Introduction of monkeypox virus in Benin,2022

Dear Editor,Monkeypox is an infectious disease that is endemic in a dozen of African countries.Some imported cases have been also reported outside of Africa in the past[1].Since early May 2022,monkeypox infections including human-to-human transmission,were reported in a multi-country outbreak in non-endemic countries and declared Public Health Emergency of International Concern(PHEIC)by the World Health Organization(WHO)in July 2022[2].As of 20 September 2022,a total of at least 62,798 human cases of monkeypox with 20 deaths have been confirmed in 115 countries in five WHO regions[3].

An unfolding monkeypox outbreak in Europe and beyond

Dear Editor,Monkeypox is a zoonotic disease caused by the monkeypox virus(MPXV),a double-stranded DNA virus in the genus Orthopoxvirus,family Poxviridae.MPXV is known to be transmitted between animals such as rats and squirrels,but animal-to-human and human-to-human transmission has been known in Africa since 1970.Transmission is linked to direct contact with body fluids,skin lesions,and patient items as well as respiratory droplets in case of prolonged face-to-face contact[1].Until recently,only a few cases were associated with travel to endemic countries or contact with infected animals imported from endemic countries.

Emergency department approach to monkeypox

BACKGROUND:Monkeypox(mpox) is a viral infection that is primarily endemic to countries in Africa,but large outbreaks outside of Africa have been historically rare.In June 2022,mpox began to spread across Europe and North America,causing the World Health Organization(WHO) to declare mpox a public health emergency of international concern.This article aims to review clinical presentation,diagnosis,and prevention and treatment strategies on mpox,providing the basic knowledge for prevention and control for emergency providers.METHODS:We conducted a review of the literature using PubMed and SCOPUS databases from their beginnings to the end of July 2023.The inclusion criteria were studies on adult patients focusing on emerging infections that described an approach to a public health emergency of international concern,systematic reviews,clinical guidelines,and retrospective studies.Studies that were not published in English were excluded.RESULTS:We included 50 studies in this review.The initial symptoms of mpox are non-specific:fever,malaise,myalgias,and sore throat.Rash,a common presentation of mpox,usually occurs 2–4 weeks after the prodrome,but the presence of lymphadenopathy may distinguish mpox from other infections from the Poxviridae family.Life-threatening complications such as pneumonia,sepsis,encephalitis,myocarditis,and death can occur.There are documented co-occurrences of human immunodeficiency virus(HIV) and other sexually transmitted infections that can worsen morbidity.CONCLUSION:The initial presentation of mpox is non-specific.The preferred treatment included tecovirimat in patients with severe illness or at high risk of developing severe disease and vaccination with two doses of JYNNEOS.However,careful history and physical examination can raise the clinicians’ suspicion and point toward a prompt diagnosis.There are diff erent modalities to prevent and treat mpox infection.

Monkeypox outbreak in the age of COVID‑19:a new global health emergency

Dear Editor,The World Health Organization recently declared monkeypox as a global emergency after reporting more than 57,000 new cases worldwide[1].Monkeypox is an orthopoxvirus similar in nature to the Variola virus,which is a causative agent for smallpox[2].Monkeypox virus presents as a unique challenge for global health and should be regarded with grave concern as the vast majority of cases are occurring in countries where the disease is not considered endemic.Furthermore,the spread of this pathogen is occurring concomitantly as the world is still engaged in a battle with the coronavirus disease 2019(COVID-19)pandemic,which has caused substantial damage to global healthcare infrastructure.If monkeypox continues its rapid spread,hospitals could be quickly overwhelmed by both COVID-19 and monkeypox cases.Now more than ever,early symptom recognition and use of all available treatments to contain monkeypox outbreaks are vital.Early precautions including early recognition of symptoms by physicians,use of currently available treatments,and promoting precautionary measures in at-risk populations may be vital in preventing hospital burden and further physician burnout.

