Tick-borne pathogens pose a significant threat to human health. In this study, a multiple droplet digital PCR (ddPCR) assay was developed to detect four tick-borne pathogens: Borrelia burgdorferi sensu lato (Bbsl), Coxiella burnetii (C. burnetii), spotted fever group Rickettsia (SFGR), and Borrelia miyamotoi (B. miyamotoi). Based on the singleplex ddPCR reaction system of Bbsl, the primer probes of the other three species were incorporated to develop a multiplex ddPCR reaction system. The annealing temperature and the final concentration of the primer probes were then optimized for multiplex ddPCR. The multiplex ddPCR assay was assessed for its sensitivity, specificity, repeatability, and ability to detect simulated and actual samples. The developed multiplex ddPCR approach enables the simultaneous detection of Bbsl, C. burnetii, SFGR, and B. miyamotoi. The positive target microtitre clusters are closely grouped and distinctly separated from each other, with the multiplex ddPCR assay demonstrating a dynamic range of five orders of magnitude. The limits of detection (LOD) for the multiplex ddPCR assay were 4 copies/20 µL for Bbsl, 3 copies/20 µL for C. burnetii, 3 copies/20 µL for SFGR, and 2 copies/20 µL for B. miyamotoi. The assay demonstrated high specificity, with no observed cross-reactivity against non-target pathogens. Performance was validated using both spiked samples and field-collected clinical specimens. In the evaluation of 30 ticks and 30 serum samples, the ddPCR method (in both singleplex and multiplex formats) achieved higher positive detection rates for all four target pathogens compared to quantitative real-time PCR (qPCR). In addition, the detection proportions of multiplex and singleplex ddPCR were consistent. Multiplex ddPCR can detect low DNA concentrations in samples and enables the absolute quantification of Bbsl, C. burnetii, SFGR, and B. miyamotoi, providing a novel detection approach for the clinical diagnosis of tick-borne diseases.

Objective: Fleas are vectors for the transmission of various pathogens. The study is to understand the pathogens carried by parasitic fleas in domestic dogs and to evaluate the pathogenic potential risk to humans. Methods: 121 fleas were collected from 6 dogs in different farmers' households in Suichuan County, Ji'an City, Jiangxi Province in July 2023. Flea species were determined through morphological identification and CoII gene detection. Whole genomic DNA was extracted from all 121 fleas, and six pathogens - Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, Coxiella burnetii, spotted fever group Rickettsia, and Ehrlichia chaffiensis - were detected using nested polymerase chain reaction (PCR). Results: Positive products were sequenced, and the carrier status of each pathogen was analyzed. Of the 121 fleas identified, 118 were Pulex irritans and 3 were Ctenocephalides felis. PCR results revealed that Borrelia burgdorferi (5%, 6/118), Borrelia miyamotoi (0.8%, 1/118), Borrelia hermsii (9%, 11/118), Anaplasma phagocytophilum (0.8%, 1/118), spotted fever group Rickettsia (0.8%, 1/118), and Coxiella burnetii (0.8%, 1/118) were detected in Pulex irritans. Additionally, one sample showed mixed infection with both Borrelia burgdorferi and Anaplasma phagocytophilum (0.8%, 1/118). Conclusions: This study suggests that Pulex irritans can carry multiple pathogens, with implications for public health needs that warrant further investigation.

