耶尔森菌,鼠疫

摘要： 目的了解内蒙古自治区(简称内蒙古)15株鼠疫耶尔森菌(简称鼠疫菌)差异区段(DFR)分型情况。方法对2019年内蒙古高原长爪沙鼠鼠疫自然疫源地鄂托克旗、达茂旗和松辽平原达乌尔黄鼠鼠疫自然疫源地正镶白旗分离的15株鼠疫菌,采用PCR技术扩增,并进行DFR分型。结果来自鄂托克旗的2株鼠疫菌共同缺失DFR位点1、6、7、13、15、16、17、18、23,来自达茂旗和正镶白旗的13株鼠疫菌共同缺失DFR位点1、6、7、12、13、15、16、17、18、23。通过与DFR位点分布和基因型结果比对,分别归类为基因型G17和G20。结论内蒙古高原长爪沙鼠鼠疫自然疫源地和松辽平原达乌尔黄鼠鼠疫自然疫源地鼠疫菌DFR分型与历史菌株基因型相同。

摘要： 目的 了解内蒙古自治区(简称内蒙古)15株鼠疫耶尔森菌(简称鼠疫菌)差异区段(DFR)分型情况.方法 对2019年内蒙古高原长爪沙鼠鼠疫自然疫源地鄂托克旗、达茂旗和松辽平原达乌尔黄鼠鼠疫自然疫源地正镶白旗分离的15株鼠疫菌,采用PCR技术扩增,并进行DFR分型.结果 来自鄂托克旗的2株鼠疫菌共同缺失DFR位点1、6、7、13、15、16、17、18、23,来自达茂旗和正镶白旗的13株鼠疫菌共同缺失DFR位点1、6、7、12、13、15、16、17、18、23.通过与DFR位点分布和基因型结果比对,分别归类为基因型G17和G20.结论 内蒙古高原长爪沙鼠鼠疫自然疫源地和松辽平原达乌尔黄鼠鼠疫自然疫源地鼠疫菌DFR分型与历史菌株基因型相同.

摘要： Objective To identify the causes of nonspecific bands in the detection of a industry standard caf1 gene by polymerase chain reaction(PCR),and to propose a solution to this problem. Methods A total of 112 strains were selected for the experiment, including 40 strains of Yersinia pestis, 72 strains of non-Yersinia pestis;DNA was extracted,and caf1 gene was amplified by PCR;seven non-specific strips were recovered,purified and TA cloning and sequencing; the primer of the caf1 gene was redesigned and validated using all of the strains. Results Using the industry standard caf1 gene primer,DNAs of 40 Yersinia pestis and 72 non-Yersinia pestis were amplified by PCR, 58 non-Yersinia pestis could be amplified with non-specific bands, they were about 400, 500, 600, 700, 800, 900, 1 000 bp. By TA cloning and sequencing, the non-specific bands in the downstream of the industry standard caf1 primer and its reverse complement were amplified. Using the new designed caf1 primer to amplify, 72 non-Yersinia pestis strains showed no non-specific bands. Conclusion Non-specific bands has been amplified in the screening of Yersinia pestis using the primer of the industry standard caf1, and the new caf1 primer can effectively avoid this problem and improve the accuracy of detection.%目的 查明行业标准推荐caf1基因引物(简称行标caf1引物)进行PCR检测时,在一些非鼠疫菌上出现非特异性条带的原因,并提出解决办法.方法 共选取112株菌进行试验,其中鼠疫菌40株、非鼠疫菌72株;提取菌株DNA,并对caf1基因进行PCR扩增;选择出现非特异性条带的7株菌株,进行切胶回收、纯化以及TA克隆测序;针对caf1基因重新设计引物,对被试菌株进行验证.结果 使用行标caf1引物扩增40株鼠疫菌,均未出现非特异性条带,扩增72株非鼠疫菌,有58株扩增出400、500、600、700、800、900、1000 bp的非特异性条带.经克隆和测序,在非特异性产物的基因序列中均出现行标caf1引物的反向引物序列及反向引物的反向互补序列.重新设计caf1引物,扩增72株非鼠疫菌,均未出现非特异性条带.结论 行标caf1引物在筛查鼠疫菌时会出现非特异性条带,使用新caf1引物可有效避免这一情况,提高检测的准确性.