Near obstructing painful anorectal mass and facial rash in a man with monkeypox: A case report

BACKGROUND Monkeypox(MPX)is a zoonotic infection that is endemic in Western and Central Africa along the Congo River basin.It has a high case fatality rate especially in younger age groups.It belongs to the virus family orthopoxvirus like smallpox.It is transmitted from wild animals to humans but human to human transmission has been established.It is often a self-limited infection in endemic regions.Recently,attention has been given to MPX with the spread of infection to Europe and the United States of America(USA).There is currently sporadic infection of MPX in the USA especially amongst men who have sex with men(MSM).It is a serious life-threatening infection in human immunodeficiency virus/acquired immunodeficiency syndrome co-infected individuals especially those who are treatment naïve with severe immunosuppression.CASE SUMMARY We report a 38-year old man who presented with rectal pain,and anal,torso,and facial rash.Abdominal computed tomography scan showed a near obstructive rectal mass with peri-anal fistula.MPX was positive.He was started on tecovirimat(TPOXX)and HAART therapy.Additional treatment provided included vaccinia immunoglobulin following his clinical deterioration.CONCLUSION This case highlights a rare presentation of MPX with peri-anal fistula and near obstructive rectal mass,and the significance of MPX as a differential diagnosis in proctitis in MSM in addition to other sexually transmitted infection like gonorrhea and chlamydia.

猴痘病毒感染的神经系统表现研究进展

在过去的一年内,猴痘的爆发已成为全球关注的问题。猴痘是猴痘病毒感染引起的人畜共患病,除典型的皮疹症状以外,猴痘病毒感染还可引起一系列神经系统表现,潜在的机制可能包括感染后免疫介导的神经系统损伤,以及病毒直接侵入神经系统。本文围绕猴痘病毒感染的神经系统表现进行综述,以促进早期识别、诊断猴痘病毒感染神经系统并发症,及时采取相应的防治措施。

猴痘病毒A5L蛋白的原核表达及其免疫原性评价

【目的】获取高纯度猴痘病毒(Monkeypox virus, MPXV)A5L蛋白,制备小鼠抗A5L高效价抗血清,为MPXV A5L蛋白功能研究及相关诊断试剂的研发提供材料。【方法】构建重组质粒pET-28a-A5L,转化大肠杆菌BL21(DE3)感受态细胞进行诱导表达,通过分析不同IPTG诱导浓度、诱导温度、诱导时间条件下重组蛋白A5L表达情况,筛选最佳表达条件。通过Western blotting鉴定重组蛋白A5L表达,利用SDS-PAGE分析重组蛋白可溶性。A5L蛋白经His-Bind镍柱亲和层析纯化后免疫小鼠,通过ELISA法检测抗体效价。【结果】双酶切和测序结果显示,原核表达载体pET-28a-A5L构建成功。Western blotting结果显示重组蛋白A5L在大肠杆菌BL21(DE3)感受态细胞中成功表达;A5L蛋白在诱导温度42℃、诱导剂(IPTG)浓度为1.6 mmol/L、诱导16 h后表达量最高,大小为40 ku。SDS-PAGE分析结果显示,A5L蛋白主要以包涵体形式存在,最佳咪唑洗脱浓度为50 mmol/L。ELISA结果显示,制备的鼠抗A5L蛋白抗体最高效价达1∶205 440。【结论】本研究成功构建了重组质粒pET-28a-A5L,并在原核表达系统中成功表达重组蛋白A5L,制备的A5L多克隆抗体具有较好的免疫原性,研究结果为进一步探究MPXV A5L蛋白的生物学功能奠定基础。

