六盘山地区蜱中莱姆病螺旋体检测及分型

目的 了解宁夏部分地区蜱中莱姆病螺旋体感染及基因型别情况。方法 选择宁夏六盘山地区为调查点,采集当地蜱类,用巢式PCR法进行检测,阳性产物进行限制性片段长度多态性(RFLP)分析,确定莱姆病螺旋体的基因型。结果 共检测4个蜱种206只蜱,阳性37只,阳性率17.96%;其中以青海血蜱阳性率最高,达59.37%;RFLP分析表明,蜱中莱姆病螺旋体均为B.garinii基因型,但型内存在一定差别,其中有29份标本属B.garinii基因型Ⅰ群,有8份标本属B.garinii基因型Ⅱ群。结论 证实在宁夏六盘山地区蜱中存在莱姆病螺旋体,并发现其属于B.garinii基因型。     

新疆北疆部分地区硬蜱伯氏疏螺旋体基因型研究

目的调查和鉴定新疆北疆石河子市、沙湾县、伊宁县和察布查尔县媒介蜱,并确定其携带的莱姆病螺旋体的基因型。方法选择4县市6个牧区为调查点,采集羊源寄生硬蜱,结合形态学与16S rRNA进行蜱种鉴定;采用BSK-H培养基对莱姆病螺旋体分离培养,并用硝酸银染色和巢式PCR法进行病原检测,阳性产物测序结果比对,并与11种GenBank参考序列比对,确定莱姆病螺旋体的基因型。结果6个调查点共采集寄生硬蜱900多只,经形态学与16S rRNA鉴定分别为图兰扇头蜱、刻点血蜱、亚洲璃眼蜱和边缘革蜱;从中分别选取样本硬蜱共102只,分24管,分离培养后结合巢式PCR和硝酸银染色共获得16管阳性培养物。5S～23S rRNA基因问隔区测序比对分析表明,所有菌株为伯氏疏螺旋体与国际报道的B.burgdoq''eri Sensu Stricto(B31)基因型具有高度同源性,同源性为98.6%～99.5%;OspC基因型分析与上述结果一致。结论4县市分离获得伯氏疏螺旋体,基因型为B.burgdooCeri Sensu Stricto,且存在生物多样性。从图兰扇头蜱中分离到伯氏疏螺旋体为我国首次报道。     

北京西山地区长角血蜱及其体内伯氏疏螺旋体实验观察

目的 :实验观察长角血蜱及其体内伯氏疏螺旋体形态结构。方法 :1994～ 1996年在北京西山地区采集长角血蜱 180只。标本应用蜱整体封片法、冰冻切片和石蜡切片结合HE染色和硝酸银染色法分别检测和光镜观察。结果 :蜱整体封片法直接镜检即可观察到蜱外部基本形态结构。HE染色观察蜱内部器官细微形态结构。冰冻切片和石蜡切片结合硝酸银染色 ,检出北京西山地区长角血蜱 (包括幼蜱、若蜱、成蜱和饱血蜱 )伯氏疏螺旋体感染率为 2 3 % ,显微镜下可清晰观察到蜱体伯氏疏螺旋体多种不同的形态。结论 :上述方法对莱姆病媒介蜱及伯氏疏螺旋体的流行病学研究有一定价值     

新疆维吾尔自治区蜱携带伯氏疏螺旋体流行特征分析

目的:分析新疆维吾尔自治区(简称新疆)蜱携带伯氏疏螺旋体的流行状况和分子特征。方法:2020年4 - 6月,在新疆伊犁、阿拉山口、呼图壁、青河、福海、五家渠6个地区采集312份硬蜱样本,应用巢式PCR法和荧光定量PCR法检测蜱携带伯氏疏螺旋体情况,对两种方法检测均为阳性的样本进行巢式PCR产物基因分型鉴定。结果:巢式P...     