摘要： Objective To study the biological characteristics and epidemiological significance of Yersinia pestis in Chengduo County of Qinghai Province,in order to provide scientific basis for plague prevention and control in this area.Methods Thirty one strains of Yersinia pestis isolated from Chengduo County of Qinghai Province from 1980 to 2011 were selected as study subjects.Biochemical test,virulence factors evaluation [Fra1 (F1),pesticin Ⅱ (Pst Ⅱ),virulence antigen (VW),pigmentation (Pgm)] and different region (DFR) genotyping were carried out.Nineteen of the 31 strains Yersinia pestis were selected according to different time,different areas and different hosts to determine their toxicity in mice,MLD ≤ 10 000 was strong toxic strain,10 000 ＜ MLD ＜ 100 000 was moderate toxic strain.Results Among thirty one strains of Yersinia pestis,23 strains were isolated from human,the Himalaya marmot and its fleas and lice,and their biological type was classical,biochemical type was Qinghai-Tibet plateau;21 strains genotype was type 5,1 was type 16,1 was type 32,and they contained all four kinds of virulence factors (F1,Pgm,Pst Ⅱ,VW),and toxicity test showed all strains (14) were strong toxic strains.The rest 8 strains of Yersinia pestis isolated from the Microtus fuscus and its fleas,and their biological type was Microtus,biochemical type was Chuanqing plateau;they could produce F1 and Pgm,of which 87.5％ (7/8) strains could produce Pst Ⅱ,but could not produce VW antigen factor,the genotype was 14,and the toxicity results showed that they were strong (3)and moderate (2) toxic strains.Conclusion The strains separated in Chengduo County of Qinghai Province from 1980 to 2011 have the pathogen characteristics of Qinghai-Tibet plateau plague,they are mainly strong toxic strains;the work on prevention and control of plague should not be neglected.%目的 探讨青海省称多县鼠疫菌株生物学特点及流行病学意义,为该地区的鼠疫防治提供科学依据.方法 以1980-2011年称多县分离的31株鼠疫菌株作为实验对象,进行生化试验、毒力因子鉴定[荚膜抗原(F1)、色素沉着因子(Pgm)、鼠疫杆菌素Ⅰ(PstⅠ)、毒力抗原因子(VW)]、差异片段(DFR)分型,并根据不同时间、不同地区、不同宿主选取19株菌株进行小白鼠毒力测定,以最小致死量(MLD)表示,MLD≤10 000为强毒菌,10 000＜ MLD＜100 000为中等毒力菌.结果 31株被试菌株中,有23株是从喜马拉雅旱獭及其媒介蚤、虱和人体分离的,生物型为古典型,生化型为青藏高原型;21株基因型为5型,1株为16型,1株为32型;F1、Pgm、PstⅡ、VW 4种毒力因子俱全,毒力测定结果为强毒菌(14株).有8株是从青海田鼠及其媒介蚤分离的,生物型为田鼠型,生化型为川青高原型;均能产生F1及Pgm,其中87.5％(7/8)的菌株可以产生Pst Ⅱ,均不能产生VW抗原因子;基因型为14型,毒力测定结果为强毒菌(3株)、中等毒力菌(2株).结论 青海省称多县1980-2011年鼠疫菌株具备青藏高原鼠疫病原体特性,鼠疫菌毒力强,鼠疫防治工作不容松懈.

