Gallas-Lindemann Carmen, Sotiriadou Isaia, Plutzer Judit, Noack Michael J, Mahmoudi Mohammad Reza, Karanis Panagiotis
Medical and Molecular Parasitology Laboratory, University of Cologne, Medical School, Center of Anatomy, Institute II, Cologne, Germany; Linksniederrheinische Entwässerungs-Genossenschaft, Friedrich-Heinrich-Allee 64, 47475, Kamp-Lintfort, Germany.
Medical and Molecular Parasitology Laboratory, University of Cologne, Medical School, Center of Anatomy, Institute II, Cologne, Germany; Centre of Dental Medicine, Policlinic of Operative Dentistry and Periodontology, University of Cologne, Germany.
Acta Trop. 2016 Jun;158:43-51. doi: 10.1016/j.actatropica.2016.02.005. Epub 2016 Feb 12.
Environmental water samples from the Lower Rhine area in Germany were investigated via immunofluorescence assays (IFAs), nested polymerase chain reaction (nested PCR) and loop-mediated isothermal amplification (LAMP) to detect the presence of Giardia spp. (n=185) and Cryptosporidium spp. (n=227). The samples were concentrated through filtration or flocculation, and oocysts were purified via centrifugation through a sucrose density gradient. For all samples, IFA was performed first, followed by DNA extraction for the nested PCR and LAMP assays. Giardia cysts were detected in 105 samples (56.8%) by IFA, 62 samples (33.5%) by nested PCR and 79 samples (42.7%) by LAMP. Cryptosporidium spp. were detected in 69 samples (30.4%) by IFA, 95 samples (41.9%) by nested PCR and 99 samples (43.6%) by LAMP. According to these results, the three detection methods are complementary for monitoring Giardia and Cryptosporidium in environmental waters.
通过免疫荧光分析(IFA)、巢式聚合酶链反应(巢式PCR)和环介导等温扩增(LAMP)对德国莱茵河下游地区的环境水样进行检测,以确定贾第虫属(n = 185)和隐孢子虫属(n = 227)的存在情况。通过过滤或絮凝对样品进行浓缩,然后通过蔗糖密度梯度离心法纯化卵囊。对于所有样品,首先进行IFA检测,随后提取DNA用于巢式PCR和LAMP检测。通过IFA在105份样品(56.8%)中检测到贾第虫包囊,通过巢式PCR在62份样品(33.5%)中检测到,通过LAMP在79份样品(42.7%)中检测到。通过IFA在69份样品(30.4%)中检测到隐孢子虫属,通过巢式PCR在95份样品(41.9%)中检测到,通过LAMP在99份样品(43.6%)中检测到。根据这些结果,这三种检测方法在监测环境水体中的贾第虫和隐孢子虫时具有互补性。