贵州省四城市乙型肝炎病毒感染者病毒基因型调查研究

目的 调查贵州省4城市乙型肝炎病毒（HBV）感染者的病毒基因型及其与临床的关系。方法 选择贵阳、遵义、凯里、都匀4城市慢性HBV感染患者共786例，其中无症状携带者（ASC）346例，慢性肝炎（CH）313例，肝硬化（LC）77例，肝细胞肝癌（HCC）50例。用S基因限制性片段长度多态性确定基因型，直接测序分析B基因亚型，比较主要基因型地区分布及临床特征。结果 786例中，B基因型497例（63．23％），C型275例（34．99％），A型7例（0．89％），D型7例（0．89％），未发现E、F型。B型的分布：凯里市最高（96．04％），遵义、都匀市其次（78．79％、64．52％），贵阳市最低（53．14％）。C型的分布，贵阳（45．84％）高于都匀（34．41％）、遵义（13．13％）及凯里市（3．96％），差异有统计学意义（P〈0．05或P〈0．01）。94例B型感染者中，93例为Ba（98．94％）、1例为目亚型。从ASC、CH、LC到HCC组，B型的分布逐渐降低，而C型在各组的分布逐渐增高。与B型相比，C型感染者年龄较大；ALT水平较高（P〈0．05）；HBeAg阳性较低（P〈0．025）。结论 贵州省存在A、B、C、D4种HBV基因型，但以B型为主，C型其次，A、D型极少。B型中又以Ba亚型为主。B、C基因型在贵州省4城市的分布有一定差异。     

广西凭祥地区莱姆病螺旋体检测和基因分型研究

目的 调查广西凭祥地区莱姆病感染情况和基因型别.方法 2011年7月从广西凭祥地区采集蜱、啮齿动物和野鸟标本,分别采取煮沸法和Qiagen试剂盒提取蜱、啮齿动物脾脏以及鸟脾脏中莱姆病螺旋体基因组DNA;采用巢式PCR扩增莱姆病螺旋体5S～23S rRNA基因间隔区;对PCR扩增产物进行测序,并将序列结果与GenBank中莱姆病螺旋体5S～23SrRNA基因问隔区序列比对分析.结果 从3份啮齿动物标本中检测到莱姆病螺旋体5S～23SrRNA基因间隔区片段,啮齿动物的感染率为5.66％(3/53).其中一个序列与GenBank中Borrelia valaisiana基因型莱姆病螺旋体(序列号:HM100125.1,AB091455.1,AB091454.1,AB091453.1)同源性为100％;蜱和鸟标本中未检测到莱姆病螺旋体.结论 广西凭祥地区啮齿动物中存在莱姆病螺旋体B.valaisiana基因型感染.     

伯氏疏螺旋体传播过程中病原体、媒介、宿主间的相互作用

莱姆病是一种蜱媒传播的人兽共患病，在北半球广泛流行。其病原体是伯氏疏螺旋体，具有极为复杂的传播机制和过程。在过去20多年中，‘科学家们对此进行了广泛而深入的探索。本综述拟对近5年这方面的研究进展，尤其是Fikrig实验室的工作进行总结。内容主要包括：证明伯氏疏螺旋体外膜蛋白OspA是一种粘附素（adhesion），对螺旋体在硬蜱中肠（midgut）定居至关重要，并发现硬蜱中肠肠腔表面蛋白TROSPA是OspA的受体；发现蜱唾液腺蛋白Salp15协助伯氏疏螺旋体感染哺乳类动物；发现伯氏疏螺旋体膜蛋白bmpA和bmpB介导螺旋体在关节的定居及关节炎的发生。这些发现有助于我们深入了解莱姆病的传播特征和致病机制。     

微波免疫组化SP法在长角血蜱体内伯氏疏螺旋体形态研究中的应用

目的：观察长角血蜱体内伯氏疏螺旋体的免疫形态特征。方法：1996年5月，对北京西山采集的羊体表长角血蜱寄生蜱标本行微波免疫组化SP法检测和伯氏疏螺旋体形态观察。结果光镜下，寄生蜱中肠内显示伯氏疏螺旋体菌体数量多且形态多样。螺旋体螺旋波纹呈不规则锯齿形，或呈纤细疏螺旋形。短形螺旋体有3—10个疏螺旋，而长形螺旋体达30个以上的疏螺旋。结论：微波抗原修复免疫组化SP法染色，蜱中肠标本中伯氏疏螺旋体显示强阳性，易于伯氏疏螺旋体的检出和形态观察。     

