中国环境科学
中國環境科學
중국배경과학
CHINA ENVIRONMENTAL SCIENCE
2014年
8期
2118-2125
,共8页
王海鹰%贾乐华%吴仁人%王菊芳%宁正祥
王海鷹%賈樂華%吳仁人%王菊芳%寧正祥
왕해응%가악화%오인인%왕국방%저정상
拟杆菌%特异性引物%灵敏度%特异性%珠江三角洲地区
擬桿菌%特異性引物%靈敏度%特異性%珠江三角洲地區
의간균%특이성인물%령민도%특이성%주강삼각주지구
Bacteroidales%specific markers%sensitivity%specificity%Pearl River Delta region
在珠江三角洲地区采集人、猪、牛、狗以及家禽等33个粪便样品,高效提取基因组DNA,选取14种特异性拟杆菌引物进行检验分析,并进一步采用已知污染源类型的水样对其进行验证。结果表明：采用粪便样品DNA专属提取试剂盒和水体样品DNA专属提取试剂盒提取的基因组模版纯度和提取率均符合后续实验要求。拟杆菌通用引物2#(Bac32F/Bac708R)检出率为85%(28/33),特别是对哺乳类动物和鸡的粪便具有更高的检出率。人的拟杆菌特异性引物3#(HF134F/ Bac708R)和4#(HF183F/Bac708R)、反刍动物的拟杆菌特异性引物8#(CF128F/Bac708R)以及猪的拟杆菌特异性引物10#(PF163F/Bac708R)在珠江三角洲地区同时具有较高的灵敏度和较强的特异性。水体样品验证实验与实际污染类型相符,说明拟杆菌通用引物2#、人的特异性引物3#和4#以及猪的特异性引物10#在珠江三角洲地区具有很好的适用性。
在珠江三角洲地區採集人、豬、牛、狗以及傢禽等33箇糞便樣品,高效提取基因組DNA,選取14種特異性擬桿菌引物進行檢驗分析,併進一步採用已知汙染源類型的水樣對其進行驗證。結果錶明：採用糞便樣品DNA專屬提取試劑盒和水體樣品DNA專屬提取試劑盒提取的基因組模版純度和提取率均符閤後續實驗要求。擬桿菌通用引物2#(Bac32F/Bac708R)檢齣率為85%(28/33),特彆是對哺乳類動物和鷄的糞便具有更高的檢齣率。人的擬桿菌特異性引物3#(HF134F/ Bac708R)和4#(HF183F/Bac708R)、反芻動物的擬桿菌特異性引物8#(CF128F/Bac708R)以及豬的擬桿菌特異性引物10#(PF163F/Bac708R)在珠江三角洲地區同時具有較高的靈敏度和較彊的特異性。水體樣品驗證實驗與實際汙染類型相符,說明擬桿菌通用引物2#、人的特異性引物3#和4#以及豬的特異性引物10#在珠江三角洲地區具有很好的適用性。
재주강삼각주지구채집인、저、우、구이급가금등33개분편양품,고효제취기인조DNA,선취14충특이성의간균인물진행검험분석,병진일보채용이지오염원류형적수양대기진행험증。결과표명：채용분편양품DNA전속제취시제합화수체양품DNA전속제취시제합제취적기인조모판순도화제취솔균부합후속실험요구。의간균통용인물2#(Bac32F/Bac708R)검출솔위85%(28/33),특별시대포유류동물화계적분편구유경고적검출솔。인적의간균특이성인물3#(HF134F/ Bac708R)화4#(HF183F/Bac708R)、반추동물적의간균특이성인물8#(CF128F/Bac708R)이급저적의간균특이성인물10#(PF163F/Bac708R)재주강삼각주지구동시구유교고적령민도화교강적특이성。수체양품험증실험여실제오염류형상부,설명의간균통용인물2#、인적특이성인물3#화4#이급저적특이성인물10#재주강삼각주지구구유흔호적괄용성。
A total of 33 fecal samples of human and different animal species were collected in the Pearl River Delta region. Genomic DNA were then efficiently extracted and amplified by using 14 specific Bacteroidales primers that have been reported as tracking markers of different sources. Primers with high specificity and sensitivity were then selected and used for polluted water samples analysis. The results show that the purity and extract efficiency of the genome DNA meet the requirement of the following experiments. 85%of the 33 fecal samples analyzed were positive when using Bacteroidales universal primer 2#(Bac32F/Bac708R), and most of the mammalian animals and chicken fecal samples were detected as positive samples when use this primer. Human-specific marker 3#(HF134F/Bac708R) and 4#(HF183F/Bac708R), ruminant-specific marker 8#(CF128F /Bac708R)and porcine-specific marker 10#(PF163F/Bac708R)showed high sensitivity and specificity in the Pearl River Delta region. Polluted water samples analysis results were the same as the actual type of pollution, implying Bacteroidales universal primer 2#, human-specific primer 3# and 4# and porcine-specific primer 10# could be used as pollution tracking markers in the Pearl River Delta region.
