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用于食品和牛粪中大肠杆菌O157检测的快速疏水网格膜过滤酶标抗体法的灵敏度提高。

Increased sensitivity of the rapid hydrophobic grid membrane filter enzyme-labeled antibody procedure for Escherichia coli O157 detection in foods and bovine feces.

作者信息

Szabo R, Todd E, MacKenzie J, Parrington L, Armstrong A

机构信息

Bureau of Microbial Hazards, Health Protection Branch, Tunney's Pasture, Ottawa, Ontario, Canada.

出版信息

Appl Environ Microbiol. 1990 Nov;56(11):3546-9. doi: 10.1128/aem.56.11.3546-3549.1990.

Abstract

Several strains of Escherichia coli O157:H7 artificially inoculated into vegetables and dairy products were recovered on hydrophobic grid membrane filters and enumerated by an enzyme-labeled antibody assay. The mean of the recoveries from 12 fresh vegetables was 108.8%, whereas that from 10 dairy products was 93.2%. Modified tryptic soy broth at 43 degrees C with shaking at 100 rpm provided optimum recovery of the organism from meat, with a sensitivity of less than or equal to 1 CFU/g, which is 10 times more sensitive than direct plating. The method performed equally well with vegetable and dairy products. Tryptic soy broth, however, under the same conditions gave the best results for fecal samples. Of 22 asymptomatic dairy cattle, reported as having positive Brucella titers when assayed with polyclonal antibodies, eight were found to contain E. coli O157 in their feces as demonstrated by the enzyme-labeled antibody assay by using monoclonal antibodies. This finding may explain some of the false-positive Brucella tests.

摘要

将几株大肠杆菌O157:H7人工接种到蔬菜和乳制品中,然后用疏水网格膜过滤器进行回收,并通过酶标抗体检测法进行计数。12种新鲜蔬菜的回收率平均值为108.8%,而10种乳制品的回收率平均值为93.2%。43℃的改良胰蛋白胨大豆肉汤,以100 rpm振荡,能从肉类中最佳回收该菌,灵敏度小于或等于1 CFU/g,比直接平板计数法灵敏10倍。该方法对蔬菜和乳制品同样有效。然而,在相同条件下,胰蛋白胨大豆肉汤对粪便样本的检测效果最佳。在22头无症状奶牛中,用多克隆抗体检测时布鲁氏菌滴度呈阳性,通过使用单克隆抗体的酶标抗体检测法发现其中8头奶牛的粪便中含有大肠杆菌O157。这一发现可能解释了一些布鲁氏菌检测假阳性的情况。

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Monoclonal antibodies for detection of the H7 antigen of Escherichia coli.用于检测大肠杆菌H7抗原的单克隆抗体。
Appl Environ Microbiol. 1996 Sep;62(9):3325-32. doi: 10.1128/aem.62.9.3325-3332.1996.

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