猴痘病毒新型蛋白疫苗制备及其诱导抗体能力评估

目的构建有效的猴痘病毒(MPXV)和痘苗病毒(VACV)蛋白疫苗,并对其诱导抗体的活性进行初步评估。方法采用融合蛋白表达技术构建MPXV-A29-Fc和VACV-A27-Fc蛋白疫苗,酶联免疫吸附试验(ELISA)和蛋白质印迹法(Western blot,WB)体外检测蛋白结合抗体的能力,BALB/c雌性小鼠体内评估不同剂量蛋白疫苗诱导抗体的效价。结果MPXV-A29-Fc和VACV-A27-Fc蛋白均能与A29单克隆抗体结合,蛋白疫苗免疫小鼠后,不同剂量组中的小鼠血清均可交叉识别MPXV-A29和VACV-A27蛋白,三针免疫可诱导更强的交叉免疫应答,三针免疫后,不同的血清稀释度下,低剂量组的IgG抗体结合强度均与高剂量组相当,可达到高剂量组的免疫效果。结论本研究初步构建有效的MPXV和VACV新型蛋白疫苗MPXV-A29-Fc和VACV-A27-Fc,在小鼠体内可诱导特异性抗体反应和交叉抗体反应,可为猴痘病毒的疫苗设计提供新的思路和方法。

非典型猴痘一例

本文报道一例24岁非典型猴痘男性患者,3天前包皮及龟头出现环状丘疹,无发热、咽痛等系统症状,发病前有无保护男男性行为。皮肤科查体:包皮、龟头可见数个绿豆、黄豆大小的环状丘疹,中央凹陷可见结痂,躯干、四肢及头面无明显皮疹。猴痘病毒检测阳性。对症治疗后皮疹逐渐消退。本例猴痘患者仅出现生殖器皮损,与常规猴痘表现不相同,在临床中容易误诊。

猴痘病毒B20R蛋白原核表达及其多克隆抗体的制备

【目的】构建猴痘病毒B20R蛋白原核表达系统,并评价原核表达获得的融合蛋白免疫原性,为后续的猴痘病毒检测及新型治疗方法开发提供技术支撑。【方法】参照GenBank已公布的猴痘病毒基因组序列,按照大肠杆菌密码子偏好性对B20R基因DNA序列进行优化,人工合成B20R基因并将其分别克隆至表达载体pET-28a和pET-32a以构建原核表达载体;以构建的2种原核表达载体分别转化大肠杆菌BL21感受态细胞,经IPTG诱导表达后使用SDSPAGE和Western blotting检测融合蛋白B20R的表达情况。通过控制变量法优化融合蛋白B20R的诱导表达条件,使用Ni柱亲和层析进行纯化;以纯化的融合蛋白B20R经腹腔注射免疫昆明小鼠,定期采集昆明小鼠血样,采用ELISA检测血清抗体效价。【结果】将B20R基因克隆至表达载体pET-32a能成功构建原核表达载体pET-32a-B20R,经IPTG诱导后,可在大肠杆菌BL21感受态细胞中表达出融合蛋白B20R,且主要以不可溶的包涵体形式进行表达。融合蛋白B20R最佳诱导表达条件为37℃下以0.6 mmol/L IPTG诱导12 h,Ni柱亲和层析纯化的最佳咪唑洗脱浓度为40 mmol/L。以纯化的融合蛋白B20R接种免疫昆明小鼠,可在短时间内引起昆明小鼠产生强烈的体液免疫,且至免疫第42 d仍然维持较高的抗体水平,抗体效价高达1∶409600。【结论】构建的猴痘病毒B20R蛋白原核表达载体可在大肠杆菌BL21细胞中以包涵体形式成功表达出融合蛋白B20R,且融合蛋白B20R具有良好的免疫原性,可在短时间内引起昆明小鼠产生强烈的体液免疫。

广东地区人工驯养灵长动物的病毒组特征及猴痘病毒筛查

目的明确广东地区灵长动物携带病毒群落的结构特征,并评估重要人兽共患病毒猴痘病毒(MPXV)通过人工驯养灵长动物传入我国的风险。方法在广东省14个地级市的20个野生动物救护中心或动物园采集灵长动物样品,通过Illumina测序技术鉴定广东地区人工驯养灵长动物携带病毒组的结构特征;并通过荧光定量PCR方法,对MPXV进行检测,以确认MPXV经广东省人工驯养灵长动物传入的风险。结果共收集灵长动物口咽拭子和粪便489份。高通量测序结果显示,广东地区人工驯养灵长动物粪便中的病毒组结构复杂且具有地区差异,其中丰度最高的是Alphaflexiviridae和Virgaviridae的成员,其次为Parvoviridae和Genomoviridae成员。荧光定量PCR结果显示,广东省内野生动物救护中心或动物园的灵长动物全部为MPXV阴性。结论广东省人工驯养灵长动物携带病毒引起动物源性人兽共患病的风险极低,且MPXV通过广东省人工驯养灵长动物传入我国的风险几乎为零。

Rapid identification of full-length genome and tracing variations of monkeypox virus in clinical specimens based on mNGS and amplicon sequencing

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.

