent mpox transmission. Mpox specific detection methods reported up to now are based on the SNPs among mpox virus and other orthopoxviruses. We have therefore developed a real-time PCR based mpox detection method targeting mpox virus specific sequences (N3R and B18Rplus). We have also optimized an orthopoxvirus detection system which targets the highly conserved E9L and D6R genes. The mpox and orthopoxvirus real-time PCR assays have a high sensitivity (1 copy/reaction) and specificity. Mpox viral DNA and clinical samples from mpox patients are detected with the mpox detection system. Furthermore, we have established a multiplex real-time PCR detection system allowing simultaneous and efficient detection of mpox and orthopoxvirus infections.
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Monkeypox virus , Mpox , Reação em Cadeia da Polimerase Multiplex , Orthopoxvirus , Infecções por Poxviridae , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Orthopoxvirus/genética , Orthopoxvirus/isolamento & purificação , Humanos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Monkeypox virus/genética , Monkeypox virus/isolamento & purificação , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/virologia , Infecções por Poxviridae/veterinária , Mpox/diagnóstico , Mpox/virologia , Técnicas de Diagnóstico Molecular/métodosRESUMO
The increasing global popularity of ready-to-eat (RTE) foods for their convenience simultaneously brings along a risk, as these products can be contaminated with various microorganisms, including potentially harmful pathogens. We aimed to investigate the food contamination of Staphylococcus aureus (S. aureus) in RTE foods in Guangdong, South China. All S. aureus isolates were subjected to characterization through antimicrobial susceptibility tests, multilocus sequence typing (MLST), and PCR analysis for detecting mec and blaZ genes. A total of 824 RTE food samples were collected from 2017 to 2022, of which 73 (8.9%) were found to be contaminated with S. aureus. Contamination levels were mostly in the range of 0.3-1.0 most probable number (MPN)/g, with 10 samples exceeding 110 MPN/g. Of the 73 S. aureus isolates, 10 were identified as methicillin-resistant S. aureus (MRSA). In MRSA, resistance was most frequently observed to penicillin (100%, 10/10), followed by erythromycin (80.0%, 8/10) and tetracycline (70%, 7/10). And in methicillin-sensitive S. aureus (MSSA), resistance was most frequently observed to penicillin (98.4%, 62/63), followed by tetracycline (30.2%, 19/63) and erythromycin (23.8%, 15/63). Overall, 98.6% (72/73) of the isolates demonstrated resistance to at least one antimicrobial agent, whereas 31.5% (23/73) were resistant to three or more antimicrobials. Fifty-seven S. aureus isolates harbored the penicillin-resistant gene blaZ, and 10 isolates carried the mec gene. In addition, 30.1% of the isolates harbored genes for classical staphylococcal enterotoxins (SEs), with seb being the most frequently detected SE gene. MLST revealed that the 73 isolates belonged to 14 different sequence types (STs), the most prevalent of which was ST7. In MRSA, the most common prevalent clone is ST6, and in MSSA, ST7 was the most common isolates. The prevalent multidrug resistance indicates that the resistance situation of foodborne S. aureus in Guangdong is severe, posing a potential threat to consumer safety and health.
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The monkeypox virus (MPXV) has triggered a current outbreak globally. Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control. It is a significant challenge but necessary to optimize the strategy and application of rapid full-length genome identification and to track variations of MPXV in clinical specimens with low viral loads, as it is one of the DNA viruses with the largest genome and the most AT-biased, and has a significant number of tandem repeats. Here we evaluated the performance of metagenomic and amplicon sequencing techniques, and three sequencing platforms in MPXV genome sequencing based on multiple clinical specimens of five mpox cases in Chinese mainland. We rapidly identified the full-length genome of MPXV with the assembly of accurate tandem repeats in multiple clinical specimens. Amplicon sequencing enables cost-effective and rapid sequencing of clinical specimens to obtain high-quality MPXV genomes. Third-generation sequencing facilitates the assembly of the terminal tandem repeat regions in the monkeypox virus genome and corrects a common misassembly in published sequences. Besides, several intra-host single nucleotide variations were identified in the first imported mpox case. This study offers an evaluation of various strategies aimed at identifying the complete genome of MPXV in clinical specimens. The findings of this study will significantly enhance the surveillance of MPXV.