摘要： 目的 探讨青海省祁连县鼠疫菌株生物学特点及流行病学意义,为该地区的鼠疫防治提供科学依据.方法 对1958-2011年青海省祁连县分离的67株鼠疫菌进行生化试验、毒力测定、毒力因子鉴定、质粒分析、鼠疫菌差异区段(different region,DFR)分型等研究.结果 根据生化指标,67株被试菌株中48株为青藏高原型,15株为祁连山型,其余4株与青海省疫源地生态型菌株均不相同;DFR分为8个基因组型,分别为8型(42株)、44型(15株)、5型(4株)、7型(2株)、19型(1株)、30型(1株)、32型(1株)、34型(1株);95.52％ (64/67)的被试菌株携带相对分子质量为6×106、45×106、52×106的质粒;85.07％(57/67)的鼠疫菌4个毒力因子俱全;96.00％(48/50)的鼠疫菌为强毒菌.结论 青海省祁连县分离的鼠疫菌具备青藏高原鼠疫病原体特性,鼠疫菌的毒力强,因此,要加大鼠疫监测力度和鼠疫防治知识宣传力度,严防动物间鼠疫波及人间.%Objective To investigate the biological characteristics and epidemiological significance of Yersinia pestis strains in Qilian County,Qinghai Province,in order to provide a scientific basis for plague prevention and control.Method Totally 67 strains were separated from kinds of host in Qilian County,Qinghai Province from 1958 to 2011,to do biochemical test,toxicity test,virulence factors evaluation,plasmid analysis and different region (DFR) genotyping.Results According to biochemical typing,48 of the 50 strains tested were Qing-Tibet Plateau ecotype,15 were Qilian Mountain ecotype,and the remaining 4 were different ecotypes from the plague foci in Qinghai plateau.The strains had 8 genomovars,and were given priority to genomovar8 (42 strains),secondly,genomovar44 (15 strains),genomovar5 (4 strains),genomovar7 (2 strains),genomovar19 (1 strain),genomovar30 (1 strain),genomovar32 (1 strain),and genomovar34 (1 strain).A proportion of 95.52％ (64/67) of the strains contained 3 kinds of plasmid-6 × 106,45 × 106,and 52 × 106;85.07％ (57/67) contained all the four virulence factors,and 96.00％ (48/50) were velogenic strains.Conclusion The strains separated in Qilian County,Qinghai Province have the characteristics of Qinghai-Tibet Plateau plague's pathogen and have strong toxicity,so we should enhance the plague monitoring and give more publicity to plague prevention to prevent animal plague spreading to human.

摘要： 在鼠疫自然疫源地中,鼠疫菌与其宿主动物和媒介生物在一定地理景观内以及特定区域的生物群落中构成了相互依赖又相互制约的生态系统.目前,鼠疫菌的保存机制主要概况为:①宿主动物、蚤(蜱等)保存学说;②鼠疫菌自身突变保存学说;③蚤-宿主-蚤保存学说;④土壤保存学说;⑤鸟类及非生物因素保存学说;⑥其他耶尔森菌突变保存学说.文中针对鼠疫菌在疫源地保存机制的研究进展做以下综述.%Yersinia pestis,host animals and vector organisms in a particular area within a certain geographical landscape,and biological communities are in the form of interdependence and mutual restriction of ecological system in the plague natural foci of plague.Currently,the preservation of Yersinia pestis mechanism mainly including:①Animal and fleas (tick) preservation theory;②Yersinia pestis own mutation preservation theory;③Fleas-host-fleas preservation theory;④Theory of soil preservation;⑤the birds and abiotic factors of preservation theory;⑥Theory of other Yersinia preservation theory and so on.The author summarized the research progress of the preservation mechanism of Yersinia pestis in the focus area.

摘要： Objective To type Yersinia pestis isolated from Gansu Province,and to study the trend of diffreent strains in different administrative regions and different years.Methods Totally 193 strains were enrolled in this study,including 9 strains of Ganning Dauricus type,18 strains of Aerjin type,45 strains of Qilian type and 121 strains of Qingzang type.These strains were genotyped by clustered regularly interspaced short palindromic repeats (CRISPR),and genotypes were named according to international standard.Genotyping by CRISPR in different administrative regions and different years of Gansu Province was explored.Results Two clusters (Ca7 and Cb4),including four genotypes (genotypes 7,22,24 and 26) were classified by CRISPR.From the point of view of origin,genotype 24 was the main genotype in Akesai 36.36％ (16/44),Subei 36.17％ (17/47),Yumen 50.00％ (5/10) and Su'nan 38.67％ (29/75);the main genotype of Xiahe and Huining was genotype 26 (4/7);the main genotype of Shandan was genotype 22 (1/1).From the point of view of time,the main genotype of Yersiniapestis in Gansu Province during the years of 1960-1969,1970-1979 and 1980-1989 was genotype 26 [53.33％ (8/15),60.00％ (6/10) and 48.28％ (14/29)];the main genotype was genotype 22 [40.91％ (18/44)] during the years of 1990-1999;and the main genotype was genotype 24 [43.16％ (41/95)] during the years of 2000-2009.Conclusion Four genotypes of Yersiniapestis in Gansu Province are quite different in different administrative regions and different years.%目的 对甘肃省鼠疫菌株进行基因分型,分析不同生态型鼠疫菌株在不同行政地区、不同时间的流行趋势.方法 运用规律成簇的间隔短回文重复序列(CRISPR)对甘肃省甘宁黄鼠型(9株)、阿尔金型(18株)、祁连型(45株)及青藏型鼠疫菌株(121株)进行分型及遗传进化关系分析,分型按国际通用标准命名.并探讨甘肃省不同行政地区和不同流行时间CRISPR基因分型的变化情况.结果 甘肃省鼠疫菌株被分为2个CRISPR基因簇(Ca7和Cb4),4个基因型(7、22、24及26型).从来源地看,阿克塞、肃北、玉门、肃南均以24型[36.36％ (16/44)、36.17％(17/47)、50.00％(5/10)、38.67％(29/75)]为主基因型,夏河、会宁均以26型(4/7)为主基因型,山丹以22型(1/1)为主基因型.从时间上看,甘肃省1960-1969、1970-1979、1980-1989年鼠疫菌均以26型[53.33％(8/15)、60.00％(6/10)、48.28％(14/29)]为主基因型,1990-1999年以22型[40.91％(18/44)]为主基因型,2000-2009年以24型[43.16％(41/95)]为主基因型.结论 甘肃省鼠疫菌基因型在不同行政地区、不同时间流行趋势不同.

