海南省399例斑点热群立克次体的四种实验方法检测结果分析  

Analysis on the results of 399 cases about SFGR's four expertmental methods in Hainan Province

作　　者：吴捷[1] 王少玲[1] 马焱[1] 曾祥洁[1] 邱丽[1] 贾鹏本[1] 华德[1] 李沅[1] 聂绍发[2] 

机构地区：[1]海南省疾病预防控制中心微生物检验科,海南海口570203 [2]华中科技大学同济医学院公共卫生学院流行病学与卫生统计学系,湖北武汉430030

出　　处：《中华疾病控制杂志》2016年第2期184-187,共4页Chinese Journal of Disease Control & Prevention

基　　金：海南省自然科学基金(808225)

摘　　要：目的了解海南省不明原因发热病人中斑点热群立克次体的感染情况和流行病学特点,分析立克次体病实验检测方法的特点,拟建立一种快速、特异性强、灵敏度高、重复性好的实验室检测技术,为斑点热的防控提供科学依据。方法分别采用巢式聚合酶链式反应(polymerase chain reaction,PCR)、多重实时荧光定量PCR、间接免疫荧光法和病原学分离培养法对不明原因发热病人血标本进行检测。结果用巢式PCR检测发热病人399份血标本中的gro EL基因的特异性片段,检测出17份斑点热群及斑疹伤寒群立克次体阳性,61份恙虫病东方体阳性。用多重实时荧光定量PCR检测出2份斑点热群立克次体阳性,6份斑疹伤寒群立克次体阳性,49份恙虫病东方体阳性。用间接免疫荧光实验检测,斑点热群立克次体Ig M阳性结果为19份,Ig G为148份;斑疹伤寒群立克次体Ig M阳性结果为29份,Ig G为169份。采用L929单层细胞对血标本进行病原学分离培养,培养结果为阴性。结论四种实验室检测方法从病原学、血清学和分子生物学方面进一步阐明斑点热的自然疫源性特点,为其诊断及防护提供有力的实验室依据。Objective To investigate the infection status and epidemiological features of spotted fever group Rickettsiae（ SFGR） in patients with fever of unknown origin（ FUO） in Hainan Province,and analyze the characteristics of detection of rickettsial disease,so as to establish a rapid strong specificity,high sensitivity and good repeatability laboratory testing system which would provide scientific basis for prevention and control of the spotted fever.Methods Nested polymerase chain reaction（ Nested PCR,） multiple real-time fluorescence quantitative PCR,indirect immunofluorescence and pathogen isolation culture methods were conducted respectively to inspect the blood samples of patients with FUO.Results Nested PCR was performed to detect the specific fragment of gro EL gene in 399 blood specimens of fever patients.The results showed that 17 samples contained spotted fever group and typhus group rickettsiae,61 samples contained orientia tsutsugamushi.Meanwhile,the result of multiplex real-time fluorescence quantitative PCR test,showed that 2 samples contained spotted fever group rickettsiae,6 samples contained typhus fever group rickettsiae,49 samples contained orientia tsutsugamushi.Furthermore,when indirect immunofluorescence assay were performed,SFGR Ig M were observed in 19 samples and Ig G were observed in 148 samples.Typhus fever group rickettsiae Ig M were observed in 29 samples,and Ig G in169 samples.However,no cytopathic effect and negative results were showed in the test isolated and cultured the blood specimens with L929 monolayer cells.Conclusions Four kinds of laboratory testing methods could further clarify the epidemic characteristics of spotted fever from aspects of etiology,serology and molecular biology,provide plenty data for diagnosis and protection.

关 键 词：立克次体科感染 实验室技术和方法 流行病学研究 

分 类 号：R183.5[医药卫生—流行病学] R181.1[医药卫生—公共卫生与预防医学]