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目的探讨环介导同温DNA扩增法(LAMP)与解剖-显微镜检法对血吸虫感染不同阶段钉螺检出率的差异。方法用日本血吸虫毛蚴感染钉螺,于25℃培养箱中培养。分别于感染后1、2、3、4、5、6、7、8、9、10周收集10只实验钉螺,解剖后在解剖镜下检查血吸虫感染情况。取解剖的钉螺软体抽提基因组DNA,采用LAMP法进行检测。在感染后10周,取100只实验钉螺,同时用解剖-显微镜检法和LAMP法进行检测。收集吸虫病流行区现场钉螺,采用两种方法同时进行检测,统计血吸虫感染性钉螺阳性率。结果解剖-显微镜检法和LAMP法检测感染1~10周的10批钉螺血吸虫感染率分别为0.00%、0.00%、0.00%、10.00%、0.00、10.00%、20.00%、20.00%、60.00%、60.00%和50.00%、20.00%、30.00%、20.00%、20.00%、40.00%、30.00%、40.00%、80.00%、70.00%,总阳性率分别为18.00%和40.00%,差异有统计学意义(χ2=11.75,P<0.01)。两种方法检测感染后7周的100只钉螺阳性率分别31.00%和39.00%,检测来源于两个不同省份血吸虫病流行区钉螺,阳性率分别为29.80%和19.47%。结论环介导同温DNA扩增法能有效地检出感染不同阶段的感染性钉螺,检测结果能更准地反映流行区钉螺的血吸虫感染状态。
Objective To investigate the difference of detection rate of snail in different stages of schistosomiasis infection by ring-mediated simultaneous amplification of DNA (LAMP) and dissection-microscopy. Methods Infected with Schistosoma japonicum miracidia, cultured in a 25 ℃ incubator. Ten experimental snails were collected at 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 weeks after infection, respectively. Genomic DNA was extracted from the dissected snail and detected by LAMP method. At 10 weeks after infection, 100 experimental snails were obtained and simultaneously examined by dissection-microscopy and LAMP. Collect snail snail endemic areas on the scene, using two methods to detect at the same time, statistics of schistosomiasis infection snail positive rate. Results The infection rates of 10 snails of T. gonorrhoeae were 0.00%, 0.00%, 0.00%, 10.00%, 0.00,10.00%, 20.00%, 20.00% and 60.00%, respectively, from 1 to 10 weeks after the dissection-microscopy and LAMP assay. 60.00%, 50.00%, 20.00%, 30.00%, 20.00%, 20.00%, 40.00%, 30.00%, 40.00%, 80.00%, 70.00% respectively. The total positive rates were 18.00% and 40.00% χ2 = 11.75, P <0.01). The positive rate of 100 snails in the seven weeks after infection was 31.00% and 39.00% respectively. The positive rates of snails from the endemic areas of schistosomiasis in two different provinces were 29.80% and 19.47% respectively. Conclusion The ring-mediated isothermal DNA amplification method can effectively detect infectious snails at different stages of infection, and the detection results can more accurately reflect the infection status of schistosomiasis in the endemic snails.