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Monkeypox virus , Mpox , Humanos , Monkeypox virus/genética , Mpox/diagnósticoRESUMO
Investigating immune memory to vaccinia virus and pre-existing immunity to mpox virus (MPXV) among the population is crucial for the global response to this ongoing mpox epidemic. Blood was sampled from vaccinees inoculated with vaccinia virus Tiantan (VTT) strain born before 1981 and unvaccinated control subjects born since 1982. After at least 40 years of the inoculation, 60% or 5% VTT vaccinees possess neutralizing antibodies (NAbs) to VTT or MPXV, with at least 50% having T cell memory to VTT protein antigens. Notably, 46.7% vaccinees show pre-existing T cell responses to MPXV. Broad pre-existing CD8+ T cell reactivities to MPXV are detected not only against conserved epitopes but also against variant epitopes between VTT and MPXV. Persistent NAbs and T cell memory to VTT among vaccinees, along with pre-existing T cells to MPXV among both vaccinees and the unvaccinated population, indicate a particular immune barrier to mpox.
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Mpox , Vaccinia virus , Humanos , Monkeypox virus , Imunidade Celular , Anticorpos Neutralizantes , China , Epitopos , Imunidade HumoralRESUMO
BACKGROUND: It is important to extend the indication of severe acute respiratory syndrome coronavirus 2 vaccine to children to improve the vaccine intake rate and reduce infection in this population. METHODS: In 2 phase 1 and phase 2 randomized, double-blind and placebo-controlled trials, 84 and 480 Chinese healthy children 3 to 17 years old were enrolled, respectively, and randomized in 3:1 ratio to receive 2 doses of a severe acute respiratory syndrome coronavirus 2 inactivated vaccine, KCONVAC or placebo. The 2 doses were given 28 days apart. Adverse events (AEs) were recorded through Day 28 after each dosing. Live virus neutralizing antibody and receptor binding domain antibody (RBD-IgG) were tested before vaccination and after the second dose. RESULTS: Two doses of the vaccine, KCONVAC, elicited geometric mean titers of 142-150 for neutralizing antibody and 4154-4253 for RBD-IgG 28 days after the second dose. Seroconversion rates were 100% after 2 doses for both antibodies in both trials. The predominant AEs were injection-site pain, cough and fever. Most AEs were grade 1 or 2 in intensity. Five participants reported 6 vaccination-unrelated serious AEs in the phase 2 trial. CONCLUSIONS: Two doses of this study vaccine, KCONVAC, were well tolerated and immunogenic in children 3 to 17 years of age.
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Vacinas contra COVID-19 , COVID-19 , Imunogenicidade da Vacina , Adolescente , Criança , Pré-Escolar , Humanos , Anticorpos Neutralizantes , Anticorpos Antivirais , COVID-19/prevenção & controle , Método Duplo-Cego , População do Leste Asiático , Imunoglobulina G , SARS-CoV-2 , Vacinas de Produtos Inativados/efeitos adversos , Vacinas de Produtos Inativados/imunologia , Vacinas contra COVID-19/efeitos adversos , Vacinas contra COVID-19/imunologiaRESUMO
Mpox is caused by a zoonotic virus belonging to the Orthopoxvirus genus and the Poxviridae family. In this study, we develop a recombinase polymerase amplification (RPA)-coupled CRISPR-Cas12a detection assay for the mpox virus. We design and test a series of CRISPR-derived RNAs(crRNAs) targeting the conserved D6R and E9L genes for orthopoxvirus and the unique N3R and N4R genes for mpox viruses. D6R crRNA-1 exhibits the most robust activity in detecting orthopoxviruses, and N4R crRNA-2 is able to distinguish the mpox virus from other orthopoxviruses. The Cas12a/crRNA assay alone presents a detection limit of 108 copies of viral DNA, whereas coupling RPA increases the detection limit to 1-10 copies. The one-tube RPA-Cas12a assay can, therefore, detect viral DNA as low as 1 copy within 30 min and holds the promise of providing point-of-care detection for mpox viral infection.