摘要： Objective To investigate the CRISPR genotypes (clusters) and regional distribution of Yersinia pestis in Qinghai-plateau. Methods One hundred and two isolates of Y. pestis isolated from human plague patients, host animal and insect vectors from Qinghai-plateau were selected. The DNAs were extracted using the traditional sodium dodecyl sulfate decomposition and phenol-chloroform method. Three CRISPR loci YPa, YPb and YPc of 102 isolates of Y. pesits were amplified and sequenced, and then the CRISPR sequence analysis was carried out by comparing the latest published CRISPR spacer dictionary and the NCBI database to identify the spacer and spacer array. CRISPR genotyping of isolates of Y. pesits were finally conducted according to the polymorphism of the spacer arrays and the regional distribution pattern of isolates of Y. pesits in Qinghai-plateau was described. Results Forty spacers including 22 of YPa, 13 of YPb and 5 of YPc were observed among 102 isolates of Y. pestis in Qinghai-plateau, of which 5 spacers (a1', a103, a104, b4' and b4' ') were firstly identified. Meanwhile, 16, 10, and 5 different spacer arrays were obtained in YPa, YPb and YPc respectively, including 11 new spacer arrays detected in this study. One hundred and two isolates were divided into 24 CRISPR genotypes and classified into 9 CRISPR clusters (Cb4, Cb4', Cb2, Ca37, Ca7, Ca7', CaΔ5', Ca35' and Cc3'). Each dominant cluster presented significant aggregation geographically: Ca7 were found in Yushu, Nangqian, Chenduo, Zaduo, Zhiduo and Qumalai countries. Ca7' were found in Xunhua, Tongren, Zeku, Tongde, Maqin and Guinan countries. CaΔ5' were restricted to Qilian, Gangcha, Menyuan and Datong countries. CaΔ35' were found in Huangyuan, Haiyan, Gangcha, Tianjun, Delingha, Wulan, Doulan, Gonghe, Xinghai, Guide and Tongde countries. Conclusion CRISPR-based genotyping analyses showed complicated population of Y. pestis in Qinghai-plateau. Four clusters, Ca7, Ca7', CaΔ5' and Ca35' were the most epidemic dominant four clusters and presented obvious regional distribution patterns, which instructed us to strengthen the surveillance and prevention and control by CRISPR-genotyping technique.%目的 研究青海高原鼠疫耶尔森菌(鼠疫菌)的CRISPR基因型(类群)及其地区分布.方法 选取1954—2011年间青海高原境内各县/市从人鼠疫患者、宿主动物及媒介昆虫体内分离的鼠疫菌102株,采用传统的十二烷基磺酸钠裂解和苯酚-氯仿法提取鼠疫菌DNA.分别对YPa、YPb和YPc等3个CRISPR位点进行PCR扩增、测序,然后将所测得CRISPR序列与文献最新报道的CRISPR Dictionary和NCBI数据库检索比对,以鉴定CRISPR spacer阵列.最后根据CRISPR spacer阵列的多态性对青海高原鼠疫菌进行基因分型,并描绘其地区分布特征.结果 102株鼠疫菌共发现40种spacer,包括YPa 22种、YPb 13种、YPc 5种,其中有5种新spacer,分别为a1'、a103、a104、b4'和b4';Ypa、Ypb和Ypc分别鉴定出16、10和5种不同的spacer阵列,新发现的spacer阵列为11种.102株鼠疫菌可被分为24个基因型,共归为9大CRISPR类群,分别为Cb4、Cb4'、Cb2、Ca37、Ca7、Ca7'、CaΔ5'、Ca35'和Cc3',其中Ca7、Ca7'、CaΔ5'、Ca35'呈现出显著的聚集性特征:Ca7主要分布在玉树、囊谦、称多、杂多、治多和曲麻莱;Ca7'分布于循化、同仁、泽库、同德、玛沁和贵南;CaΔ5'集中在祁连、刚察、门源、大通;Ca35'分布在湟源、海晏、刚察、天峻、德令哈、乌兰、都兰、兴海、共和和贵德.结论 青海高原鼠疫菌CRISPR种群结构复杂,Ca7、Ca7'、CaΔ5'和Ca35'是流行最普遍的主要种群,这些种群呈现显著的地区分布特征,可以利用CRISPR分型技术加强该地区鼠疫检测和防控工作.