用PCR/RFLP技术对黑龙江沿岸蜱类和鼠类中北亚蜱传斑点热的检测

为了解黑龙江沿岸部分地区斑点热自然疫源地存在情况，作者用PCR／RFLP的方法检测该地区蜱类及鼠类中的斑点热群（spottedfevergroup，SFG）立克次体。结果从该地区的森林革蜱、嗜群血蜱、黑线姬鼠、东方田鼠、麝鼠及棕背中均扩增出了斑点热群立克次体的特异片段，而对斑疹伤寒立克次体、恙虫病立克欢体、Q热立克次体则为阴性。PCR产物经PstⅠ及RsaⅠ酶切后发现它们的酶切图谱与西伯利亚立克次体完全相同，而有别于其它斑点热群立克次体标准株。作者认为黑龙江沿岸调查点可能存在北亚蜱传斑点热的自然疫源地。     

Borrelia burgdorferi Sensu Stricto DNA in Field-Collected Haemaphysalis longicornis Ticks,Pennsylvania, United States

We collected questing Haemaphysalis longicornis ticks from southeastern counties of Pennsylvania, USA. Of 263 ticks tested by PCR for pathogens, 1 adult female was positive for Borrelia burgdorferi sensu stricto, yielding a 0.4% infection rate. Continued monitoring of this invasive tick is essential to determine its public health role.     

Detection and typing of human papillomavirus DNA in paired urine and cervical scrapes

The prevalence of human papillomavirus (HPV) in paired cervical scrape and urine specimens from 144 women attending a clinic for genitourinary medicine was determined by polymerase chain reaction (PCR) and nested PCR, using degenerate and general primer pairs localized within the L1 region. HPV typing was by restriction fragment length polymorphism (RFLP), type-specific PCR (HPV 6, 11, 16, 18, 33), and partial DNA sequencing of PCR products. HPV DNA was detected in 114 (84%) women. HPV DNA was detected in the specimens of 58 patients after amplification with MY09/MY11 primers and in a further 54 patients after nested PCR with the GP5 + /GP6 + primers. A total of 106/136 (78%) of women had HPV DNA positive cervical scrapes and 89 (65%) had HPV DNA positive urine specimens. Both the urine and cervical specimens of 81 women were positive. In 25 women HPV DNA was detected in the cervical specimen only, and in 8 women HPV DNA was detected in the urine specimens only. A total of 108 specimens from 75 patients were typed. For 33 patients HPV typing was achieved in both the cervical and the urine specimens and 19 women had identical types in paired specimens. Multiple HPV infections could be detected in 15 (20%) of 75 women where either the cervical and urine specimen or both of the specimens could be typed. More then one HPV type was found in 8 specimens and from multiple sites (cervix and urinary tract) in the same patients on 7 occasions. The results of this study indicate that the detection of HPVs in the urogenital tract can be maximised through the testing of both cervical scrapes and urine specimens in conjunction with the use of a nested PCR to increase the sensitivity of HPV DNA detection. Also, urine cannot be a direct substitute for a cervical scrape as different HPV types are often detected in the urine compared with those detected in the cervix.     

Borrelia,Ehrlichia, and Rickettsia spp. in Ticks Removed from Persons,Texas, USA

Data regarding the type, frequency, and distribution of tick-borne pathogens and bacterial agents are not widely available for many tick species that parasitize persons in the southern United States. We therefore analyzed the frequency and identity of pathogens and bacterial agents in ticks removed from humans and subsequently submitted to the Texas Department of State Health Services, Zoonosis Control Program, from October 1, 2004, through September 30, 2008. The data showed associations of bacterial agents and potential vectors. Tick-related illnesses may pose unidentified health risks in areas such as Texas, where incidence of human disease related to tick bites is low but well above zero and where ticks are not routinely suspected as the cause of disease. Cause, treatment, and prevention strategies can be better addressed through collecting sufficient data to establish baseline assessments of risk.     

安徽省丙型肝炎病毒基因型分布

应用逆转录－聚合酶链反应和限制性片段长度多态性技术分析安徽省丙肝患者丙型肝炎病毒基因型。结果表明ＨＣＶ基因型有Ⅰ、Ⅱ、Ⅲ、Ⅳ及Ⅱ／Ⅲ混合型，以Ⅱ型感染（７５．６６％）优势，其次为Ⅲ型（１３．２３％）和Ⅱ／Ⅲ型（６．３５％），Ⅰ、Ⅳ型各有１例（０．５３％），安徽省北部地区的ＨＣＶ型多于南部，而Ⅱ型北部则少于南部，不同地理区域ＨＣＶ基因型分布差异有显著性（Ｐ〈０．０５），有、无输血、血液制品史的丙肝患     

