目的 对两例无明确新型冠状病毒肺炎(coronavirus disease 2019,COVID-19,以下简称新冠肺炎)流行病学史,经多次核酸检测确诊的新冠肺炎病例实验室确诊过程进行总结分析,为新冠病毒核酸检测提供经验.方法 收集北京市2020年1-2月两例无明确新冠肺炎流行病史,且经多次核酸检测确诊的新冠肺炎病例的流行病学、临床表现等资料和实验室检测结果进行回顾性分析.结果 病例A,2020年1月19日发热伴全身疼痛、头痛,先后就诊2家医院,发热和肺炎病情未缓解.发病前有京外旅行史,2020年1月22日患者收入A2医院隔离治疗,诊断为"肺炎待查"并上报为新冠肺炎疑似病例.当日采集样本送北京市疾病预防控制中心(CDC)实验室进行荧光RT-PCR检测,N基因阳性.按当时《新型冠状病毒肺炎防控方案(第二版)》规定,实验室要求继续采样.1月23日再次采集样本,荧光RT-PCR检测ORF1ab和N基因阳性,该病例为新冠肺炎确诊病例.病例B,2020年2月3日发热,先后就诊3家医院,发热伴全身疼痛、头晕头痛.患者于2月9日在B3医院发热门诊就诊,并单间隔离治疗,上报为新冠肺炎疑似病例.2月10日和11日分别采集样本送北京市CDC实验室进行荧光PCR检测,ORF1ab和N基因阴性,但2月10日 样本的N基因在反应后期显示出荧光值小于判定阈值线的微弱信号.2月12日再次采集样本检测,ORF1ab和N基因阳性,该病例为新冠肺炎确诊病例.结论 因新冠肺炎临床表现无特异性,实验室病原学的核酸检测非常重要;如果新冠肺炎疑似病例首次样本检测结果阴性或可疑,建议再次采集多种类样本进行检测,以便尽可能早诊断.

目的 探讨DNA条形码技术鉴定本地郊区常见鼠形动物的可行性.方法 收集134份本地常见鼠形动物样本,PCR扩增线粒体细胞色素C氧化酶亚基Ⅰ基因,测序后进行序列比对,并计算物种种内和种间遗传距离,采用邻接法构建系统发育树.结果 序列比对结果显示共有黑线姬鼠22只、中华姬鼠19只、小家鼠25只、褐家鼠39只、社鼠19只、白腹鼠未定种4只、山西(鼲)2只、大仓鼠2只、川西缺齿鼩鼱2只.其中,有2份现场形态学鉴定为小家鼠的样本,经分子方法鉴定为褐家鼠,提示现场鉴定有误.系统发育分析显示,各物种种内遗传距离远小于种间遗传距离;所采集样本在进化树上形成8个高度支持的独立分支,与形态学鉴定结果基本一致.结论 DNA条形码技术能有效鉴定本地鼠形动物种类,有助于提高本地鼠形动物鉴定的准确性.

Objective:To study the clinical features and epidemiological characteristics of the first local case of severe fever with thrombocytopenia syndrome (SFTS) on October 2021 in Beijing, so as to provide basis for formulating the prevention and control strategy.Methods:The epidemiological characters of the case was analyzed by epidemiological investigation methods. Real time fluorescent quantitative PCR and ELISA were used to detect nucleic acid of severe fever with thrombocytopenia syndrome virus (SFTSV) and antibodies against the virus. The vectors and host animals were also investigated.Results:The investigation results indicated that the patient haven’t leave from Beijing for many years. Nucleic acid of SFTSV and IgM antibody were detected in the serum sample and the viral load was 1.9×10 4 TCID 50/ml. The 5 ticks captured during vector survey were negative for SFTSV nucleic acid. The blood sample of the pet dog was also negative for both SFTSV nucleic acid and antibody against the virus. Conclusions:The SFTS case on October 2021 was the first local case reported in Beijing. Surveillance for SFTS and vectors should be strengthened.