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Mpox , Orthopoxvirus , Humanos , Recombinases/genética , Sistemas CRISPR-Cas/genética , Monkeypox virus , DNA Viral/genética , Nucleotidiltransferases , RNA Guia de Sistemas CRISPR-CasRESUMO
Current COVID-19 vaccines can effectively reduce disease severity and hospitalisation; however, they are not considerably effective in preventing infection and transmission. In this context, mucosal vaccines are pertinent to prevent SARS-CoV-2 infection and spread. In this study, we generated a replication-competent recombinant chimeric influenza A virus (IAV) expressing the receptor-binding domain (RBD) of a SARS-CoV-2 prototype in the C-terminus of the neuraminidase (NA) of A/Puerto Rico/08/1934 H1N1 (PR8). The remaining seven segments from A/WSN/1933 H1N1 (WSN) were named PR8NARBD/WSN. We observed that the recombinant virus with the WSN backbone demonstrated improved expression of NA and RBD. A single intranasal dose of PR8NARBD/WSN(103PFU) in mice generated robust mucosal immunity, neutralising antibodies, cellular immunity, and tissue-resident memory T cells specific to SARS-CoV-2 and IAV. Importantly, immunisation with PR8NARBD/WSN viruses effectively protected mice against lethal challenges with H1N1, H3N2 IAV, and SARS-CoV-2 Beta variant and significantly reduced lung viral loads. Overall, our research demonstrates the promising potential of PR8NARBD/WSN as an attractive vaccine against emerging SARS-CoV-2 variants and influenza A virus infections.
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Enveloped RNA viruses are a group of viruses with an outer membrane derived from a host cell and a genome consisting of ribonucleic acid (RNA). These viruses rely on host cell machinery and organelles to replicate and assemble new virus particles. However, the interaction between viruses and host organelles may be disrupted by nanomaterials, such as gold nanoparticles (AuNPs) with unique physical and chemical properties. In this study, we investigated the effects of AuNPs with different surface charge properties on the subcellular structure and function of mammalian cells, and their effects on two representative enveloped RNA viruses: lentivirus and human coronavirus OC43 (HCoV- OC43) antiviral potential. By comparing the subcellular effects of AuNPs with different surface charge properties, we found that treatment with AuNPs with positive surface charges induced more significant disruption of subcellular structures than neutrally charged AuNPs and negatively charged AuNPs, mainly manifested in lysosomes and Cytoskeletal disorders. The antiviral effect of the surface positively charged AuNPs was further evaluated using lentivirus and HCoV-OC43. The results showed that AuNPs had a significant inhibitory effect on both lentivirus and HCoV-OC43 without obvious side effects. In conclusion, our study provides insights into the mechanism of action and biocompatibility of AuNP in biological systems, while supporting the potential of targeting organelle dynamics against enveloped RNA viruses.