Autochthonous Lyme Borreliosis in Humans and Ticks in Korea

ObjectiveThis study aimed at finding epidemiological and clinical features of autochthonous Lyme borreliosis in humans through epidemiological investigations and identifying its vectors and pathogens through analysis of ticks.MethodEpidemiological investigations, including review of the retrospective medical records and patient interviews, were conducted in two cases that occurred in 2012. To identify the vectors and pathogens, ticks were collected between September 23 and October 6, 2012 from the area where the tick bite in the first patient occurred. The ticks were classified, and polymerase chain reaction (PCR) tests and cultures were performed.ResultsThe first patient, a 46-year-old female, visited a forest in Gangwon province, which was 900 m above sea level, where the tick bite occurred. Two weeks after the tick bite, erythema migrans (12 × 6 cm² in size) appeared on the site of tick bite, along with fever, chill, fatigue, myalgia, and arthralgia on shoulders, knees, and hips. The second patient, a 44-year-old male, visited a mountain in Gangwon province, which was 1200 m above sea level, where a tick bite occurred. One month after the tick bite, erythema migrans appeared at the site of the tick bite, along with fatigue, myalgia, and arthralgia on the right shoulder and temporomandibular joint. Indirect fluorescent antibody testing and Western blotting were carried out in these two cases for diagnosis, and positive findings were obtained. As a result, Lyme borreliosis could be confirmed. To estimate the pathogens and vectors, the ticks were collected. A total of 122 ticks were collected and only two species, Haemaphysalis japonica and Haemaphysalis flava, were identified. PCR and culture were performed on ticks. However, Borrelia burgdorferi sensu lato was not isolated from any collected ticks.ConclusionsThis study is significant to confirm Lyme borreliosis officially at first by the national surveillance system, although identification of the mites and pathogens failed.     

霍乱弧菌4种新类型tcpA基因发现及序列分析

目的研究中国霍乱弧菌（VC）毒力协同调节菌毛A亚单位基因（tcpA）的多态性.方法分别进行聚合酶链反应及限制性片段长度多态性分析、型特异引物PCR分型、基因测序及序列分析.结果 200株O1群埃尔托型（EVC）产毒株和100株O139群VC产毒株的tcpA基因均与EVC国际标准株N16961株为同一类型（t2型）.50株EVC非产毒株中只有3株携带tcpA基因,1株为t2型,另2株为2种新类型.20株O139群VC非产毒株均不携带tcpA基因.20株非O1非O139群VC中只有2株携带tcpA基因,且为2种新类型.4种新类型tcpA基因与国外发现的4种类型比较,基因序列及推测的氨基酸序列同源性分别为58.3%～77.5%和61.0%～85.5%;5′端约102bp基因序列基本一致,推测的氨基酸序列完全一致;3′端约210bp基因序列变异最大,推测的氨基酸序列变异也最大.抗原表位分析,C末端差异最大.结论建立快速准确区分不同类型tcpA基因的方法,并发现4种新类型.     

广东省潮州市13750例妇女宫颈人乳头瘤病毒感染状况及基因型分析

目的对广东省潮州市13750例妇女进行人乳头瘤病毒(HPV)筛查,探讨该地区宫颈HPV感染现状及基因亚型分布情况,为该地区预防HPV感染和防治宫颈癌提供科学依据。方法 2010年3月至7月在广东省潮州市采取整群抽样法,选择潮州市市区及其2个乡镇中符合纳入标准并完成研究的13750例受试者为研究对象。采用导流杂交基因芯片技术对HPV基因亚型进行分型检测(本研究遵循的程序符合本院人体试验委员会所制定的伦理学标准,得到该委员会批准,征得受试对象本人的知情同意,并与之签署临床研究知情同意书)。结果自13750例受试对象中,共计检出989例HPV呈阳性患者,感染率为7.2%。高危型HPV感染的前5位HPV基因亚型依次为HPV-52,-16,-58,-68及-33,感染率分别为27.50%,15.47%,10.82%,8.29%和6.77%。其中,单一、合并2种、3种、4种、5种及以上HPV亚型感染分别为752例(76.1%),156例(15.8%),53例(5.4%),18例(1.9%)及9例(0.9%)。低文化程度,低收入家庭、从事家务劳动及嗜烟、酒受试者的HPV感染率较高。结论 HPV基因亚型分布具有一定区域性,高危型HPV-52,-16和-58及合并1种HPV基因亚型感染在广东省潮州市常见。