16SrRNA基因序列分析用于诊断病原性细菌感染初步研究

目的建立一种可适用于多种病原菌检测的方法,以提高病原学诊断率,有利于传染病的病原学监测。方法细菌16SrRNA基因通用引物对直接从血液、尿液、脑脊液标本中提取的细菌DNA进行PCR扩增,并对16SrRNA基因扩增产物进行克隆和序列分析。结果检测了28份临床标本,16SrRNA基因序列分析与尿液细菌培养结果不完全吻合;血液和脑脊液的16SrRNA的检出率均高于细菌培养。结论16SrRNA基因检测分析和细菌培养均具有良好的诊断效果,而针对16SrRNA基因的PCR直接扩增核酸并进行序列分析可对分离培养阴性的标本进行有效的补充诊断,可检测到多种病原菌如肠球菌,大肠杆菌,表皮葡萄球菌,牛链球菌,猪链球菌,布鲁氏菌,表皮葡萄球菌和脑膜炎球菌等,为疾病的诊断提供良好的线索。     

人胃螺杆菌16SrRNA基因序列测定

目的 从分子水平探讨胃螺杆菌（Ｈｅｌｉｃｏｂａｃｔｅｒ ｈｅｉｌｍａｎｎｉｉ，Ｈｈ）的生物分类学地位。方法 采用螺杆菌属特异和细菌通用的１６ＳｒＤＮＡ引起对１例Ｈｈ阳性胃粘膜组织行ＰＣＲ扩增、产物纯化和四荧光标记单泳道法自动测序，ＰＣ／ＧＥＮＥ软件分析其与各近缘菌之间的同源性。结果 与国外所侧两例Ｈｈ的同源笥分别为９８．４％和９６．０％，与螺杆菌属细菌的的同源笥均在９３．５％以上，与猫胃螺杆菌（Ｈ     

应用16SrRNA基因序列分析技术鉴定临床非典型细菌的研究

目的将16SrRNA基因序列分析技术应用于传统方法鉴定可靠率低或无法鉴定细菌的分类鉴定,以提高临床诊断的准确性。方法以本院微生物实验室保存的5株标准菌株验证实验方法的准确性后,收集实验事常规方法不能鉴定或鉴定可靠性低的临床分离菌15株（包括革兰阳性球菌,革兰阳性杆菌及革兰阴性杆菌）,提取DNA模板,以通用引物PCR扩增16SrRNA目的片段后测序,将测序结果在GenBank数据库中比对分析以确定菌种。结果15株实验细菌中有14株（占93．3％）鉴定到种,包括鼻疽诺卡菌,大肠埃希菌,阿尔莱特葡萄球菌,脆弱拟杆菌,弗氏柠檬酸杆菌,短小芽孢杆菌,河流漫游球菌,极小短小杆菌,伴放线凝聚杆菌,毗邻颗粒球菌;1株菌鉴定到属（占6．7％）,归属于垃状杆菌属。结论16SrRNA基因序列分析技术具有准确、快速和方法简便的特点,适用于对临床非典型蔚、少见菌以及新型细菌的鉴定,可作为细菌常规鉴定手段的必要补充。     

云南省曲靖市啮齿动物的立克次体血清学和分子流行病学调查

目的调查云南省曲靖市啮齿动物感染立克次体状况。方法用鼠笼法捕鼠,采集鼠血清和脾脏。用间接免疫荧光试验(IFA)检测鼠血清中7种常见立克次体的IgG抗体;用巢氏PCR方法检测鼠脾中嗜单核细胞埃立克体和嗜吞噬细胞无形体16SrRNA基因片段部份序列。结果 2012年7-9月在曲靖市捕获啮齿动物3种592只,褐家鼠(Rattus norvegicus)和黄胸鼠(Rattus flavipectus)构成比分别为61.49%和35.47%。鼠血清中贝氏柯克斯体(Coxiella burentii)、莫氏立克次体(Rickettsia typhi)、恙虫病东方体(Orientia tsutsugamushi)、西伯利亚立克次体(Rickettsia sibirica)、嗜吞噬细胞无形体(Anaplasma phagocytophilum)和嗜单核细胞埃立克体(Ehrlichia chaffeensis)的抗体阳性率依次为21.45%、7.60%、7.09%、3.38%、1.18%和0.51%;未检测到猫立克次体(Rickettsia felis)抗体。从5份褐家鼠脾脏标本中检测到16SrRNA基因序列,同源性和进化分析表明,1株为嗜吞噬细胞无形体,4株为莫氏立克次体。结论曲靖市啮齿动物中存在莫氏立克次体和人粒细胞无形体流行,褐家鼠可能是主要宿主;同时还存在恙虫病东方体、贝氏柯克斯体、斑点热立克次体和埃立克体的感染。当地疾控和医疗机构应加强立克次体病的监测。     