目的 调查北京市鼠形动物巴尔通体感染状况和遗传特征.方法 采用夹夜法在北京市16个区捕获鼠形动物,使用PCR方法检测巴尔通体gltA基因,测序后进行系统发育树分析.结果 共采集7属9种鼠形动物样本448份,检测到巴尔通体阳性样本46份,总阳性率为10.3％,且阳性率在鼠种间、地区间和生境间的差异均具有统计学意义(x2鼠种=114.980,x2地区=22.133,x2生境=96.466,P均＜0.001).系统发育树分析显示,40份样本序列位于进化树上的8个不同分支,提示北京市鼠形动物巴尔通体感染存在遗传多样性.结论 巴尔通体在北京市鼠形动物中广泛存在,并具有遗传多样性,本地人群存在一定的感染风险.

目的 分析北京市2009-2019年恙虫病的流行特征,掌握疫情变化趋势,为恙虫病的防控提供科学依据.方法收集北京市2009-2019年报告的恙虫病病例资料,使用WPS office表格工具进行数据处理及图表绘制,SPSS statistics 21.0进行统计分析,SatScan 9.5进行时空扫描分析,Mapinfo 9.5进行空间分布时空扫描结果可视化.结果 2009-2019年北京市共报告恙虫病病例1 060例,年平均发病率为0.49/10万.2009-2015年发病率呈上升趋势,2015-2018年呈下降趋势,2018-2019年病例报告数再次上升.北京市恙虫病病例报告涉及全北京10个区(县),其中发病率最高的地区为平谷区,平谷区病例报告数占发病总数的90.19％(956/1 060).恙虫病发病高峰期在秋冬季,9-11月报告病例数占发病总数的98.02％(1 039/1 060),10月为发病高峰.50～59岁是主要的发病年龄,占发病总数的32.83％(348/1 060),不同年龄段对疾病发生率的差异有统计学意义(X2=8480.0,P＜0.001),发病人群中男女各半,各年龄段发病人群中因性别产生的差异无统计学意义(X2=0.795,P=0.373).农民为恙虫病发病率最高人群,占发病总数的75.00％(795/1 060).结论 北京市恙虫病发病水平较高,发病季节和地区较为集中,发病人群相对固定,应在恙虫病高发季节对重点人群进行监测,采取健康教育等预防干预措施,降低恙虫病的发病水平.

目的 对北京某医院就诊的一例人感染猪链球菌患者的血培养分离菌株,并进行分子分型与耐药检测分析.方法 将患者血培养阳性物接种于哥伦比亚血琼脂平板,分离培养后提取菌株DNA,采用多位点序列分型(MLST)确定其分子型别;采用微量肉汤稀释法进行12种抗生素的药敏试验.结果 该菌株的多位点序列分型为ST1型.药敏试验结果表明:该菌对青霉素、左氧沙星、美罗培南、万古霉素、克林霉素、头孢噻肟、利奈唑胺、红霉素和头孢吡肟9种常用抗生素敏感;对氧氟沙星中度敏感;对四环素和氯霉素耐药.结论 该猪链球菌为ST1型,是引起脑膜炎的主要型别;对9种常用抗生素菌敏感,可做为临床治疗的参考用药.

Objective:To manage and analyze an suspected imported Chikungunya fever case in 2017 in Beijing.Methods:Epidemiological investigation was performed on the suspected case. The saliva and urine samples were collected and nucleic acid of Chikungunya fever virus was detected by fluorescent real-time PCR.Results:The results of nucleic acid tests indicated that the nucleic acid of Chikungunya virus was positive, confirming the case to be an imported Chikungunya case. The hotel was instructed for disinfection measures and the knowledge on disease prevention and control was publicized.Conclusions:There was potential risk of Chikungunya fever outbreak in Beijing. The surveillance should be strengthened.

2019年11月,北京市确诊2例输入性肺鼠疫病例,这是建国后肺鼠疫首次输入北京市,也是肺鼠疫首次进入千万人口的大城市.该起疫情具有远距离传播、密切接触者众多且分散等特点,本文简单介绍了该起疫情的处置过程并重点探讨了城市鼠疫防控中的问题.