成都市锦江区污水中猴痘病毒的监测及基因特征分析

目的 调查成都市锦江区污水中猴痘病毒含量,并对其进行分型溯源。方法 2023年7~9月对成都市第九污水处理再生水厂未经消杀处理的污水进行24 h间断采样,共采集26个样本。用聚乙二醇沉淀法将水样从105 mL浓缩到0.6 mL后,提取核酸,用实时荧光聚合酶链反应(Polymerase Chain Reaction, PCR)法检测猴痘病毒,用猴痘病毒假病毒标准物质制备标准曲线计算相对浓度。对阳性的样本进行猴痘病毒全基因组扩增子测序。结果 8个样本检测出猴痘病毒阳性,检出率30.8%,均为西非分支Ⅱb C.1型。各分离株基因与报道的临床阳性株非常相似,与2022年猴痘疫情暴发的基因组密切相关。各分离株序列之间也非常相似,相似性为99%以上。结论 污水中的猴痘病毒都与报道的临床毒株高度相似,可以推定污水中的猴痘病毒均来自于患者或携带者的人体脱落物或排泄物。污水中猴痘病毒浓度与阳性报告病例的发生率基本呈正相关,表明污水监测是监测猴痘感染传播趋势的可行方法。

猴痘合并HIV感染一例

患者,男,21岁。10天前有男男高危性接触史。4天前出现发热,3天前肛周出现脓疱、溃疡,伴瘙痒。皮肤科查体:肛周见多发溃疡面。HIV感染2年。实验室检查:溃疡面组织液检查MPXV-qPCR(+)。诊断为猴痘。

Diagnostic approaches for monkeypox virus

Mpox(formerly Monkeypox)is a zoonotic infection caused by Monkeypox virus(MPXV).Since 2022,Mpox epidemics have occurred in many nonendemic countries and regions,leading the World Health Organization to declare a public health emergency of international concern.With the persistent transmission and evolution of MPXV,symptoms of Mpox have become milder,with some infections being asymptomatic.In addition,MPXV has become more contagious.Therefore,rapid and accurate diagnosis and screening of MPXV is vital to prevent and control MPXV epidemics.Here,we review and summarize the technical details,application scenarios,and the advantages and disadvantages of MPXV‐specific diagnostic methods.

猴痘抗病毒药物及疫苗研究现状

猴痘(Mpox)是一种由猴痘病毒(MPXV)感染引起的疾病,目前国际上没有针对猴痘病毒的特异性治疗药物,应用的抗病毒药物及疫苗既往主要用于治疗和预防天花等正痘病毒属相关疾病,抗病毒药物主要包括特考韦瑞、西多福韦、布林西多福韦等;疫苗主要使用ACAM2000、MVA-BN、LC16等,能够较好治疗和预防猴痘疾病的发生与发展。在无特效药物且无更有效、更安全疫苗的前提下,这些药物与疫苗可在早期治疗及预防中应用,以缓解猴痘病毒带来的临床症状,降低重症率和死亡率。本文从作用机制、临床试验、安全性等方面对抗病毒药物及疫苗进行论述,以期为我国猴痘的治疗和预防提供参考。

Monkeypox virus(MPXV):A Brief account of global spread,epidemiology,virology,clinical features,pathogenesis,and therapeutic interventions