定量PCR联合基因芯片检测腹水细菌16SrRNA诊断自发性细菌性腹膜炎

目的 探讨定量PCR联合基因芯片检测腹水细菌16SrRNA基因诊断自发性细菌性腹膜炎(SBP)的意义.方法 采用实时荧光定量PCR联合基因芯片检测76例临床疑似SBP肝病患者和6例对照的非感染性腹腔积液肝病患者腹水细菌16SrRNA基因,与腹水细菌培养同时比较.结果76份疑似SBP患者腹水标本中,定量PCR联合基因芯片检测阳性17份,阳性率为22.4%,其中革兰氏阳性菌8份、革兰氏阴性菌9份;腹水细菌培养阳性6份,阳性率为7.9%,均为革兰氏阴性菌,两种方法比较,x2=18.05,P＜0.01,差异有统计学意义.两种方法检测腹水细菌阳性的6份标本,菌株鉴定结果相一致.对照病例细菌检测结果呈阴性.结论 定量PCR联合基因芯片检测腹水细菌16SrRNA基因,较腹水细菌培养的敏感性和特异性高;不仅能作出快速诊断,还能确定SBP所感染的病原菌,具有实际应用价值.

Abstract：

Objective To evaluate the significance of determining ascitic bacterial16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP). Methods Ascitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases.The results were compared with ascitic bacterial culture simultaneously. Results Of 76 ascitic samples, 17were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative (G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P ＜ 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of noninfectious ascites with chronic liver diseases. Conclusions Determination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.     

新疆伊犁地区无形体流行病学调查及病原16SrRNA序列分析

目的调查新疆地区人、家畜动物包括羊、牛、马及野生动物旱獭、大尾黄鼠及家养马鹿等无形体感染及病原16SrRNA序列特征。方法采集伊犁地区正常人、牛、马、羊及野生啮齿动物大尾黄鼠、旱獭血液。 微量间接免疫荧光检测方法（mIFA）检测血清无形体IgM、IgG抗体。巢氏PCR扩增无形体16SrRNA基因并分析序列特征。结果当地人群无形体IgG抗体阳性率为5.3%;羊、牛及马为6.9%、6.3%及9.1%。 PCR结果发现羊、牛及马无形体16SrRNA基因检出率为38.9%、37.5%及36.4%。16SrRNA（228bp）序列分析显示存在明显变异株,同源比较结果提示这些变异株在我国其他地区及周边国家媒介及动物宿主中均有广泛分布。结论新疆伊犁地区存在人及家畜无形体感染。 开展发热病人无形体实验室调查及进一步人群、媒介种类、宿主与无形体生态学关系研究十分必要。     

用16SrRNA基因诊断自发性腹膜炎23例

１材料和方法１．１材料２３例肝硬变失代偿期患者,男１７例,女６例,平均年龄４２６岁,全部为近期河南医科大学一附院消化科住院的患者,均有中等量或大量腹水,利尿治疗效果不佳．１．２标本腹穿取腹水１０ｍＬ,６０００ｒ／ｍｉｎ离心１０ｍｉｎ～１５ｍｉｎ,沉...     

立克次体目微生物的系统分类进展

立克次体目（Rickettsiales）微生物是一类严格细胞内寄生的原核细胞型微生物。其生物学性状如形态结构、化学组成及代谢方式等方面与细菌类似。立克次体是引起斑疹伤寒、恙虫病、Q热等传染病的病原体,首先由美国青年医师Howard Taylor Ricketts发现,为纪念他在研究斑疹伤寒时不幸感染而献身,故以他的姓名命名。立克次体与节肢动物关系密切,寄生在吸血节肢动物体内,使其成为寄生宿主,或为储存宿主,或同时为传播媒介。立克次体大多是人兽共患病的病原体,引起的新现和再现传染病（Emergingand re-e—merging infectious diseases）呈世界性或地方性流行,日益受到医学界重视。近年来,随着立克次体分子生物学研究的迅速发展（如16SrDNA序列分析、全DNA或基因片段分析等）,原有的立克次体分类已不能准确反映立克次体目、科和各种属的面貌,代之而起的是根据特定基因（如16SrRNA基因、HSP60基因等）对立克次体进行新的分类。     

