【作者】 支慧；

【导师】 王克霞；

【作者基本信息】 安徽理工大学 ， 病原生物学， 2006， 硕士

【摘要】 安徽省因其独特的地理、气候环境及农业习俗的沿用至今,使其成为我国钩端螺旋体病(Leptospirosis,以下简称为钩体病)的重灾区之一。而钩端螺旋体(Leptospira,以下简称为钩体)的宿主多样化、血清群/型多样性也增加了对其进行预防控制的难度。因此,对钩体菌群在安徽省的分布情况进行调查,以分析及预测安徽省的钩体病的流行状况具有重要的理论意义;同时我们将对其新近于安徽省发现的新血型钩体进行实验研究,以明确其在流行病学中的意义。 在本次研究中,我们将2000年到2005年从各下级市县疾病预防控中心送检的自然人群血清标本与我国现流行的15群15型做MAT(显微镜凝集实验),对其进行分型鉴定。我们发现安徽省的钩体病的病发得到了比较好的控制,标本的平均阳性率为6.14%,不过我们亦发现其有轻微的上升。与此同时我们认为从90年代起就出现了钩端螺旋体菌群更迭的现象—南黄北移,在这五年中有了进一步的发展,并已经造成了安徽省北部地区的菌群构成比例的改变,而安徽省南部地区的钩体菌群的多样性更为明显,原先占于首位的黄疸出血群已经有被替代的趋势。同时,我们将传统的血清学手段、分子生物学方法及病理学观察相结合,对2003年于安徽省皖南山区会线蛙肾中分离培养的一新群新型(顾群秦型)钩端螺旋体的代表株EG79对行了综合性分析研究。其中传统的血清学方法进行分析结果显示:2,6-DAP不能抑制EG79的生长,EG79株能在13℃和8-AG中生长,EG79株不能在37℃生长,对铜离子敏感,与1mol/L NaCl有反应。从PFGE图谱上看出EG79与腐生菌的图谱相对于与致病性菌株的图谱有明显的不同,在对注射了EG79的豚鼠进行观察,体重、体温改变有统计学意义;PCR检测结果显示实验组豚鼠的肝、肾中有钩体存在,且显示出致病性钩体特异性的285bp这一片段。解剖后我们发现其肺部有明显的出血现象,肝脏、肾脏均有肿大,且镜下观察发现肺泡、肾小球、心肌都有红细胞的浸入;镜下发现肝脏有炎症细胞的浸润。 本文对安徽省近六年来钩体菌群分布的特点进行了持续性地调查,对其结果进行了分析,并对出现改变的可能性原因进行了阐述。同时,因为某一型钩体是属于致病性还是腐生性在对人畜的危害是不同的,因此对新型钩体进行研究并对其归属做出正确的结论对于发现这一新菌型的地区的钩体病的防治有着重要意义,本文多角度地对其进行了综合性的研究,在对研究结果进行分析后倾向于将这一新型钩体归于致病性钩端螺旋体中。更多还原

【Abstract】 AnHui Province has been estimated as a distress area of Leptospirosis because of its special geographic feature, climatop. And it is difficult to prevent and control leptospirosis because of so many kinds of hosts and serotypes. This time, we want to investigate the distribution of Leptospira that area from 2000 to 2005 to analysis and predict the epidemic situation of Laptospirosis in AnHui Province. So, we can offer the theoretical support to prevent and control the Leptospirosis in AnHui. At the same time, we will study the new serotype found in Wannan Mountain Area to evaluate the possibility of epidemity.In this thesis, we take the MAT to type the Leptospira isolated from the blood samples collected from 2000 to 2005, and then analysis the development tendency of the Leptosprosis in AnHui. After typing the Leptospira isolated from the collected blood samples from the inferior centers of disease control and prevention, we find that the Leptospirosis in AnHui has been controlled and the positive rate is 6.14%. But the results also show the slight upgrade of Leptosprosis and the icterohaemorrhagiae which has been found in the south of the AnHui province now has been found in the north.At the sime time we take a series of methods including the serology methods, biological genetic methods and pathology observation to study the representive strain of Gu serogroup, Qin serovar EG79, isolated form Ronan plancyi. The results of the orthodox serological methods to EG79 show: 2,6-DAP can not restrain the growth of EG79, EG79 can grow in 13℃ and the medium adding 8-AG while it can not grow in 37℃, EG79 is sensitive to Gu²⁺ and can react with 1mol/L NaCl. From the patterns of PFGE, we can find the obvious difference of the EG79 and the nonpathogenic Leptospira. The result of injecting the well-grown culture of the EG79 strain into guinea pigs showed that the viabilities of the infected guinea pig declined with rise of body temperature and reduction of body weight. Definite pathological lesions could be demonstrated in the liver and lungs. The PCR showed a positive result It is concluded that this new type of Leptospira interrogens possesses a certain degree of virulence to guinea pigs, but further experimentations are necessary to prove its virulence to human beings.Compare the information of Leptospira epidemic situation in 90s at AnHui Province, we thought that the distribution of Leptospira in AnHui has changed and the icterohaemorrhagiaewhich has been found in the south of the AnHui now has been found in the north. So, it is important to monitor persistently. And it is significant to identify whether a kind of Leptospira belongs to Leptospira intemogans, because only Leptospira interrogans can cause the Leptospirosis. From all of the results of the study to EG79, we prefer to identify the Gu serogroup, Qin serovar Gu serogroup, Qin serovar is a new kind of Leptospira interrogans. So we thought this study has theoretical sense, especially when we found the types of Leptospira carried by Ronan plancyi accord with that carried by local human.更多还原