目的 评价3种浓度核酸酶消化在蜱虫宏基因组学研究中的作用.方法 经过3种浓度的核酸酶处理蜱虫样本后,采用Illumina Miseq二代测序仪测序,分析比较蜱虫宏基因组学物种的种类及优势物种reads的丰度,初步评价不同终浓度(0 U/μl、0.05 U/μl和0.2 U/μl)的核酸酶处理样本对宏基因组学结果的影响.结果 采用核酸酶处理虽然降低了可检出物种的种类,但能够有效提高大部分优势物种的检出比例,尤其是立克次体属、不动杆菌属和Andromedavirus病毒属等物种的检出率;低剂量的核酸酶处理组的提高效果比高剂量处理组和无核酸酶处理组都要好.结论 采用低剂量核酸酶(0.05 U/μl)处理蜱虫样本会提高宏基因组研究中发现立克次体及病毒等病原的几率,为蜱虫及其他生物样本的深度测序过程中样本富集步骤的优化提供了一个初步的实验依据.

Background: Malaria is a parasitic disease that threats human health seriously. Since 2017, there have been no reports of local malaria infections in the mainland of China, and all malaria cases were imported. An early and accurate diagnosis is critical for the prognosis of malaria cases and for the control of local epidemics. Microscopic examinations, antigen and nucleic acid detection are the most common diagnostic methods, however, they still have not met the desire for diagnosis of malaria. As a new method, next generation sequencing (NGS) has become an indispensable technology in infectious diseases diagnosis, however, the applications of this technique to the diagnosis of parasites such as malaria parasites were rarely reported. Methods & Materials: The blood samples of an imported patient from Africa who was suspected to be infected with malaria were collected and processed for NGS detection using Illumina Miseq Next-Generation Sequencer. The reads were filtered by quality and host contaminating reads were filtered by mapping to the Human reference genome (hg19) with CLC genomic workbench (QIAGEN). The malaria parasites were classified at the species levels using lowest common ancestor analysis (LCA) with MEGAN6. At the same time, we performed microscopic examination and real-time fluorescent PCR typing of Plasmodium. Results: NGS successfully identified the patient as malaria parasites infection. The results of NGS showed that the dominant species in the blood was Plasmodium falciparum. Further analysis revealed that it has the highest homology with Plasmodium falciparum 3D7. Results from microscopic examination also showed that the case was infected with Plasmodium falciparum, at the same time, there was a very small number of Plasmodium malaria. The results of real-time fluorescent PCR showed that the nucleic acid of P. falciparum was positive, and the nucleic acids of P. vivax, P. malaria and P. ovaria were negative. Conclusion: NGS can successfully detect malaria parasites infection and the results were consistent with real-time fluorescent PCR and microscopic examination. Although microscopy seemed a bit more sensitive, considering the detection of major pathogens and especially pathogen traceability with less reliance on clinical symptom information, NGS is more promising for pathogen screening in imported fever cases.

2019年11月,北京市经实验室检验确诊2例鼠疫病例,根据患者的流行病学史与临床表现确认为输入性肺鼠疫,这是建国后肺鼠疫首次输入北京市.该起疫情中实验室检测发挥了重要作用,本文简单介绍此次疫情中的实验室检测过程并重点探讨鼠疫实验室检验中检测方法与检测试剂的相关问题.

目的 探讨MLVA分型对于鉴别布鲁氏菌病复发与重复感染的意义.方法 对初次发病与二次发病的布病患者进行血培养,分离菌株,对分离菌株做布鲁氏菌种属鉴定与MLVA-16分型鉴定,分析两次发病分离到的菌株的分型结果及各位点的差异.结果 收集到4例二次发病的布病患者的血培养物,分离得到8株布鲁氏菌,经MLVA-16分型鉴定为7个基因型,1例两次发病时间间隔较短的患者的2株分离株的MLVA-16分型一致,没有差别,提示为复发.而另外3例两次发病时间间隔较长的患者的6株分离菌株,其MLVA-16分型的基因型不同,分别各有一个位点的差异,提示为重复感染.结论 MLVA分型对于布鲁氏菌病的复发与重复感染具有一定的鉴别意义.

