贵州省2株牛种布鲁氏菌分子流行病学特征分析

目的 分析贵州省牛种布鲁氏菌分子流行病学特征,为牛种布鲁氏菌病疫情的防控提供科学依据。方法 采用BCSP31-PCR及AMOS-PCR技术对经传统方法鉴定为牛种布鲁氏菌的2株分离株进行属/种复核,采用基于布鲁氏菌rpoB基因的单核苷酸多态性分析了解其rpoB基因型,采用MLVA-16技术进行MLVA分型,了解其与国内牛种布鲁氏菌的聚类关系,采用MLST技术分析其序列型及其与国内菌株间的遗传进化关系。结果 2株分离株经BCSP31-PCR和AMOS-PCR技术被鉴定为布鲁氏菌属细菌,未能明确其具体种型;其rpoB基因型相同;MLVA分型与新疆牛种布鲁氏菌共享同一MLVA-16型(4-5-3-12-2-2-3-1-6-43-8-5-5-5-3-3); MLST分析鉴定2株菌均为ST2型,最小间距图显示2株菌与布鲁氏菌属内的牛种布鲁氏菌遗传距离最近,属同一个遗传复合体。结论 贵州省2株牛种布鲁氏菌分离株间的遗传距离近,与新疆牛种3型布鲁氏菌为同一MLVA型,提示分离株引起的感染可能为外省输入。     

福建省布鲁氏菌分离株的分子生物学鉴定

目的利用分子生物学方法鉴定福建省布鲁氏菌分离株。方法对布鲁氏菌分离株进行AMOS-PCR和MLVA分型。结果3株福建省分离株经AMOS-PCR鉴定为羊种,与传统生物学分型一致;MLVA分型将其分为3个基因型,聚类分析鉴定为羊种。结论AMOS-PCR、MLVA分型可以作为我省布鲁氏菌分离株鉴定的辅助手段。     

Genetic analysis of mycobacterium tuberculosis strains isolated in Ural region, Russian Federation, by MIRU-VNTR genotyping.

SETTING: The Ural region in Russia is one of the areas most affected by a high incidence of tuberculosis (TB). Molecular epidemiological studies able to trace Mycobacterium tuberculosis transmission are of particular significance. OBJECTIVE: To characterize the population of M. tuberculosis strains circulating in the Ural region, to detect the predominant genotypes and to evaluate their phylogenetic relationship and epidemiological significance. DESIGN: Ninety-two M. tuberculosis clinical samples originating from the Ural region were genotyped using the MIRU-VNTR method. RESULTS: Two major phylogenetically distinct groups of isolates were identified: the W-Beijing family (54.3%) and a previously unreported cluster, named the Ural group (15.2%). Forty-seven different MIRU profiles were identified, including 38 unique (41.3%) and 54 isolates grouped into nine clusters (from 2 to 28 isolates in each cluster). Genetic diversity within the clusters was shown by additional sub-typing of M. tuberculosis isolates in nine additional QUB-VNTR loci. CONCLUSION: W-Beijing family isolates are associated with multiresistance to anti-tuberculosis drugs. It is possible that the strains of this family play a significant role in the spread of multidrug-resistant TB over the Ural region.     

Anthropozoonotic role of Brucella abortus

During a severe outbreak of brucellosis in buffaloes, two animal attendants and one Veterinarian contracted Brucella abortus infection. All the three persons developed clinical signs of brucellosis. A high antibody titre against Br. abortus was demonstrated in the sera of these sick individuals. The organism could be isolated from the blood of one patient on tryptose agar at 37 degrees C. Clinical spectrum, chemotherapy and prevention of brucellosis has been discussed.     

福建省首次从人体分离到的布鲁氏菌的初步研究

目的对从人体分离到的5株布鲁氏菌进行种型鉴定。方法常规确定布鲁氏菌属与种型鉴定。结果FJFZ0609菌株由于菌型特殊,难以定种;FJZZ0701、FJNP0808和FJSM0808三个菌株均为羊种布鲁氏菌2生物型;FJFZ0903菌株为羊种布鲁氏菌3生物型。结论福建省自1964年以后首次从人体内分离到羊种布鲁氏菌,证实福建省存在羊种布鲁氏菌并波及人间。     

内蒙古布鲁氏菌临床分离株多位点序列分型研究

目的探讨临床分离布鲁氏菌的种群结构和遗传进化特征。方法布鲁氏菌标准参考菌株羊种16M,牛种544A和猪种1 330 S作为对照菌株,采用多位点序列分型(Multiple locus sequence typing,MLST)方法对内蒙古地区分离的116株羊种布鲁氏菌进行分析,确定待测菌株的序列型(STs),用软件BioNumerics(Version 5.0)构建菌株的最小生成树。结果116株羊种布鲁氏菌全部为ST8型,9个MLST位点的变异完全相同,分离株种群结构单一;羊种菌的生物型与ST型无明显关联,ST8型菌系该地区的主要流行菌种,并与内蒙古地区先前分离羊种布鲁氏菌进化程度相同,且有密切的亲缘关系。结论羊种布鲁氏菌的序列分型(ST)与常规生物分型存在差异,ST8型菌可能具有较强的环境适应性和致病性。MLST是一种较好的羊布鲁氏菌种群和进化关系研究方法,但更适合于具有较大时间跨度和地理分布菌株间的流行病学调查。     

Detection of Brucella melitensis and Brucella abortus by DNA amplification.

