United States Environmental Protection Agency, Office of Research and Development, USA.
United States Environmental Protection Agency, Office of Research and Development, USA.
J Virol Methods. 2021 Oct;296:114245. doi: 10.1016/j.jviromet.2021.114245. Epub 2021 Jul 24.
Dead-end hollow fiber ultrafiltration combined with a single agar layer assay (D-HFUF-SAL) has potential use in the assessment of sanitary quality of recreational waters through enumeration of coliphage counts as measures of fecal contamination. However, information on applicability across a broad range of sites and water types is limited. Here, we tested the performance of D-HFUF-SAL on 49 marine and freshwater samples. Effect of method used to titer the spiking suspension (SAL versus double agar layer [DAL]) on percent recovery was also evaluated. Average somatic coliphage recovery (72 % ± 27) was significantly higher (p < 0.0001) compared to F+ (53 % ± 19). This was more pronounced for marine (p ≤ 0.0001) compared to freshwaters (p = 0.0134). Neither method affected somatic coliphage, but DAL (28 % ± 12) significantly (p < 0.0001) underestimated F + coliphage recoveries compared to SAL (53 % ± 19). Overall, results indicate that, while D-HFUF-SAL performed well over a wide variety of water types, F + coliphage recoveries were significantly reduced for marine waters suggesting that some components unique to this habitat may interfere with the assay performance. More importantly, our findings indicate that choice of spike titer method merits careful consideration since it may under-estimate method percent recovery.
死端中空纤维超滤结合单层琼脂层检测法(D-HFUF-SAL)在通过噬菌体计数评估休闲水中卫生质量方面具有潜在用途,噬菌体计数可作为粪便污染的指标。然而,关于该方法在广泛的地点和水类型中适用性的信息有限。在此,我们用 49 个海水和淡水样本对 D-HFUF-SAL 进行了测试。还评估了用于滴定接种悬浮液的方法(SAL 与双层琼脂层[DAL])对回收率的影响。与 F+(53%±19)相比,平均体细胞噬菌体回收率(72%±27)明显更高(p<0.0001)。与淡水相比,这种情况在海水中更为明显(p≤0.0001)。两种方法都不会影响体细胞噬菌体,但与 SAL(53%±19)相比,DAL(28%±12)显著(p<0.0001)低估了 F+噬菌体的回收率。总的来说,结果表明,尽管 D-HFUF-SAL 在各种水类型中表现良好,但由于海洋水中某些特有成分可能干扰检测性能,F+噬菌体的回收率显著降低。更重要的是,我们的研究结果表明,由于选择了滴定方法,因此需要仔细考虑,因为它可能会低估方法的回收率。