摘要： 目的 观察大沙鼠感染鼠疫耶尔森菌(鼠疫菌)后F1抗体反应及动态变化情况.方法 大沙鼠捕获于2011年新疆维吾尔自治区准噶尔盆地鼠疫自然疫源地南缘,共211只,其中未感染鼠疫菌的大沙鼠为167只,感染鼠疫菌的为44只.实验菌株采用2504号鼠疫菌,该株鼠疫菌硝酸盐还原实验阴性,为强毒菌.采用随机数字法选取35只未感染鼠疫菌大沙鼠并平均分为7组(6个实验组和1个对照组),用按10倍梯度稀释成的1×106~1×1011 CFU/ml浓度梯度菌液对1~6实验组大沙鼠进行第1次感染,对照组皮下鼠蹊部注射生理盐水,感染量均为1 ml;选取已感染过鼠疫菌且首次检测F1抗体滴度在1:256~1:4096之间的大沙鼠共17只,按照抗体滴度进行分为1:4096组(4只)、1:2048组(4只)、1:1024组(3只)、1:512组(3只)、1:256组(3只),并于每30天尾部无菌采血1次进行F1抗体检测,共检测5次;从剩余感染过鼠疫菌的大沙鼠中选取经2次检测F1抗体阴性的大沙鼠共9只,采用浓度为1×106 CFU/ml菌液进行第2次感染,感染量为1 ml.第1次和第2次感染后的大沙鼠,均于感染后的第3、5、7、15、30、60、90和120天尾部采血检测鼠疫F1抗体.采用一元线性回归方程建立大沙鼠抗体衰减回归模型.结果 第1次感染鼠疫菌的大沙鼠中,1×106~1×108 CFU/ml组分别在第30、15和15天时检出抗体,抗体阳性率分别为1/4、3/4、4/5,1×107和1×108 CFU/ml组均在第120天时达到最高抗体滴度,均为1:256;1×109、1×1010和1×1011 CFU/ml组,在第5天至第7天时可检出抗体,且在第7天至第15天时大沙鼠全部抗体阳转,1×1011 CFU/ml组在第120天时达到最高抗体滴度,为1:4096.第2次感染鼠疫菌的大沙鼠中,第3天即可检出抗体阳性,阳性率为2/9,至第90天达到最高抗体滴度,为1:2048.大沙鼠F1抗体衰减的一元线性回归方程为y=0.045x-0.321(F=115.40,P<0.001),从F1抗体滴度1:4096衰减至0的最短时间为140 d,最长为200 d.结论 感染高浓度鼠疫菌菌液的大沙鼠F1抗体产生的时间较短,其抗体阳性率也较高,抗体滴度最高时可达1:4096;F1抗体衰减时间长,约在140~200 d之间.%Objective To observe the dynamics of antibody response in great gerbils infected with Yersinia pestis in experiment. Method A total of 211 great gerbils were captured in the southern margin of plague natural focus of Junggar Basin of the Xinjiang Uygur Autonomous Region in 2011. Among them, there were 167 great gerbils without infection of Y. pestis and 44 great gerbils infected by Y.pestis. Y.pestis No. 2504 was employed for this experimental strain, which was strong toxic strain with negativity in the reduction experiment of nitrate. 35 great gerbils without the infection of Y. pestis were divided randomly and averagely into 7 groups including 6 experimental groups and 1 control group. Great gerbils in the 1st to 6th experimental groups were exposed first with 1 × 106-1 × 1011 CFU/ml of bacterial fluid with 10 times of gradient dilution; groin areas of great gerbils in the control group were injected subcutaneously with physiological saline;and the amount of infection was all 1 ml. 17 great gerbils infected with Y. pestis and the first detection of F1-antibody titer in 1:256-1:4096 were grouped according to F1-antibody titer: group 1:4096 (n=4), group 1:2048 (n=4), group 1:1024 (n=3), group 1:512 (n=3) and group 1:256 (n=3);and blood in caudal regions was collected in asepsis for the detection of F1-antibody, with a total of 5 times.9 great gerbils which were selected from the remaining great gerbils infected with Y. pestis and detected F1-antibody negative 2 times were exposed 1×106 CFU/ml of bacterial fluid for the second infection, with the amount of infection being 1 ml. Blood in caudal regions of great gerbils after the first and second infection were collected for the detection of plague F1-antibody on the 3rd, 5th, 7th, 15th, 30th, 60th, 90th and 120th day after infection. Declined regression models for great gerbils' antibodies were established with unary linear regression equation;declined change diagrams for the antibodies were drawn to observe the declined F1-antibody after great gerbils were exposed to Y. pestis. Results In great gerbils with the first infection of Y. pestis, antibodies were detected in the 1 × 106-1 × 108 CFU/ml of group on the 30th, 15th and 15th day, respectively; the positive rates of antibody were 1/4, 3/4 and 4/5, respectively; the group 1 × 107 and 1 × 108 CFU/ml reached to the highest antibody titer with 1:256 on the 120th day;antibodies were revealed in the group 1×109, 1×1010 and 1×1011 CFU/ml from the 5th to 7th day when the seroconversion of all antibodies was observed;group 1×1011 CFU/ml reached to the highest antibody titer on the 120th day with 1:4096. In the great gerbils with the second exposure to Y.pestis, positive antibodies were detected on the 3rd day with the positive rate being 2/9; and the highest antibody titer with 1:2048 was noted on the 90th day. Unary linear regression equation of declined F1 antibody of great gerbils was y=0.045x-0.321(F=115.40,P<0.001), and the shortest duration for F1-antibody titer declining from 1:4096 to 0 was 140 d and the longest duration 200 d. Conclusion Great gerbils infected with the high concentration of Y. pestis fluid show shorter duration in producing F1-antibody, the antibody positive rate is also higher, and the highest antibody titer can reach 1:4096. The great gerbils could hold the plague F1 antibodies for a long time which was about 140 to 200 days from the highest titer.