The largest monkeypox virus(MPXV)outbreak of the 21st century occurred in 2022,which caused epidemics in many countries.According to WHO,physical contact with infected persons,contaminated surfaces,or affected animals might be a source of this virus transmission.A febrile sickness including few symptoms found in MPX disease.Skin rash,lesions,fever,headache,fatigue,and muscle aches symptoms were observed commonly for this disease.Animal and in vitro,studies have shown that the antiviral medications cidofovir and brincidofovir are effective against MPXV.The first-generation vaccinia virus vaccine was developed in 1960,and it helped to protect against MPXV with its side effects.A second-generation vaccination with limitations was launched in 2000.However,the CDC advised vaccinations for risk groups in endemic countries,including positive patients and hospital employees.The JYNNEOS vaccine,administered in 2 doses,also provides protection from MPX.This article presents concisely the most recent findings regarding epidemiology,genomic transmission,signs and symptoms,pathogenesis,diagnosis,and therapeutic interventions for MPXV,which may be helpful to researchers and practitioners.WHO declared that MPX was no longer a global health emergency due to its declining case rate,and a number of countries have reported new incidences.Further research-based investigations must be carried out based on the 2022 outbreak.

新形势下口岸防控输入性猴痘对策探讨

猴痘是由猴痘病毒(MPXV)引起的一种罕见的、散发的、天花样临床表现的急性传染病。2022年5月以来,全球报告了超过1000例猴痘实验室确诊病例,这是全球首次猴痘非流行国家和流行国家同时报告了多个猴痘病例和聚集性病例,对全球猴痘疫情的防控提出了新挑战。本文对猴痘的流行病学特征、临床表现、实验室诊断、本轮猴痘流行态势进行了综述,提出了旅行者个人防控措施建议和口岸应对本轮猴痘疫情的防控策略及建议。

猴痘病毒研究进展

猴痘是一种由猴痘病毒(MPXV)引起的症状和体征类似天花的人兽共患传染病。MPXV于1958年在实验室中的动物身上首次被发现,1970年在刚果民主共和国发现了第1例人感染MPXV,过去主要流行于中非和西非,但传染性和致病力均较弱。2022年以来,世界多国出现猴痘疫情,并呈现快速扩散态势。2022年7月23日,世界卫生组织(WHO)表示,猴痘疫情在70多个国家的蔓延是一种“非同寻常”的情况,将其列为全球紧急状态事件。论文围绕MPXV病原学(包括基因组特征、病毒复制、致病机理、分离培养和理化特性)、流行病学、致病性及实验室诊断方法等方面进行综述,以期为该病的科学预防和控制提供借鉴。

Monkeypox awareness,knowledge,and attitude among undergraduate preclinical and clinical students at a Malaysian dental school:An emerging outbreak during the COVID-19 era

Objective:To evaluate the awareness,knowledge,and attitude on monkeypox viral infection among preclinical and clinical dental students in Malaysia.Methods:A cross-sectional study was conducted among 229 preclinical and clinical dental students via an online self-reported questionnaire.The questionnaire included 3 items on awareness,15 items on knowledge,9 items on attitude and 2 open-ended questions.Data were presented as frequencies and percentages.Chi-square test was used to compare knowledge and awareness scores between preclinical and clinical dental students and content analysis was performed for open-ended responses.Results:Preclinical and clinical dental students were aware of the existence of monkeypox(89.5%and 94.4%,respectively),that the disease emerged in non-endemic countries(81.0%and 87.1%,respectively)and that it was declared a public health emergency of international concern by the World Health Organization(73.3%and 79.0%,respectively).Clinical dental students’overall knowledge level was significantly higher than preclinical dental students(P=0.014).Both preclinical(95.2%)and clinical(96.8%)dental students demonstrated positive attitudes toward monkeypox with no significant difference(P=0.736)noted between them.Three themes emerged from the open-ended questions:(1)reimplementation of nationwide lockdown,(2)impact on the economy and health,and(3)disruption to the educational system.Students also anticipated their face-to-face learning to be reduced should there be a new outbreak.Conclusions:Both preclinical and clinical dental students showed comparable awareness and attitudes,while the latter demonstrated greater satisfactory knowledge toward the re-emergence of monkeypox during the COVID-19 pandemic.Nonetheless,efforts to improve dental students’understanding of this alarming outbreak are required,to safeguard their health and minimise transmission.