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目的 评价基因扩增法应用于检测鼠类标本调查恙虫病疫源地的作用。方法 用一定量恙虫病东方体人工感染昆明种小鼠 ,攻击后第 3、6、9天处死小鼠取血块及脾脏用特异性套式 PCR技术进行恙虫病东方体 DNA的检测 ,再用该技术测定现场采集的鼠类标本 ,以可从标本中扩增出一段长 88bp的 DNA片段作为实验或现场捕获鼠感染了恙虫病东方体的指标 ,继而估计该地区是否存在恙虫病疫源地。结果 实验感染恙虫病东方体 3天时在两类标本中均不能检出该病原 DNA,而第6天测定时均可发现含有恙虫病东方体 ;从福建闽西北地区采集的 111份鼠类脾脏中检出 1份阳性标本 ,另从江西送检的 2 9份野鼠血块中也检出 1份阳性标本 ,说明采集标本区域已成为恙虫病疫源地。结论 套式 PCR扩增技术具有很好的特异性和敏感性 ,可用于现场鼠类标本的恙虫病病原学的调查。
Objective To evaluate the role of gene amplification in the detection of tsutsugamushi disease in mice. Methods Kunming mice were artificially infected with a certain amount of Orientia tsutsugamushi and the mice were sacrificed on the 3rd, 6th and 9th days after the challenge. Blood samples were taken from the blood clots and spleens of the tsutsugamushi. The technique was used to determine the presence of tsutsugamushi disease in the area by detecting a murine specimen collected on site to amplify a DNA fragment of 88 bp in length from the specimen as an experimental or on-site capture rat infected with Orientia tsutsugamushi. Results Experimental tsutsugamushi infected Oriental body 3 days in both types of specimens were not detected in the pathogenic DNA, and the determination of the first 6 days can be found in tsutsugamushi Oriental body; collected from Fujian, Fujian Province, 111 northern mouse spleen One positive specimen was detected. Another positive specimen was also detected in 29 wild rat blood clots submitted by Jiangxi Province, indicating that the specimen collection area has become the source of scrub typhus. Conclusion The nested PCR amplification technique has good specificity and sensitivity and can be used to investigate the etiology of scrub typhus in the live rodents.