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目的对海南省澄迈县进行斑点热自然疫源地现场调查。方法以病原学证实的首发患儿居住村外延收集可疑病例并采集当地野生啮齿动物抗凝血,用现代Shellvial培养技术进行立克次体病原分离,对分离株进行16S rRNA、gltA、ompA、17kD蛋白基因及groEL基因扩增并测序分析。以WHO立克次体协作中心推荐间接免疫荧光试验检测患者、全县10村812名农业人群、18只家犬及44只野生啮齿动物血清斑点热群立克次体抗体。巢式PCR扩增患者、家犬、野生啮齿动物、媒介蜱、螨及蚤标本groEL基因并测序分析。分析斑点热群立克次体优势流行株及可能的媒介及宿主种类以及人群及家畜对当地斑点热群立克次体优势流行株抗体阳性率。结果从调查的21例可疑立克次体患者血液标本中分离到11株斑点热群立克次体,从15只野生针毛鼠血液标本中分离到7株斑点热群立克次体,遗传学分析证实分离株与黑龙江立克次体及广东野鼠斑点热群立克次体分离株关系密切,但血清表型不同黑龙江立克次体。犬血、媒介蜱、螨及蚤PCR扩增groEL基因及测序结果支持病原学证据并且序列分析结果显示与病原分离株为同一流行株。血清学结果提示当地人群对该流行株IgG抗体总阳性率为46.1%,家犬88.9%,鼠11.4%。发热可疑患者强力霉素经验治疗痊愈。结论正常人群、发热患者、媒介及宿主等病原学、分子生物学及血清学调查结果证实海南省存在新发斑点热疫源地,加强临床诊断及鉴别诊断,进一步监测其他蜱传立克次体病,消杀农户家居及家畜体表媒介蜱,防止叮咬是当地防治该病的关键。
Objective To investigate the spot hot natural foci in Chengmai County, Hainan Province. Methods Susceptible cases were collected from the outpatients in the first trimester of children with pathologically confirmed anaphylaxis and local anticoagulant rodents were collected. The Rickettsial pathogens were isolated using modern Shellvial culture technique. The isolates were analyzed for 16S rRNA, gltA, ompA, 17kD Protein gene and groEL gene amplification and sequencing analysis. WHO Rickettsia Cooperative Center recommended indirect immunofluorescence test for detection of patients, the county 812 agricultural population, 18 dogs and 44 wild rodent serogroup hot spot rickettsia antibody. The groEL gene was amplified by nested PCR in dogs, domestic dogs, wild rodents, vector ticks, mites and fleas and sequenced. The prevalence of dominant epidemic strains of Rickettsia and the possible media and host species of the speckle hot group as well as the prevalence of antibody positive to dominant epidemic strains of Rickettsia from the local spotted fever group were analyzed. Results Eleven Spotted Rickettsia species were isolated from the blood samples of 21 suspected Rickettsia patients. Seven Rickettsia species were isolated from the blood samples of 15 wild Needle Mice, The analysis confirmed that the isolates were closely related to Rickettsia and Rattus norvegicus in Guangdong Province, but the serological phenotype was different from that of Rickettsia in Heilongjiang Province. PCR amplification of groEL gene and sequencing results in canine blood, ticks, mites and fleas supported etiological evidence and sequence analysis revealed the same strain as the pathogenic isolate. Serological results suggest that the local population of the prevalence of IgG antibodies in the prevalence of 46.1%, 88.9% dogs, rats 11.4%. Fever patients suspected treatment of doxycycline cured. Conclusion The etiological, molecular and serological findings of normal population, fever patients, vector and host confirmed the existence of new plague origin in Hainan Province, strengthened clinical diagnosis and differential diagnosis, and further monitored other tick-transmitted rickettsia Diseases and killings Households and livestock body surface ticks ticks, prevent bites is the key to local prevention and treatment of the disease.