Suitable reaction conditions and oligonucleotide primers were sought for the detection of Brucella melitensis and Brucella abortus by the polymerase chain reaction. Primers were chosen from within the coding sequence of a gene encoding a 31 kDa B. abortus antigen. The test was shown to be sensitive, and specificity was demonstrated using DNA derived from a panel of Gram-negative pathogens. There was no detectable difference between B. melitensis and B. abortus in the sensitivity of the reaction or in the size of the amplification product. The technique should be applicable in the diagnosis of brucellosis.     

Current ascariasis situation in the Russian Federation

In the Russian Federation (RF), ascariasis remains to be the most common geohelminthiasis, in children in particular. With its overall reduction, the prevalence of ascariasis is very uneven on the RF territory. In the past 15 years, from 1991 to 2004, there was a steady-state decrease in the incidence of ascariasis from 79.5 to 46.0 per 100,000. Its incidence rates remained higher than the average Russian ones in 33 RF subjects and lower than those in 48 subjects and began reducing in 8 subjects in the past 4 years. Of the 33 RF subjects wherein the incidence rates were constantly greater than the average Russian ones, 27 subjects are located in the natural background-favorable areas. In the past 15 years, there was an increase in the impact of socioeconomic factors on the higher incidence of ascariasis: the population's migration; the delivery and transport of Ascaris egg-contaminated greens, vegetables, berries; the development of hothouse gardening, and, as a result, the occurrence of ascariasis in the northern areas of the country, as well as the use of discharged waters for soil fertilization to grow crops. The territory of Russia was first epidemiologically districted in the context of ascariasis, by taking into account the natural background and socioeconomic factors. The area of ascariasis is on the rise. In all RF subjects, public health and social security bodies should constantly keep in mind the problem in ascariasis control and prevention.     

Contraindications to vaccination in the Russian Federation

Failure to achieve high levels of immunity among children contributed to the epidemic of diphtheria that occurred in the Russian Federation during the 1990s. A major factor in this failure was the extensive list of contraindications to vaccination that was in use throughout the countries of the former Soviet Union. In 1980, the Ministry of Health (MOH) of the Soviet Union adopted an extensive list of contraindications for use of the diphtheria-tetanus toxoids-pertussis (DTP) vaccine. In 1994, the MOH of the Russian Federation revised the list of contraindications to vaccination to be largely in accord with World Health Organization recommendations. Since then, age-appropriate vaccination coverage has increased markedly: In 1996, DTP3 coverage among children 12 months of age had increased to 87% from 60% in 1990.     

快速检测牛羊猪种布鲁杆菌多重PCR的建立

目的 建立一种能同时快速检测并能鉴别牛、羊、猪种布鲁杆菌的多重PCR方法.方法 根据IS711插入序列设计1条公共引物和3条牛、羊、猪种布鲁杆菌(544A、16M、1330S)特有序列引物,进行多重PCR反应;选择耶尔森菌O:9、大肠埃希菌O157:H7、鼠伤寒沙门菌47729进行多重PCR反应的特异性检测;倍比稀释定量法观察牛种布鲁杆菌多重PCR反应的敏感性.结果 牛、羊、猪种布鲁杆菌多重PCR反应扩增片段产物长度分别为485、731、248 bp;耶尔森菌O:9、大肠埃希菌O157:H7、鼠伤寒沙门菌47729加入布鲁杆菌中进行多重PCR反应.扩增结果呈阴性;牛种布鲁杆菌多重PCR反应敏感性为0.0967 Pg.结论 成功建立快速检测牛、羊、猪种布鲁杆菌多重PCR扩增反应方法,且其特异性、敏感性较好.     