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Nanopartículas Metálicas , Vírus de RNA , Animais , Humanos , Ouro/farmacologia , Ouro/química , Ouro/metabolismo , Nanopartículas Metálicas/química , Organelas/metabolismo , Vírus de RNA/genética , MamíferosRESUMO
Many of the currently available COVID-19 vaccines and therapeutics are not effective against newly emerged SARS-CoV-2 variants. Here, we developed the metallo-enzyme domain of angiotensin converting enzyme 2 (ACE2)-the cellular receptor of SARS-CoV-2-into an IgM-like inhalable molecule (HH-120). HH-120 binds to the SARS-CoV-2 Spike (S) protein with high avidity and confers potent and broad-spectrum neutralization activity against all known SARS-CoV-2 variants of concern. HH-120 was developed as an inhaled formulation that achieves appropriate aerodynamic properties for rodent and monkey respiratory system delivery, and we found that early administration of HH-120 by aerosol inhalation significantly reduced viral loads and lung pathology scores in male golden Syrian hamsters infected by the SARS-CoV-2 ancestral strain (GDPCC-nCoV27) and the Delta variant. Our study presents a meaningful advancement in the inhalation delivery of large biologics like HH-120 (molecular weight (MW) ~ 1000 kDa) and demonstrates that HH-120 can serve as an efficacious, safe, and convenient agent against SARS-CoV-2 variants. Finally, given the known role of ACE2 in viral reception, it is conceivable that HH-120 has the potential to be efficacious against additional emergent coronaviruses.
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Enzima de Conversão de Angiotensina 2 , COVID-19 , Masculino , Animais , Cricetinae , Humanos , Vacinas contra COVID-19 , SARS-CoV-2/genética , Mesocricetus , Imunoglobulina MRESUMO
BACKGROUND: Several COVID-19 vaccines are in widespread use in China. Few data exist on comparative immunogenicity of different COVID-19 vaccines given as booster doses. We aimed to assess neutralizing antibody levels raised by injectable and inhaled aerosolized recombinant adenovirus type 5 (Ad5)-vectored COVID-19 vaccine as a heterologous booster after an inactivated COVID-19 vaccine two-dose primary series. METHODS: Using an open-label prospective cohort design, we recruited 136 individuals who had received inactivated vaccine primary series followed by either injectable or inhaled Ad5-vectored vaccine and measured neutralizing antibody titers against ancestral SARS-CoV-2 virus and Omicron BA.1 and BA.5 variants. We also measured neutralizing antibody levels in convalescent sera from 39 patients who recovered from Omicron BA.2 infection. RESULTS: Six months after primary series vaccination, neutralizing immunity against ancestral SARS-CoV-2 was low and neutralizing immunity against Omicron (B.1.1.529) was lower. Boosting with Ad5-vectored vaccines induced a high immune response against ancestral SARS-CoV-2. Neutralizing responses against Omicron BA.5 were ≥ 80% lower than against ancestral SARS-CoV-2 in sera from prime-boost subjects and in convalescent sera from survivors of Omicron BA.2 infection. Inhaled aerosolized Ad5-vectored vaccine was associated with greater neutralizing titers than injectable Ad5-vectored vaccine against ancestral and Omicron SARS-CoV-2 variants. CONCLUSIONS: These findings support the current strategy of heterologous boosting with injectable or inhaled Ad5-vectored SARS-CoV-2 vaccination of individuals primed with inactivated COVID-19 vaccine.
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Vacinas contra COVID-19 , COVID-19 , Humanos , Anticorpos Neutralizantes , Anticorpos Antivirais , Estudos Prospectivos , SARS-CoV-2RESUMO
Introduction: Since 2019, numerous variants of concern for severe acute respiratory syndrome virus 2 (SARS-CoV-2) have emerged, leading to significant outbreaks. The development of novel, highly accurate, and rapid detection techniques for these new SARS-CoV-2 variants remains a primary focus in the ongoing efforts to control and prevent the coronavirus disease 2019 (COVID-19) pandemic. Methods: Reverse transcription-recombinase polymerase amplification combined with the clustered regularly interspaced short palindromic repeats-associated protein 12a (CRISPR/Cas12a) system was used to validate the detection of the Omicron BA.2, BA.4, and BA.5 variants of SARS-CoV-2. Results: Our results demonstrate that the CRISPR/Cas12a assay is capable of effectively detecting the SARS-CoV-2 BA.2, BA.4, and BA.5 variants with a limit of detection of 10, 1, and 10 copies/µL, respectively. Importantly, our assay successfully differentiated the three SARS-CoV-2 Omicron strains from one another. Additionally, we evaluated 46 SARS-CoV-2 positive clinical samples consisting of BA.2 (n=20), BA.4 (n=6), and BA.5 (n=20) variants, and the sensitivity of our assay ranged from 90% to 100%, while the specificity was 100%. Discussion: This research presents a swift and reliable CRISPR-based method that may be employed to track the emergence of novel SARS-CoV-2 variants.