Objective: Distribution of the bacterium flora in ticks has an essential role in mapping and preventing local tick-borne diseases. The purpose of the current study was to map the bacterium flora and identify the main pathogens in ticks in grazing areas of Yunnan province. Methods: The V3-V4 region of 16S rRNA amplifier sequencing was used to analyze the tick-borne pathogens in Rh. Microplus . A prevalence survey on B. burgdorferi s.l. , B. miyamotoi , E. chaffeensis , A. phagocytophilum , Coxiella burnetiid , and the spotted fever group rickettsiae was carried out using PCR. Phylogenetic analysis was used to identify and cluster the main prevalent microbe amplicons. Results: One hundred five genera and 117 species were detected in 50 ticks. Rickettsia , Anaplasma , Borrelia , Coxiella , and Ehrlichia were identified in tick samples by high-throughput sequencing at the genus level. Pathogen prevalence testing showed that the ticks were positive for B. burgdorferi s.l. (9/50 [18%]), A. phagocytophilum (1/50 [2%]), and the spotted fever group rickettsiae (35/50 [70%]), which were closely related to Candidatus R. jiangxinensis . Conclusion: The spotted fever group rickettsiae was the dominant pathogens in Xishuangbanna. Dual co-infection (1/50 [14%]) with Candidatus R. jiangxinensis and Borrelia garinii was demonstrated. V3-V4 high-throughput sequencing of the 16S rRNA gene was not sensitive to identify species for some bacteria, so more accurate and comprehensive analysis is necessary.

Ticks are one of the most important vectors that can transmit pathogens to animals and human beings. This study investigated the dominant tick-borne bacteria carried by ticks and tick-borne infections in forestry populations in Arxan, Inner Mongolia, China. Ticks were collected by flagging from May 2020 to May 2021, and blood samples were collected from individuals at high risk of acquiring tick-borne diseases from March 2022 to August 2023. The pooled DNA samples of ticks were analyzed to reveal the presence of tick-borne bacteria using high-throughput sequencing of the 16S rDNA V3–V4 region, and species-specific polymerase chain reaction (PCR) related to sequencing was performed to confirm the presence of pathogenic bacteria in individual ticks and human blood samples. All sera samples were examined for anti-SFGR using ELISA and anti-B. burgdorferi using IFA and WB. A total of 295 ticks (282 Ixodes persulcatus and 13 Dermacentor silvarum) and 245 human blood samples were collected. Rickettsia, Anaplasma, Borrelia miyamotoi, and Coxiella endosymbiont were identified in I. persulcatus by high-throughput sequencing, while Candidatus R. tarasevichiae (89.00%, 89/100), B. garinii (17.00%, 17/100), B. afzelii (7.00%, 7/100), and B. miyamotoi (7.00%, 7/100) were detected in I. persulcatus, as well the dual co-infection with Candidatus R. tarasevichiae and B. garinii were detected in 13.00% (13/100) of I. persulcatus. Of the 245 individuals, B. garinii (4.90%, 12/245), R. slovaca (0.82%, 2/245), and C. burnetii (0.41%, 1/245) were detected by PCR, and the sequences of the target genes of B. garinii detected in humans were identical to those detected in I. persulcatus. The seroprevalence of anti-SFGR and anti-B. burgdorferi was 5.71% and 13.47%, respectively. This study demonstrated that Candidatus R. tarasevichiae and B. garinii were the dominant tick-borne bacteria in I. persulcatus from Arxan, and that dual co-infection with Candidatus R. tarasevichiae and B. garinii was frequent. This is the first time that B. miyamotoi has been identified in ticks from Arxan and R. solvaca has been detected in humans from Inner Mongolia. More importantly, this study demonstrated the transmission of B. garinii from ticks to humans in Arxan, suggesting that long-term monitoring of tick-borne pathogens in ticks and humans is important for the prevention and control of tick-borne diseases.