摘要： 目的 了解准噶尔盆地大沙鼠(Rhombomys opimus)感染鼠疫耶尔森菌(鼠疫菌)的组织病理与超微病理变化.方法 于2011年在新疆准噶尔盆地捕获40只实验大沙鼠,经实验室人工饲养6个月,且实验动物血清采用鼠疫菌F1抗体间接血凝法检测为阴性.鼠疫菌(编号:2504,属中世纪型)由新疆维吾尔自治区疾病预防控制中心2005年分离自准噶尔盆地鼠疫自然疫源地的活体大沙鼠,其毒力测定为半数致死量(LD50)<10 CFU/ml.实验共设7个实验组和1个对照组,并采用随机数字法将40只大沙鼠平均分到8个组.实验组大沙鼠分别接种按10倍梯度稀释成的7.4×105、7.4×106、7.4×107、7.4×108、7.4×109、7.4×1010和3.0×1011 CFU/ml浓度菌液,对照组接种生理盐水,接种量均为1 ml.收集所有感染鼠疫菌大沙鼠的心脏、肝脏、脾脏和肺脏组织,分别处理后进行光学显微镜镜检和透射电子显微镜观察.结果 7.4×108~3.0×1011 CFU/ml实验组均引起大沙鼠特异性感染死亡,感染动物肝脏呈点状坏死伴脂肪变性,肝细胞核内小管增多,细胞器分布不均.脾脏淋巴组织呈反应性增生,血窦腔内可见较多中性粒细胞,吞噬细胞内可见被吞噬的细菌.心肌细胞轻度变性、横纹模糊,肌细胞核基质密度降低,肌原纤维排列松散.肺脏可见血管扩张、充血,肺泡腔内呈Ⅰ型上皮细胞脱落.大沙鼠感染7.4×105~7.4×107 CFU/ml鼠疫菌后未出现特异性感染死亡,实质器官的组织病理与超微病理相较于7.4×108~3.0×1011 CFU/ml组改变轻微.结论 鼠疫菌可以引起大沙鼠实质器官的组织病理和超微病理发生改变;肝脏和脾脏可能是鼠疫菌感染大沙鼠的主要靶器官.%Objective To understand the histopathological and ultrastructural pathology changes of great gerbils in the Junggar Basin to Yersinia pestis infection. Methods Forty captured great gerbils from the Junggar Basin that tested negative for anti-F1 antibodies were infected. The Y. pestis strain 2504, isolated from a live great gerbil in the natural plague foci of the Junggar Basin in 2005 with a median lethal dose (LD50) of<10 CFU/ml, was used in this study. Forty great gerbils were divided into seven infection groups and were subcutaneously infected with 7.4×105, 7.4×106, 7.4×107, 7.4×108, 7.4×109, 7.4×1010, or 3.0×1011 CFU/ml of 2504. One milliliter of physiological saline was injected in the noninfected group as a control. We collected the liver, spleen, heart, and lung from all animals for histopathologic and ultrastructural pathology examination. Results Great gerbils in the 7.4 × 108-3.0 × 1011 CFU/ml groups did not survive and exhibited pathological changes and altered ultrastructural pathology. The liver tissue of infected great gerbils showed spotty necrosis and fatty degeneration, intranuclear canaliculi with increased hepatocytes, and uneven distribution of organelles. Additionally, reactive proliferation of lymphoid tissue in the spleen, blood sinusoid lacunae with neutrophil infiltration, and phagocytosed bacteria in phagocyte cells were observed. Myocardial fiber hypertrophy and interstitial indistinction, nuclear matrices decreased in cardiac myocytes, and loose arrangement of myogenic fibers in myocardial cells were also observed. Angiectasia, capillary congestion, and tissue necrosis were found in the lung. No significant difference in histopathological and ultrastructural pathology in the parenchymal organ was observed between the 7.4×105-7.4×107 CFU/ml groups and the 7.4× 108-3.0×1011 CFU/ml groups, and no specific death caused by Y. pestis infection was apparent in the 7.4×105-7.4 × 107 CFU/ml groups. Conclusion Y. pestis infection altered tissue and ultrastructural pathology in the parenchyma apparatus of great gerbils. In particular, the liver and spleen appeared to be the primary site of Y. pestis infection in great gerbils.