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BACKGROUND AND OBJECTIVE: Kawasaki disease (KD) is an acute self-limiting systemic vascular disease commonly observed in children less than 5 years of age. The present study comparatively assesses the clinical characteristics of children diagnosed with KD in different age groups. Furthermore, a comprehensive literature review on the clinical features and diagnostic guidelines of KD is performed. METHODS: This was a retrospective study conducted on the data of KD children admitted to the Sun Yat-Sen Memorial Hospital, Guangzhou, China, from January 2016 to December 2018. The children were divided into 3 age groups, including children < 1 year of age (group A, n = 66), 1-5 years of age (group B, n = 74), and children > 5 years of age (group C, n = 14). Complete clinical evaluation, hematological, and cardiovascular assessments were conducted and compared between the three groups. RESULTS: The time of diagnosis, hemoglobin, and neutrophil ratio of children in group A were significantly lower than the other two groups (p < 0.05), while the platelet count was significantly higher (p < 0.05). The proportion of incomplete KD (iKD) was the greatest in group A (40.9%), while the proportion of children with increased coronary Z value and aseptic meningitis was greater than that in group B (p < 0.0167). Group A showed less patients with KD shock syndrome (KDSS) than the other two groups (p < 0.05). Group B showed the greatest number of patients with arthralgia compared to the other two groups (p < 0.05). Three groups showed no significant difference to intravenous immunoglobulin (IVIG) therapy (p > 0.05). CONCLUSION: The younger the age of KD onset, the more atypical the conditions are, with a greater risk of affecting other systems and a higher incidences of coronary artery disease. An early treatment with glucocorticoids might be helpful in older children and those with a greater high-risk KD warning score to prevent coronary injury.
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Doença da Artéria Coronariana , Síndrome de Linfonodos Mucocutâneos , Choque , Criança , Humanos , Lactente , Estudos Retrospectivos , Síndrome de Linfonodos Mucocutâneos/diagnóstico , Síndrome de Linfonodos Mucocutâneos/tratamento farmacológico , Síndrome de Linfonodos Mucocutâneos/epidemiologia , Imunoglobulinas Intravenosas/uso terapêutico , Doença da Artéria Coronariana/tratamento farmacológico , Choque/tratamento farmacológicoRESUMO
Substantial evidence links physical activity and sedentary behaviour (SB) with cardiovascular health. However, studies usually examine time-use behaviours independently, rather than considering daily activities as a 24-hour time-use composition. This study aimed to use compositional data models to investigate how time reallocations from SB to moderate-to-vigorous physical activity (MVPA)/ light physical activity (LPA) were associated with changes in cardiometabolic biomarkers. Compositional isotemporal substitution analysis was conducted using data from a representative cross-sectional sample of adults aged 20 years or older (n = 5213 full sample; n = 2221 fasting subsample) from the 2003-2004 and 2005-2006 cycles of the National Health and Nutrition Examination Survey. Overall as well as gender- and age-specific estimates were obtained for reallocating time between the behaviours associated with cardiometabolic outcomes. Replacing SB with MVPA was associated with decreased levels of C-reactive protein (CRP), white blood cells (WBCs), red blood cell distribution width (RDW), insulin, insulin resistance homeostasis model assessment (HOMA-IR) and 2-hour glucose, and increased high-density lipoprotein cholesterol (HDL-C), except for WBCs in older adults, insulin and HOMA-IR in females and older adults, 2-hour glucose in young and older adults, and HDL-C in older adults. Substituting LPA for SB resulted in lower CRP in older adults, lower RDW in males and older adults, lower insulin in young and middle-aged adults, lower HOMA-IR in females and middle-aged adults, and higher HDL-C in males and young adults. Our findings provide new evidence for the potential benefits of LPA and MVPA in cardiovascular health and help make population-specific recommendations for physical activity.HighlightsShorter sedentary behaviour (SB) time and longer active behaviours time are associated with a lower cardiovascular disease risk profile.Although replacing SB with MVPA has stronger positive health effects, LPA, as a simpler and more feasible activity, may has greater potential in promoting cardiovascular health.The expected changes in cardiometabolic biomarkers caused by time reallocations from SB to MVPA/LPA vary with gender and age, and physical activity guidelines with specific recommendations should be tailed to the subpopulations.