目的 采用16S rDNA V3-V4区高通量测序,并结合特异基因的PCR检测,研究新疆维吾尔 自治区喀什地区亚洲璃眼蜱体内菌群特征及常见病原菌携带情况,为当地蜱传疾病防控提供理论依据.方法 于2021年5月在喀什地区巴楚县采用布旗法采集游离蜱342只,经形态学和特异性16S rDNA检测鉴定蜱种;除228只用于分离培养外,其余114只提取全基因组DNA,采用Illumina Novaseq测序平台进行16S rDNA V3-V4区高通量测序,分析喀什地区亚洲璃眼蜱携带菌的种类及其相对丰度水平;根据新疆地区既往报道常见蜱媒病原菌分布情况及蜱种带菌特征,选取6种常见病原菌对其特异性基因进行PCR检测,分析各病原菌的携带情况,并与高通量测序结果进行比较分析.结果 16SrDNA V3-V4区高通量测序分析显示1个样本携带贝纳柯克斯体;所有样本在属水平均检出立克次体属,16个样本检出无形体属,但缺少种水平注释;所有样本均未检出疏螺旋体.PCR法检测表明14只蜱携带伯氏疏螺旋体,7只蜱携带米氏疏螺旋体,3只蜱携带嗜吞噬细胞无形体,2只蜱携带贝纳柯克斯体,斑点热立克次体及查菲埃立克体未检出.结论 与特异基因的PCR法相比,16S rDNA V3-V4区高通量测序能够同时检测多种病原体,但也存在其检测受限于数据库、无法检测到种水平等缺点.因此,在病原监测的实际应用中,两种方法应相互结合,才能更全面、准确的反应当地蜱媒病原菌的分布情况.

Objective Ticks as ectoparasites might carry many pathogens and transmitted to their hosts through blood feeding. Human might have the opportunity to be bite by ticks, therefore getting various tick-borne pathogens. Distribution of bacterium flora in ticks plays an essential role in mapping and preventing local tick-borne diseases. Our aim is to map the distribution of bacterium flora and identify the main tick-borne pathogens in dominant tick species in grazing areas of Yunnan province. Methods V3-V4 region of 16S rRNA amplifier sequencing was used to analyze and study the tick-borne pathogens in Rh. Microplus. The prevalence survey on B.burgdorferi s.l., B. miyamotoi, E. chaffeensis, A. phagocytophilum, Coxiella burnetii and Spotted fever group rickettsiae was carried out via PCR. Results The results showed 105 genus and 117 species were detected in 50 (24 in Yunpan, 26 in Menghun) ticks. Pathogen prevalence testing showed the ticks were positive for B.burgdorferi s.l.(9/50, 18%), Spotted fever group rickettsiae (35/50, 70%), and A.phagocytophilum (1/50, 2%). Conclusion In our analysis, Spotted fever group rickettsiae were the dominant pathogens in these areas. At the same time, we found that v3-v4 high-throughput sequencing of 16S rRNA gene is not sensitive to identify species for certain bacteria(such as B. burgdorferi s.l.). More accurate and comprehensive analysis would be necessary for detecting tick bacterium flora.

Outbreaks of zoonotic diseases are accelerating at an unprecedented rate in the current era of globalization, with substantial impacts on the global economy, public health, and sustainability. Alien species invasions have been hypothesized to be important to zoonotic diseases by introducing both existing and novel pathogens to invaded ranges. However, few studies have evaluated the generality of alien species facilitating zoonoses across multiple host and parasite taxa worldwide. Here, we simultaneously quantify the role of 795 established alien hosts on the 10,473 zoonosis events across the globe since the 14th century. We observe an average of ~5.9 zoonoses per alien zoonotic host. After accounting for species-, disease-, and geographic-level sampling biases, spatial autocorrelation, and the lack of independence of zoonosis events, we find that the number of zoonosis events increase with the richness of alien zoonotic hosts, both across space and through time. We also detect positive associations between the number of zoonosis events per unit space and climate change, land-use change, biodiversity loss, human population density, and PubMed citations. These findings suggest that alien host introductions have likely contributed to zoonosis emergences throughout recent history and that minimizing future zoonotic host species introductions could have global health benefits.