食酸代尔夫特菌16SrRNA检测及生物学特性研究

目的监测食酸代尔夫特菌的生物学特征、分子生物学特性及其与感染的关系。方法血标本培养阳性者,革兰染色阴性杆菌、球杆菌,进行形态、培养特征、细胞壁化学成分分析和16SrRNA 基因测序。结果经形态学、培养特征和16SrRNA 基因测序,鉴定为食酸代尔夫特菌。测定的1 418 bp序列与此种菌的模式株的序列100%一致,分离菌株与模式株的生化反应有一项不同。结论本文报道食酸代尔夫特菌的53种生物学特性,为常规生化表型鉴定提供借鉴。     

山西恙虫病立克次体分型及其56—kD蛋白基因序列分析

作者在我国新发现的山西恙虫病疫区病人血液中分离出3株恙虫病立克次体，SXh951，Sxh952，Sxh953。为明确其分类地位．对其进行了血清型，基因型和56－kD蛋白基因的部分序列分析。用免疫荧光抗体染色法将病人血清抗体与Karp,Gilliam．Kato株抗原反应．结果显示山西株与Gilliam型一致；用来自56－kD蛋白基因的引物扩增后，分别用限制性核酸内切酶Hinfl，HaeⅢ．Hhal消化DNA片段的多态性（PCR／RFLP）显示山西3株基因型一致，但均与Karp，Kato，Gilliam株的电泳图谱不同，显示为一独有的PCR／RFLP基因型；对Sxh951株56-kD蛋白基因片段扩增产物（1．2kb）进行DNA序列分析后．根据所推导氨基酸序列，比较Sxh951株与三个标准株氨基酸序列的同源性，发现Sxh951株与Karp、Kato和Gilliam株均有明显差异。对此，本实验室正在进一步研究，以便进一步确定其分类地位。     

肠球菌为主要菌群的腹泻标本的16S rRNA与PFGE分析

目的了解肠球菌为主要菌群的腹泻标本的微生态及分离肠球菌菌株的克隆特征。方法根据细菌16S rRNA保守区序列设计通用引物,以腹泻病人粪便标本中的总DNA为模板,PCR扩增16S rRNA片段。将获得的片段克隆入T载体进行测序,与数据库中发表的序列进行比对,判断标本中细菌的种类和比例,并对每份标本分离的各50株肠球菌选择20株进行PFGE分析。结果对4份标本的多次16S rRNA序列分析表明,该4份标本均以Enterococcus faecium为主(所占比例>68%),PFGE结果显示每个病人分离屎肠球菌菌株是克隆性的(一份标本除外)。结论从16S rRNA和PFGE的角度对腹泻粪便标本进行了分析,得到腹泻儿童肠球菌分布的基本特征,其致病性和相关机理还待今后实验范围的进一步扩大和研究的深入。     

分支杆菌临床分离株的16S rRNA基因序列分析

目的　建立分支杆菌的 16SrDNA序列分析的方法 ,采用该方法鉴定临床结核分支杆菌分离株。方法　用PCR从重庆某医院的肺部感染病人的痰标本中分得的 2 9株分支杆菌菌株中扩增 16SrRNA基因 (16SrDNA)的 5′端片段 (～5 0 0bp) ,并测定所得到片段的DNA序列。用DNA分析软件将所获得的序列与GenBank的分支杆菌序列相比较 ,计算种间相似性 ,并用序列构建分支杆菌菌株的系统发育树 ,对分离株进行分类与鉴定。结果　有 14株的 16SrDNA序列分别与已知结核分支杆菌复合群 (Mycobacteriatuberculosiscomplex ,MTC)、戈登分支杆菌、新金色分支杆菌、氯酚红分支杆菌、龟分支杆菌或脓肿分支杆菌的序列完全相同。依据完全一致的序列可以准确鉴定这些临床分离株为相应的种 ;部分分离株的 16SrDNA在GenBank序列检索中无完全一致的匹配序列。用临床分离株的 16SrDNA序列与已知的分支杆菌 16SrDNA序列构建的系统发育树 ,结果提示某些菌株可能是与已知分支杆菌密切相关的新种或是它们的亚种。结论　 16SrDNA序列分析鉴定分支杆菌是一种快速、可靠、成本较低的方法 ;重庆地区非结核分支杆菌感染的种类与其它地区的报告有明显不同     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.     

4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.