Objective To investigate the community composition,species diversity and density of rodents and to explore the infection of Yersinia pestis in Beijing during 2011 to 2018. Methods The surveillance was conducted in the districts of Mentougou,Yanqing,Huairou,Miyun and Shunyi. The species were identified and the density was calculated using the trap -at-night method . The blood samples were collected from the heart of the rodents captured alive by cage-trap method and the anti-Yersinia pestis F1 antibodies were tested using serological method referred to Diagnostic Criteria for plague (WS- 279) . Results There were 1 454 rodents captured during 2011 to 2018. These rodents were classified into eight species. Among them,Niviventer confucianus (43. 5％) ,Apodemus peninsulae (33. 6％) and Apodemus agrarius (11. 6％) could be infected with and spread Yersina pestis. The density of rodents by year changed from 2. 52％ to 6. 02％ (P＞0. 05) ,and the density by district changed from 4. 01％ to 6. 22％ (P＞0. 05) . The highest density (17. 25％) was found in October. The serological F1 antibodies were all negative in 827 blood samples. Conclusion The three rodents with highest density could be infected by Yersinia pestis which are Niviventer confucianus,Apodemus peninsulae and Apodemus agrarius. But no infection of Yersinia pestis are founded by serological test method .

目的 对北京地区分离自布病患者血液的2株布鲁菌进行多位点序列分型(Multilocus sequence typing,MLST)及其遗传学特征进行研究,为布病的预防和控制提供科学依据.方法 应用布鲁菌属特异性PCR(BCSP31-PCR)和种/型特异性PCR(AMOS-PCR)对分离的2株布鲁菌进行鉴定,采用MLST技术对2株布鲁菌分离株的19个管家基因、1个外膜蛋白基因及1个基因间区的序列进行测定,与MLST数据库中的等位基因序列进行比对,确定菌株的等位基因谱及9位点和21位点序列型(sequence type,ST).采用聚类分析法研究2株菌ST型与各种布鲁菌已知ST型的遗传进化关系.结果 2株分离株经BCSP31-PCR鉴定为布鲁菌属细菌,AMOS-PCR鉴定为非羊种、非牛种1、2、4型、非猪种1型、非绵羊附睾种布鲁菌;MLST分析显示,2株菌的等位基因编号(gap /aroA/glk /dnaK /gyrB /trpE/cobQ/int-hyp /omp 25/prpE/caiA/csdB/soxA/leuA/mviM/umC/fbaA/ddlA/putA/mutL /acnA)分别为2、1、2、2、1、3、1、4、1、13、2、2、2、2、1、1、3、7、6、2、3和2、1、2、2、1、3、1、4、29、13、2、2、2、2、1、1、3、7、6、2、3,比对MLST数据库,结果显示这2株细菌的等位基因编号组合未见报道,为新ST型.聚类分析显示这2种新的ST型与牛种布鲁菌ST型处于同一分支.结论 北京地区分离的2株布鲁菌为新ST型牛种布鲁菌,已被MLST数据库确认,分别命名为ST74和ST75(9位点序列型)与ST108和ST109(21位点序列型).与同属牛种布鲁菌的ST型遗传关系最近,该结果为北京地区布病的预防和控制提供了科学依据.

目的 将黄热病毒实验诊断过程中的生物安全和生物安保风险降到最低.方法 识别可疑黄热病病例样本在检测前、检测中及检测后的生物危害风险,依据风险分析和评估结果制定并采取风险防控措施.结果 在检测前进行风险识别和分析,优化病原检测所需场所、仪器、材料及人员,确定操作规程及人员防护措施;对检测中样本开启、处理、分装、暂存及检测后废弃物处理、实验环境消毒、样本保藏及上送等各阶段开展风险识别及风险再评估,持续优化、完善防控措施.结论 本研究为黄热病及其他蚊媒传播传染病的风险评估和应对措施制定提供了参考和依据.

