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目的用PCR和免疫荧光染色镜检法检测城市现场水源中蓝氏贾第鞭毛虫(简称贾第虫)和隐孢子虫,了解上海市水源地贾第虫和隐孢子虫污染状况。方法采集上海市8个区的自来水和3个区的环境水,PCR检测水样本中的贾第虫磷酸丙糖异构酶(triosephosphate isomerase,Tim)基因和隐孢子虫18S rRNA的基因;按《生活饮用水标准检验方法》(GB5750.12-2006)Filta-Max Xpress快速方法进行免疫荧光染色镜检。计算比较两种方法检测贾第虫和隐孢子虫的阳性率。结果共采集200份水样;其中自来水出厂水48份,PCR和荧光镜检法均未检出贾第虫和隐孢子虫;检测原水、动物饲养场周边池水、污水处理厂出水、游泳池水和餐馆养渔池等水样分别为62、25、29、20、16份,PCR法贾第虫阳性率分别为8.1%、36.0%、17.2%、0和0,总阳性率为12.5%;隐孢子虫阳性率分别为6.5%、40.0%、13.8%、0和0,总阳性率为11.8%。免疫荧光染色镜检法贾第虫阳性率分别为9.7%、40.0%、24.1%、0和0,总阳性率为15.1%;隐孢子虫阳性率分别为8.1%、44.0%、17.2%、0和0,总阳性率为13.8%。两种方法检测贾第虫和隐孢子虫一致性分别为96.1%和95.4%,经Kappa检验,两种方法一致性较好(Kappa=0.83和0.79,Kappa>0.75),PCR与荧光镜检结果无显著性差异(χ~2=0.44和0.26,P<0.05)。结论两种方法均可用于城市现场水源中贾第虫和隐孢子虫污染的调查。自来水未检出贾第虫和隐孢子虫,但在原水和环境水中检测到贾第虫和隐孢子虫污染,需加强监测。
Objective To detect Giardia lamblia (Giardia lamblia) and Cryptosporidium parvum in urban water sources by PCR and immunofluorescence staining, and to know the status of Giardia and Cryptosporidium in water sources in Shanghai. Methods The tap water and the environmental water in three districts were collected from 8 districts of Shanghai. PCR was used to detect the gene of Timothyria triosephosphate isomerase (Tim) and Cryptosporidium 18S rRNA in water samples. According to “ Standard Test Method for Drinking Water ”(GB5750.12-2006) Filta-Max Xpress Fast method for immunofluorescence microscopy. The positive rates of Giardia and Cryptosporidium were calculated and compared by two methods. Results A total of 200 water samples were collected. Among them, 48 samples were made from tap water, no Giardia and Cryptosporidium were detected by PCR and fluorescence microscopy. Water samples were collected from raw water, feedwater around ponds, effluent from sewage treatment plants, The water samples in the rearing ponds of restaurants were 62, 25, 29, 20 and 16 respectively. The positive rates of Giardia PCR were 8.1%, 36.0%, 17.2%, 0 and 0, respectively. The overall positive rate was 12.5% The positive rates of sporozoites were 6.5%, 40.0%, 13.8%, 0 and 0, respectively. The total positive rate was 11.8%. The positive rate of Giardia was 9.7%, 40.0%, 24.1%, 0 and 0, respectively. The positive rate was 15.1%. The positive rates of Cryptosporidium were 8.1%, 44.0%, 17.2%, 0 And 0, the total positive rate was 13.8%. The consistency of the two methods for detecting Giardia and Cryptosporidium were 96.1% and 95.4%, respectively. The Kappa test showed that the two methods had good agreement (Kappa = 0.83 and 0.79, Kappa> 0.75), PCR and fluorescence microscopy No significant difference (χ ~ 2 = 0.44 and 0.26, P <0.05). Conclusion Both methods can be used to investigate Giardia and Cryptosporidium contamination in urban water sources. Giardia and Cryptosporidium were not detected in tap water, but Giardia and Cryptosporidium contamination was detected in raw and ambient water and monitoring was warranted.