目的 了解内蒙古阿尔山地区游离蜱的伯氏疏螺旋体感染状况及携带的基因型,为当地莱姆病的防控提供依据.方法 2020年5月和2021年5月,在内蒙古阿尔山地区用布旗法采集游离蜱,经形态学和特异性16SrDNA基因检测鉴定蜱种.随机选择113只蜱提取基因组DNA,采用巢式PCR扩增伯氏疏螺旋体5S～23S rRNA基因间隔区,阳性产物测序分析.采用BSK Ⅱ培养基对102只全沟硬蜱中携带的伯氏疏螺旋体进行分离培养,分离出的菌株采用多位点序列分析法(MLSA)鉴定基因型.结果 共采获游离蜱295只,经鉴定主要为全沟硬蜱(282/295,95.59％),其次为森林革蜱(13/295,4.41％).113只蜱的PCR检测结果显示,100只全沟硬蜱中24只携带伯氏疏螺旋体,感染率为24.00％,其中17只感染Borrelia garinii基因型(17/24,70.83％),7只感染B.afzelii基因型(7/24,29.17％),13只森林革蜱中未检出伯氏疏螺旋体.对102只全沟硬蜱中携带的伯氏疏螺旋体进行分离培养,得到2株伯氏疏螺旋体菌株,经MLSA分型鉴定为B.garinii基因型.结论 内蒙古阿尔山地区5月游离蜱以全沟硬蜱为主,其中伯氏疏螺旋体感染率为24.00％,感染的基因型为B.garinii和B.afzelii 2种致病基因型,并分离出2株B.garinii菌株,证实内蒙古阿尔山地区存在伯氏疏螺旋体的自然感染.当地应关注伯氏疏螺旋体的感染和致病风险,开展人群、媒介蜱和宿主的监测工作,建立科学合理的莱姆病防控体系.

Objective: To understand the infectious status of Lyme disease among patients with arthritis symptoms in Hainan Province, and to provide a theoretical basis for prevention and control of Lyme disease. Methods: From 2013 to 2018, sampling surveys had been conducted in medical institutions in 8 cities in Hainan Province(Haikou, Sanya, Danzhou, Dongfang, Wenchang, Qionghai, Qiongzhong, Wuzhishan), 2 311 patients serum samples were collected with arthritis symptoms, and descriptive research were conducted base on the collected clinical data. The Indirect Fluorescent-Antibody Test (IFA) method was used for preliminary screening of Lyme disease antibody, the Western Blot (WB) method was used for IFA positive samples confirmation. Statistical analysis using χ2 test. Results: 2 311 serum samples were tested by IFA, and 166 were positive with the positive rate of 7.18%. Further confirmed by WB method, 62 samples were positive, the positive rate of Lyme disease antibody was 2.68%(62/2 311). The positive rate of Lyme disease antibody among patients with arthritis in different regions of Hainan was statistically significant (χ²=40.636,P<0.001), and the positive rate in Qiongzhong city was the highest (8.81%, 14/159). Danzhou's positive rate was the second highest, 5.62%(5/89). Dongfang city had the lowest positive rate (0.51%, 2/394). The positive rates of Lyme disease serum antibody in men and women were 2.79% (33/1 182) and 2.57% (29/1 129), respectively; the positive rates of antibodies between each age groups were in the range of 1.74% to 3.64%. The antibody positive rate of Lyme disease showed no significant difference between gender and age (χ²=0.110,P=0.740 ;χ²=1.938,P=0.747). Conclusion: Patients with arthritis symptoms caused by Borrelia burgdorferi infection were found in 8 cities in Hainan province, but the Lyme disease antibody positive rate was different among cities, with Qiongzhong County being the highest.

目的 利用16S核糖体DNA(16S rDNA)全长测序方法,结合实验室检测数据,研究蜱中病原微生物种群特点,为病原体控制提供依据.方法 选取山西省祁县优势蜱种50只,经形态学和基因鉴定后,提取全基因组DNA,采用PacBio平台、高通量16S rDNA全长测序方法,分析蜱中微生物种类及各种群丰度情况;根据16S rDNA全长序列分析结果,利用PCR检测方法对可能存在的蜱媒病原体特异性基因进行鉴定,较为全面地分析蜱媒病原体携带情况.结果 16S rDNA全长测序分析表明50只血蜱均携带贝氏柯克斯体、假单胞菌,8只血蜱携带嗜吞噬细胞无形体,有部分血蜱携带立克次体、埃立克体,但仅匹配到属一级的分类信息,无法确定所携带的基因种,在50只血蜱中未检出疏螺旋体.PCR检测结果表明,50只血蜱中检测到柯克斯体内共生体、伯氏疏螺旋体、米氏疏螺旋体和斑点热群立克次体.结论 高通量16S rDNA全长测序分析可以同时检测出多种病原体,可提示当地的主要蜱媒致病菌,但也存在检测不全面、无法匹配到物种的缺点,与病原体特异检测方法相结合,能更全面地了解当地血蜱携带病原体的状况.