摘要： 本文描述了用于表达鼠疫耶尔森氏菌F1‑V融合蛋白的质粒构建体的产生。在所公开的质粒中，所述F1‑V融合蛋白编码序列与大肠杆菌htrA启动子可操作地连接，并且与大肠杆菌HlyA分泌信号序列框内融合。还描述了包含所述F1‑V融合蛋白表达质粒的肠道沙门氏菌血清变型伤寒沙门氏菌菌株Ty21a，例如用作抗鼠疫的口服疫苗。

摘要： 本发明涉及一种检测炭疽杆菌、鼠疫耶尔森菌及嗜肺军团菌多重实时荧光PCR的引物、探针序列及方法，该引物、探针序列包括炭疽杆菌、鼠疫耶尔森菌及嗜肺军团菌引物探针序列集；该方法的步骤包括选取引物对和探针、制备目标基因的质粒标准品、建立反应体系和选择荧光检测通道。本发明设计的各引物对和探针特异性和工作性能良好，可建立检测炭疽杆菌、鼠疫耶尔森菌及嗜肺军团菌的多重实时荧光PCR方法，具有结果可靠和准确灵敏、操作简便、省时省力、成本低、效率高等优点。

摘要： 人源化抗鼠疫耶尔森菌抗原F1的抗体及应用，所述抗体的，轻链可变区具有SEQ ID NO:1核苷酸序列和SEQ ID NO:2氨基酸序列，其中轻链超变区序列具有SEQ ID NO:3、SEQ ID NO:4和SEQ ID NO:5；重链可变区具有SEQ ID NO:6核苷酸序列和SEQ ID NO:7氨基酸序列，重链超变区具有SEQ ID NO:8、SEQ ID NO:9和SEQ ID NO:10。该抗体能够特异性结合鼠疫耶尔森菌抗原（F1）。