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Doenças Cardiovasculares , Comportamento Sedentário , Masculino , Pessoa de Meia-Idade , Feminino , Adulto Jovem , Humanos , Idoso , Estudos Transversais , Inquéritos Nutricionais , Acelerometria/métodos , Exercício Físico , Biomarcadores , Proteína C-Reativa , Insulina , GlucoseRESUMO
The emergence of new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has generated recurring worldwide infection outbreaks. These highly mutated variants reduce the effectiveness of current coronavirus disease 2019 (COVID-19) vaccines, which are designed to target only the spike (S) protein of the original virus. Except for the S of SARS-CoV-2, the immunoprotective potential of other structural proteins (nucleocapsid, N; envelope, E; membrane, M) as vaccine target antigens is still unclear and worthy of investigation. In this study, synthetic DNA vaccines encoding four SARS-CoV-2 structural proteins (pS, pN, pE, and pM) were developed, and mice were immunized with three doses via intramuscular injection and electroporation. Notably, co-immunization with two DNA vaccines that expressed the S and N proteins induced higher neutralizing antibodies and was more effective in reducing the SARS-CoV-2 viral load than the S protein alone in mice. In addition, pS co-immunization with either pN or pE + pM induced a higher S protein-specific cellular immunity after three immunizations and caused milder histopathological changes than pS alone post-challenge. The role of the conserved structural proteins of SARS-CoV-2, including the N/E/M proteins, should be investigated further for their applications in vaccine design, such as mRNA vaccines.
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Introduction: The first imported case of monkeypox (MPX) from the mainland of China was reported in September 2022. Herein, the study reports the isolation and characterization of MPX virus (MPXV) in this case. Methods: Clinical specimens including skin blister fluid, oropharyngeal and nasopharyngeal swabs, and blood were collected and inoculated onto Vero cells. The isolated virus was identified as MPXV using quantitative polymerase chain reaction (qPCR), cytopathic effects (CPEs), immunofluorescence assay (IFA) and transmission electron microscopy (TEM). Plaque assays were employed to quantify infectious plaque-forming units (PFUs). The plaque reduction neutralization test (PRNT) was developed to determine the neutralizing antibody (nAb) against MPXV. Results: MPXV replication was confirmed with qPCR. Typical CPEs were observed 48 h post-incubation. The isolated virus was named MPXV-B.1-China-C-Tan-CQ01. IFA showed that MPXV reacted with serum of MPX case. Orthopoxvirus morphology was observed using TEM. The virus titer increased to >106 PFU/mL after three passages. The serum PRNT 50% neutralization titer (NT50) was 35 for the MPX patient 6 days after symptom onset. Discussion: The study successfully isolated the first MPXV strain in the mainland of China, MPXV-B.1-China-C-Tan-CQ01. Infectious titration and PRNT methods have been developed. The study provides key resources and technical platforms for further research as well as anti-viral drug and vaccine development against MPX.