目的 建立双重荧光定量PCR方法,同时检测Borrelia burgdorferi和Borrelia miyamotoi.方法 分别利用recA基因、glpQ基因合成引物探针,并优化反应条件.在单重实验的基础上建立双重荧光定量PCR方法,对该方法灵敏度和特异度进行检测,并探索其他因素对反应结果的影响.使用模拟样本和实际样本评价新方法的检测效用.结果 双重荧光定量PCR方法具有良好的灵敏度和特异度,与单重实验方法比较,检测Ct值无明显差异(P＞0.05),病原浓度与检测Ct值具有良好的线性相关关系.两种病原浓度差异和其他病原的存在对检测结果无明显影响.使用该方法检测模拟蜱样本和实际蜱样本具有良好的实用性.结论 本实验建立的多重荧光定量PCR方法能快速便捷的检测两种疏螺旋体,为蜱样本病原鉴定提供了有价值的检测工具.

Lyme disease (LD) is one of the most important vector-borne diseases worldwide. However, there is limited information on the prevalence and risk analysis using correlated factors in the tropical areas. A total of 1583 serum samples, collected from five hospitals of Hainan Province, were tested by immunofluorescence assay (IFA) and western blot (WB) analyses using anti-Borrelia burgdorferi antibodies. Then, we mapped the distribution of positive rate (by IFA) and the spread of confirmed Lyme patients (by WB). Using ArcGIS, we compiled host-vector-human interactions and correlated data as risk factor layers to predict LD risk in Hainan Province. There are three LD hotspots, designated hotspot I, which is located in central Hainan, hotspot II, which contains Sanya district, and hotspot III, which lies in the Haikou-Qiongshan area. The positive rate (16.67% by IFA) of LD in Qiongzhong, located in hotspot I, was higher than that in four other areas. Of confirmed cases of LD, 80.77% of patients (42/52) whose results had been confirmed by WB were in hotspots I and III. Hotspot II, with unknowed prevalence of LD, need to be paid more attention considering human-vector interaction. Wuzhi and Limu mountains might be the most important areas for the prevalence of LD, as the severe host-vector and human-vector interactions lead to a potential origin site for LD. Qiongzhong is the riskiest area and is located to the east of Wuzhi Mountain. In the Sanya and Haikou-Qiongshan area, intervening in the human-vector interaction would help control the prevalence of LD.

目的 了解海南省西部地区关节炎或神经性疾病患者的莱姆病感染状况.方法 在东方市收集莱姆病疑似病例血清400份,儋州市收集莱姆病疑似病例血清500份,均为关节炎或神经性疾病患者.采用间接免疫荧光法(IFA)对血清标本进行莱姆病抗体初筛检测;并采用免疫印迹法(WB)对IFA检测阳性的血清标本进行确诊实验.结果 IFA方法检测血清样本900份,35份阳性,阳性率为3.89％.应用WB法对上述35份IFA阳性血清样本进行检测,结果15份阳性,阳性率为1.67％.不同年龄人群均有莱姆病螺旋体感染,其中,≤20岁阳性率最高,为11.76％(4/34),差异有统计学意义(x2=24.108,P＜0.001);莱姆病螺旋体感染在性别和症状间差异无统计学意义(x2=0.001,P=0.975;x2=0.011,P=0.917).结论 证实了海南省西部地区人群中存在莱姆病,建议当地医生在诊治关节炎和神经性疾病患者时,应排除莱姆病的可能.

