Food Safety Key Laboratory of Zhejiang Province, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310035, People's Republic of China.
Curr Microbiol. 2011 May;62(5):1423-30. doi: 10.1007/s00284-011-9870-z. Epub 2011 Jan 30.
The rep-PCR DNA fingerprinting performed with REP, BOX A1R, and (GTG)(5) primers was investigated as a way to differentiate between human, livestock, and poultry sources of fecal pollution on the area of Xiangshan Bay, East China Sea. Of the three methods, the BOX-PCR DNA fingerprints analyzed by jack-knife algorithm were revealed high rate of correct classification (RCC) with 91.30, 80.39, 89.39, 86.14, 93.24, 87.72, and 89.28% of human, cattle, swine, chicken, duck, sheep, and goose E. coli isolates classified into the correct host source, respectively. The average rate of correct classification (ARCC) of REP-, BOX-, and (GTG)(5)-PCR patterns was 79.88, 88.21, and 86.39%, respectively. Although the highest amount of bands in (GTG)(5)-PCR fingerprints could be observed, the discriminatory efficacy of BOX-PCR was superior to both REP- and (GTG)(5)-PCR. Moreover, the similarity of 459 isolates originated from shellfish and growing water was compared with fecal-obtained strains. The results showed that 92.4 and 96.2% E. coli strains isolated from midstream and downstream shellfish samples, respectively, had a ≥ 80% similarity with corresponding strains isolated from fecal samples. It was indicated that E. coli in feces could spread from human sewage or domestic farms to the surrounding shellfish culture water, and potentially affect the quality of shellfish. This work suggests that rep-PCR fingerprinting can be a promising genotypic tool applied in the shellfish growing water management on East China Sea for source identification of fecal pollution.
REP、BOX A1R 和 (GTG)(5) 引物的重复扩增多态性 DNA 指纹图谱分析被用来区分中国东海象山湾地区人类、家畜和家禽来源的粪便污染。在这三种方法中，通过 Jackknife 算法分析的 BOX-PCR DNA 指纹图谱显示出较高的正确分类率（RCC），分别有 91.30%、80.39%、89.39%、86.14%、93.24%、87.72%和 89.28%的人类、牛、猪、鸡、鸭、羊和鹅大肠杆菌分离株被正确分类到宿主来源。REP、BOX 和 (GTG)(5)-PCR 模式的平均正确分类率（ARCC）分别为 79.88%、88.21%和 86.39%。尽管 (GTG)(5)-PCR 指纹图谱中可以观察到最多的条带，但 BOX-PCR 的区分效果优于 REP-PCR 和 (GTG)(5)-PCR。此外，还比较了贝类和养殖水中的 459 株分离株与粪便获得的菌株之间的相似性。结果表明，从中游和下游贝类样本中分离的 92.4%和 96.2%的大肠杆菌菌株与从粪便样本中分离的相应菌株的相似性≥80%。这表明粪便中的大肠杆菌可能会从人类污水或家庭农场传播到周围的贝类养殖水中，并可能影响贝类的质量。这项工作表明，重复扩增多态性 DNA 指纹图谱分析可以作为一种有前途的基因型工具，应用于中国东海贝类养殖水管理中的粪便污染源识别。
