Liu Yongcheng, Ye Jianming, Li Yanbin
Department of Biological and Agricultural Engineering, Center of Excellence for Poultry Science, University of Arkansas, POSC O-411, Fayetteville, Arkansas 72701, USA.
J Food Prot. 2003 Mar;66(3):512-7. doi: 10.4315/0362-028x-66.3.512.
A biosensor was evaluated with regard to its usefulness in the rapid detection of Escherichia coli O157:H7 inoculated in ground beef, chicken carcass, and romaine lettuce samples. The biosensor consisted of a chemiluminescence reaction cell, a fiber-optic light guide, and a luminometer linked to a personal computer in conjunction with immunomagnetic separation. The samples inoculated with E. coli O157:H7 were first centrifuged and suspended in buffered peptone water and then incubated with anti-E. coli O157 antibody-coated magnetic beads and horseradish peroxidase (HRP)-labeled anti-E. coli O157 antibodies to form antibody-coated bead-bacterium-HRP-labeled antibody sandwich complexes. Finally, the sandwich complexes were separated from the samples in a magnetic field and reacted with luminol in the reaction cell. The number of E. coli O157:H7 cells was determined by collecting the HRP-catalyzed chemiluminescence signal from the bead surface through a fiber-optic light guide and measuring the signal with a luminometer. The chemiluminescence biosensor was specific for E. coli O157:H7 in samples containing other bacteria, including Salmonella Typhimurium, Campylobacter jejuni, and Listeria monocytogenes. The chemiluminescence signal was linear on a log scale from 10(2) to 10(5) CFU of E. coli O157:H7 per ml in samples. Detection could be completed within 1.5 h without any enrichment. The detection limits for ground beef, chicken carcass, and lettuce samples were 3.2 x 10(2), 4.4 x 10(2), and 5.5 x 10(2) CFU of E. coli O157:H7 per ml, respectively.
对一种生物传感器在快速检测接种于绞碎牛肉、鸡胴体和生菜样本中的大肠杆菌O157:H7方面的实用性进行了评估。该生物传感器由一个化学发光反应池、一个光纤光导和一台与个人计算机相连的发光计组成,并结合了免疫磁分离技术。接种了大肠杆菌O157:H7的样本首先进行离心,然后悬浮于缓冲蛋白胨水中,接着与抗大肠杆菌O157抗体包被的磁珠以及辣根过氧化物酶(HRP)标记的抗大肠杆菌O157抗体一起孵育,以形成抗体包被磁珠-细菌-HRP标记抗体夹心复合物。最后,在磁场中将夹心复合物与样本分离,并在反应池中与鲁米诺反应。通过光纤光导从磁珠表面收集HRP催化的化学发光信号,并用发光计测量该信号,从而确定大肠杆菌O157:H7细胞的数量。该化学发光生物传感器对含有其他细菌(包括鼠伤寒沙门氏菌、空肠弯曲菌和单核细胞增生李斯特菌)的样本中的大肠杆菌O157:H7具有特异性。在样本中,每毫升大肠杆菌O157:H7的化学发光信号在10(2)至10(5) CFU的对数范围内呈线性。无需任何富集,检测可在1.5小时内完成。绞碎牛肉、鸡胴体和生菜样本的检测限分别为每毫升3.2 x 10(2)、4.4 x 10(2)和5.5 x 10(2) CFU的大肠杆菌O157:H7。