莱姆病是一种重要的新发传染病,主要由蜱叮咬人、兽而传播.其病原体为伯氏疏螺旋体.人感染伯氏疏螺旋体的风险与蜱种的分布、地域因素、人为活动有关.该文对莱姆病的流行状况、基因型分布、主要传播媒介、动物宿主以及预防控制措施进行综述,为莱姆病的监测和防控提供参考.

目的 调查新疆维吾尔自治区(新疆)古尔图地区长尾黄鼠汉赛巴尔通体、伯氏疏螺旋体和嗜吞噬细胞无形体等病原体感染情况,分析该地区自然疫源性疾病流行风险,为制定防控措施提供科学依据.方法 2017年5-9月采用一日弓形夹法捕捉查岗果勒、布兰布拉克、白石头等地长尾黄鼠86只,无菌采集鼠体肾脏,试剂盒法提取全基因组DNA;运用实时荧光定量聚合酶链式反应方法检测样本汉赛巴尔通体ssrA基因、伯氏疏螺旋体recA基因和嗜吞噬细胞无形体Msp2基因.结果 共检测86只长尾黄鼠样本,其中汉赛巴尔通体阳性42份,阳性率为48.84％;伯氏疏螺旋体阳性6份,阳性率为6.98％;嗜吞噬细胞无形体阳性3份,阳性率为3.49％.布兰布拉克地区的汉赛巴尔通体感染率高于查岗果勒和白石头地区,白石头地区的伯氏疏螺旋体和嗜吞噬无形体感染率高于其他两地.结论 新疆古尔图地区长尾黄鼠存在汉赛巴尔通体、伯氏疏螺旋体和嗜吞噬细胞无形体感染.

Objective:To clone and express the 126-274 aa OspA peptide (OspA-pep) of Chinese Borrelia garinii ( B. garinii) strain PD91 and to preliminarily study its immune protectivity. Methods:The gene encoding the 126-274 aa OspA-pep of B. garinii PD91 was amplified by polymerase chain reaction (PCR) and then cloned into the prokaryotic expression vector pET-30a to construct the recombinant plasmid pET-30a-OspA-pep. Escherichia coli BL21 (DE3) competent cells transfected with the recombinant plasmid were induced by IPTG to express the target protein. The recombinant OspA-pep (rOspA-pep) was purified with Ni-IDA resin chromatography and its immunogenicity was analyzed by Western blot. New Zealand white rabbits were immunized with different doses of rOspA-pep (20, 30, 40, 50, 60, 80 and 100 μg). The titers of specific IgG antibodies in rabbit serum samples before and after immunization were detected by indirect immunofluorescence assay (IFA). The optimal immune dose was determined according to the antibody titer after immunization. In vitro neutralization test was performed to detect the immune protection of rOspA-pep using serum samples of the optimal immunization group. The optimal dose of rOspA-pep was used to immunize New Zealand white rabbits to observe the changes in antibody titer. Results:The recombinant plasmid pET-30a-OspA-pep was successfully constructed and highly expressed in host bacteria. Western blot showed that rOspA-pep had obvious antigen-antibody reaction with polyclonal antibody against B. garinii PD91 strain. IFA results showed the titers of IgG antibody in serum samples of rabbits immunized with rOspA-pep increased significantly (up to 1∶2 480) and 40 μg was the optimal dose. The neutralization rates of antibodies induced by 40 μg of rOspA-pep were 100% against 10 6 strain/ml of representative B. garinii PD91 and Borrelia afzelii ( B. afzelii) FP1 strains, 100% against 10 7 strain/ml of FP1 strain, and 60% against 10 7 strain/ml of PD91 strain. After immunization with 40 μg rOspA-pep on 1 d and 30 d, the titers of specific IgG antibody in rabbit serum samples reached the peak within two months, and maintained at that level for about 3-4 months before a gradual decline. Conclusions:The 126-274 aa OspA peptide fragment of Chinese B. garinii PD91 strain possessed good immunogenicity and induced antibodies with better in vitro neutralizing activity, which suggested that it could be used as a candidate component of the second generation subunit vaccine in China.