1952～2003年陕西省检出布鲁氏菌特性分析

目的 总结陕西省1952～2003年的布鲁氏菌检出结果，系统分析布鲁氏菌病的流行状况及病原学研究进展。方法 把全省历年布鲁氏菌病检菌资料分为1952～1981、1982～1991和1992～2003年3个阶段进行统计分析。结果 52年来，陕西省从8个地市5种宿主的不同材料中共分离出3种228株布鲁氏菌，其中羊种菌222株，牛种和犬种菌各3株；自人、羊、鹿、犬、牛分离到的菌株数分别为185、24、15、3和1；1991年以前分离的羊种菌为1、2型，而1996年以后则为羊1、3型。对15株羊种菌进行毒力测定，均为强毒株。结论 陕西省是以羊种为主的羊、牛、犬种布鲁氏菌病混合疫区。流行菌株毒力强，有严重的致病性。陕北、关中疫情严重，陕南存在犬种布鲁氏菌病疫源地。发现羊种菌有宿主转移现象。     

Diphtheria in the Russian Federation in the 1990s

A resurgence of diphtheria spread throughout the Russian Federation in the early 1990s; diphtheria had been well controlled, but circulation of toxigenic strains of Corynebacterium diphtheriae had persisted since the implementation of universal childhood vaccination in the late 1950s. More than 115,000 cases and 3,000 deaths were reported from 1990 to 1997, and, in contrast to the situation in the prevaccine era, most of the cases and deaths occurred among adults. Contributing factors included the accumulation of susceptible individuals among both adults and children and probably the introduction of new strains of C. diphtheriae. Vaccine quality, vaccine supply, or access to vaccine providers did not significantly contribute to the epidemic. Mass vaccination of adults and improved childhood immunization controlled the epidemic. High levels of population immunity, especially among children, will be needed to prevent and control similar outbreaks in the future.     

多位点序列分型在新疆人间布鲁氏菌病临床分离株遗传进化研究中的应用

目的 探讨多位点序列分型(MLST)技术在新疆人间布鲁氏菌病分离株遗传进化研究中的应用价值,了解分离株的种群结构和遗传进化关系。方法 采用MLST对2015、2016年分离自新疆7个地州的24株人间布鲁氏菌病分离株和3株布鲁氏菌标准参考菌株进行分析,统计各菌株序列型(STs),运用BioNumerics软件构建菌株最小进化树,分析菌株间遗传关系;收集过往研究的186株全国不同地区布鲁氏菌分离株MLST结果,分析各ST型分布特点。结果 24株人间布鲁氏菌分离株全部为ST8型,3株标准参考菌株(羊种16M、牛种544A、猪种1330S)分别为ST7型、ST1型、ST14型;24株人间布鲁氏菌分离株的9个MLST位点变异完全相同,分离株种群结构单一。ST8型菌是我国主要流行的布鲁氏菌,且以北方流行为主;MLST能较好的区分布鲁氏菌种别,但不能有效区分生物型别。结论 ST8型菌(羊种3型)是新疆人间布鲁氏菌病的主要流行菌株,MLST技术可作为人间布鲁氏菌种群结构和遗传进化关系研究的补充手段。     

Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR

ObjectiveTo evaluate simultaneous detection and differentiates of Brucella abortus (B. abortus) and Brucella melitensis (B. melitensis) through the combinatorial PCR method.MethodsThis study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals. Identification and differentiation of each species using the size of the PCR product were determined. To determine the specificity of the method, bacteria close to the genus Brucella were used. Finally, to confirm PCR products, In addition to the products sequence, RFLP was performed on PCR products using restriction enzymes.ResultsThe method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B. abortus and B. melitensis with high specificity and sensitivity in clinical samples. Differentiation of species is based on the resulting bands; therefore, the band 494 bp for B. abortus and 733 bp for B. melitensis were obtained. RFLP and sequencing results confirmed PCR results.ConclusionsThe results of this study shows that without routine diagnostic methods such as culture and serology tests, using the molecular method of combinatorial PCR, important species of Brucella can be simultaneously identified and differentiated in clinical samples.     

Molecular detection of Brucella abortus in wild and captive felids

PurposeBrucellosis is a zoonotic disease of great public health importance. In wild animals, Brucella abortus is one of the most diagnosed species, mainly in enzootic environments where domestic animals share the same environment. B. abortus is common in environments shared by cattle, wild, and domestic animals. This study aimed to detect the presence of B. abortus DNA in free-ranging and captivity felids at Mato Grosso State, Brazil.MethodPolymerase chain reaction, based on the genetic element IS711, was performed in blood samples collected from 23 free-ranging and captive felids. The species represented include Leopardus colocolo, Leopardus pardalis, Leopardus wiedii, Panthera onca, Puma concolor, and Puma yagouaroundi.ResultsDNA amplification of B. abortus was observed in only one captive P. concolor (4.34%).ConclusionThe detection of this pathogen in captive animals using molecular tools demonstrates the importance of monitoring, as it raises concerns about the possibility of transmission between humans and wild and domestic animals, especially in regions of vast biodiversity, such as in the State of Mato Grosso, Brazil.