Objective To explore the seasonal distribution characteristics of brucellosis in Beijing from 2004 to 2015 so as to provide evidence for brucellosis prevention and control.Methods Concentration degree and circular distribution methods were used to conduct temporal cluster analysis of the incidence of brucellosis in Beijing during 2004-2015.Results The M value was 0.30,showing the seasonal concentration distribution characteristics.Circular distribution analysis showed that the peak incidence of brucellosis was on May 17th and the epidemic period was from February 16 to August 15.Conclusions The incidence of brucellosis has certain seasonal distribution features,showing the high incidence in spring and summer and the low incidence in fall and winter.

We report the first imported case of Rift Valley fever (RVF) in China. The patient returned from Angola, a non-epidemic country, with an infection of a new reassortant from different lineages of Rift Valley fever viruses (RVFVs). The patient developed multiorgan dysfunction and gradually recovered with continuous renal replacement therapy and a short regimen of methylprednisolone treatment. The disordered cytokines and chemokines in the plasma of the patient revealed hypercytokinemia, but the levels of protective cytokines were low upon admission and fluctuated as the disease improved. Whole-genome sequencing and phylogenetic analysis revealed that the imported strain was a reassortant comprising the L and M genes from lineage E and the S gene from lineage A. This case highlights that RVFV had undergone genetic reassortment, which could potentially alter its biological properties, cause large outbreaks and pose a serious threat to global public health as well as the livestock breeding industry.

To identify and analyze the virulence of a bacteria strain isolated from the blood of a patient with suspected Streptococcus suis (S.suis) infection in a hospital of Beijing,we inoculated the bacteria strain isolated from the blood of the patient to the Columbia with sheep blood agar plate,after Gram staining and microscopical examination,serum agglutination test,VITEK 2 Compact microbial identification system test and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) test,S,suis species specific gene 16SrRNA,S.suis species serotype 2 specific virulence gene capsule polysaccharide 2J (cps2J) and virulence gene muramidase-released protein (mrp),hemolysin (sly),extracellular factor protein (ef),glutamate dehydrogenase (gdh) genes,fibronectin-binding protein (fbps),glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes and virulence correlated gene orf2 were further detected by PCR.Results showed that the suspicious bacteria strain of S.suis was identified as S.suis type 2 (S.suis 2) by conventional methods,MALDI-TOF-MS and PCR.PCR results showed that cps2J,sly,ef,gdh,fbps,gapdh and or f2 genes were positive,and mrp gene was negative.In conclusion,the bacteria strain isolated from the patient's blood is sly+/ef+/mrp-virulent S.suis 2.

Descriptive epidemiological method, circular distribution methods and SaT Scan 9.4 software were used to analyze the temporal and spatial distribution,and other epidemiological characteristics of dengue fever in Beijing from 2006 to 2016. Arcgis software was used to analyze the spatial distribution of the cases. There were 131 cases of dengue fever enrolling in the present study in Beijing. Circular distribution analysis showed that the peak of dengue fever was on August 25 and the epidemic period of dengue fever was from April 28 to December 23. Dengue cases clustered as a circular region, with radius 10.20 km, centered geographic coordinates 39.98 N, 116.41 E. These dengue fever cases were mainly imported from Southeast Asia and South Asia. The onset date of illness, infection site and address showed temporal and spatial aggregation. The main occupations included staffs, students, housekeeping personnel, unemployed and business servants. The results obtained in the present paper provide scientific basis for the prevention and control of dengue fever.

Objective To describe the epidemiological characteristics,the management and control measures of three confirmed imported patients of Zika virus disease in Beijing.Methods The patients were interviewed,and the information on epidemiological history and clinical manifestation was acquired.The RT-PCR method was employed to test the blood,urine and saliva samples of the patients.The control measure included the quarantine of the patients,disinfection,and health education.Results Morbilliform rash was a common main clinical symptom of the three patients,accompanied by low-grade fevers and conjunctival congestion.All three patients got infected and suffered an onset in Caracas,the capital of Venezuela.They arrived in Beijing in 1,3,7 days after onset and were quarantined from mosquito and hospitalized after self-declare to a quarantine officer.The urine samples of the three patients were positive for Zika virus detected by real-time PCR.Conclusions This report summarized related information of the three imported cases of Zika virus disease in Beijing and discussed the related issues in control measures.And it will be helpful to the management of imported Zika virus disease and other mosquito-borne diseases in the future.