目的 了解海南省琼中地区媒介蜱伯氏疏螺旋体的感染状况,为当地莱姆病的防控提供科学依据.方法 在海南省琼中地区收集牛寄生蜱120只,应用普通PCR法扩增16S rRNA基因并测序分析,鉴定蜱种.应用巢式PCR方法,检测蜱的伯氏疏螺旋体带菌率,并对测序结果进行同源性比较以确定基因型.结果 本次海南省琼中地区牛寄生蜱经鉴定均为微小牛蜱.在120只蜱标本中,有44只蜱伯氏疏螺旋体检测阳性,阳性率为36.67％.同源性分析表明,琼中地区的微小牛蜱可感染多种基因型的伯氏疏螺旋体,包括Borrelia afzelii(56.82％)、B.yangze(25.00％)和B.garinii(18.18％)基因型.结论 琼中地区微小牛蜱中携带多种基因型的伯氏疏螺旋体,并且带菌率较高,有传播莱姆病的风险,应进一步加强当地人群和宿主的莱姆病调查和评估.

目的 了解海南省东北部地区人群莱姆病感染状况,为临床医生诊断莱姆病提供参考,为本省莱姆病的流行病学调查提供可靠的理论依据. 方法 收集海南省东北部地区三家医院1 334例有关节症状和(或)神经症状的患者血清,采用两步法检测莱姆病抗体,使用间接免疫荧光法(indirect fluorescent antibody test,IFA)初筛血清,对IFA检测阳性血清用免疫印迹法(Western blot,WB)确证. 结果 IFA法筛查海南省东北部地区1 334例患者莱姆病抗体阳性60例,阳性率4.50％.海口、文昌和琼海地区的三家医院该类就诊患者莱姆病抗体阳性率分别为4.25％、1.40％和8.28％ (IFA).三家医院阳性率比较差异有统计学意义(x2=25.640,P＜0.05).按不同性别、年龄分组海南省东北部地区三家医院莱姆病抗体阳性率比较差异均无统计学意义(x2=2.306,P＞0.05;x2=1.015,P＞0.05).WB法确证莱姆病抗体阳性28例,其抗体以31 KD的外膜蛋白A、33 KD-36 KD外膜蛋白B、39 KD膜脂蛋白、41 KD鞭毛蛋白和60 KD热休克蛋白抗体为主.结论 海南省东北部地区存在人群感染莱姆病,应加强当地疾病预防和监测.

目的 调查云南省剑川县野鼠中伯氏疏螺旋体(莱姆病螺旋体)的感染状况,以评估其对人群的潜在威胁.方法 2017年1-4月在剑川县选取石龙、长乐、新松3个自然村作为调查点,用鼠笼法进行捕鼠;经鼠种鉴定后采集鼠肾脏及膀胱标本,用BSK-Ⅱ培养基对其进行分离培养;用实时定量PCR及巢式PCR分别检测伯氏疏螺旋体特异性recA基因及5S～23S rRNA基因间隔区.结果 ①共捕获10种439只鼠,其中齐氏姬鼠219只、大绒鼠165只,其余鼠种55只(中华姬鼠24只、贝氏树鼩13只、斯氏家鼠7只、巢鼠3只、灰麝鼩3只、赤腹松鼠2只、社鼠2只及大足鼠1只);②439份标本均未分离培养到伯氏疏螺旋体;③实时定量PCR检测,结果显示伯氏疏螺旋体总阳性率为2.51％(11/439),其中中华姬鼠4.17％(1/24)、齐氏姬鼠4.11％(9/219)和大绒鼠0.61％(1/165);④11份阳性标本皆来源于剑川县石龙村,其余2个村的鼠标本未检测到阳性;⑤11份实时定量PCR阳性标本的5S～23S rRNA基因间隔区的检测皆为阴性.结论 云南省剑川县野鼠中存在莱姆病螺旋体的自然感染,但鼠中病原体的载量较低,其传播给人群的风险不大.