摘要： 本发明公开了一种基于实时荧光RPA技术的鼠疫耶尔森菌检测试剂盒及其应用，所述试剂盒成套试剂，由引物对和名称为YPS723‑1‑P1的探针组成，所述引物对由名称分别为YPS723‑F1和YPS723‑R1的单链DNA组成。本发明的试剂盒能够快速、便携、灵敏度高、特异性好的检测鼠疫耶尔森菌，有望成为临床样本的快速诊断辅助工具。

摘要： 本发明提供了一种检测鼠疫耶尔森氏菌的试剂，它包括上游引物、下游引物、荧光探针和淬灭探针。本发明还提供了利用上述试剂荧光定量PCR检测鼠疫耶尔森氏菌的方法。本发明根据鼠疫耶尔森氏菌6MD质粒上特有的致病基因-血浆凝固酶及胞浆素原活化因子pla基因，设计引物和探针，与整个genebank数据库进行BLAST检索，发现其只对鼠疫菌上的pla基因序列特异，和其它物种的核酸序列无同源，保证了检测方法的特异性。该方法可在同一管内实现DNA的扩增和同步检测，而无需在PCR后进行凝胶电泳分析，对样本的定量检测只需2小时左右即可完成，具有操作简便，高效、快速、特异的优点。

摘要： 本文描述了用于表达鼠疫耶尔森氏菌F1-V融合蛋白的质粒构建体的产生。在所公开的质粒中，所述F1-V融合蛋白编码序列与大肠杆菌htrA启动子可操作地连接，并且与大肠杆菌HlyA分泌信号序列框内融合。还描述了包含所述F1-V融合蛋白表达质粒的肠道沙门氏菌血清变型伤寒沙门氏菌菌株Ty21a，例如用作抗鼠疫的口服疫苗。

摘要： 本发明提供了一种鼠疫耶尔森氏菌菌蜕、其制备方法及作为疫苗的应用。所述菌蜕具有完整的外膜结构，无需佐剂接种小鼠即能够诱导机体产生保护性抗体，产生较强的免疫记忆，可保护小鼠免受强剂量鼠疫活菌株的攻击，与活疫苗相比具有更高的安全性。

摘要： 本发明涉及一种检测炭疽杆菌、鼠疫耶尔森菌及嗜肺军团菌多重实时荧光PCR的引物、探针序列及方法，该引物、探针序列包括炭疽杆菌、鼠疫耶尔森菌及嗜肺军团菌引物探针序列集；该方法的步骤包括选取引物对和探针、制备目标基因的质粒标准品、建立反应体系和选择荧光检测通道。本发明涉及的各引物对和探针特异性和工作性能良好，可建立检测炭疽杆菌、鼠疫耶尔森菌及嗜肺军团菌的多重实时荧光PCR方法，具有结果可靠和准确灵敏、操作简便、省时省力、成本低、效率高等优点。

摘要： 本发明公开了一种快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的检测方法，利用基质辅助激光解析电离飞行时间质谱检测小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌的肽指纹图谱，根据两种耶尔森氏菌特征峰的不同，鉴定其特征蛋白，区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌。本发明采用的方法比现有的传统生化实验鉴定时间缩短1‑2天，可以快速区分小肠结肠炎耶尔森氏菌与中间型耶尔森氏菌，实现快速检测。该方法自动化程度高，检测时间短，有效降低一线检测人员接触食源致病菌的几率，减少食源致病菌对人和环境污染的风险。

摘要： 本发明提供了一种选择敏感培养基对腐败材料检测鼠疫耶尔森氏菌检测的应用效果，属于鼠疫菌分离培养技术领域。本发明提供的从腐败材料中分离培养鼠疫耶尔森菌的培养基，包括如下原料：0.1％溶血赫氏培养基、十二烷基硫酸钠、红霉素、胆盐、阿莫西林、甘露醇和中性红。本发明培养基中性红指示剂在酸性环境中产生红色集团附着于菌落上，使鼠疫菌菌落显示红色，对变形杆菌、大肠杆菌和革兰氏阳性菌抑制作用强，适用于检验腐败和杂菌污染的动物材料，具有高特异性和强敏感性，培养24h鼠疫菌发育为典型成熟菌落。而且生长的鼠疫菌菌落典型，易于挑选，在野外材料鼠疫菌分离有实际意义。