Sciacchitano C J, Hirshfield I N
U.S. Food and Drug Administration, Northeast Regional Laboratory, Brooklyn, NY 11232, USA.
J AOAC Int. 1996 Jul-Aug;79(4):861-5.
The polymerase chain reaction (PCR), a rapid, sensitive technique for amplifying target DNA sequences of pathogenic microorganisms, was used to amplify Clostridium botulinum type E neurotoxin gene fragments in smoked fish. Other botulinal neurotoxin-producing strains, nontoxigenic strains, and food-related microorganisms did not yield nonspecific amplification products with this PCR assay. PCR products were analyzed by capillary electrophoresis (CE) using a low-viscosity entangled polymer system. Resolution, sensitivity, and DNA sizing accuracy were improved, and analytical times were markedly shortened. The PCR/CE assay detected the C. botulinum type E neurotoxin gene in as few as 10 cells. The technique to other foods may also be a valuable tool for detecting foodborne pathogens.
聚合酶链反应（PCR）是一种用于扩增致病微生物靶DNA序列的快速、灵敏技术，被用于扩增烟熏鱼中肉毒杆菌E型神经毒素基因片段。其他产肉毒杆菌神经毒素的菌株、无毒菌株以及与食品相关的微生物在该PCR检测中均未产生非特异性扩增产物。使用低粘度缠结聚合物系统通过毛细管电泳（CE）对PCR产物进行分析。分辨率、灵敏度和DNA大小测定准确性均得到提高，分析时间显著缩短。该PCR/CE检测方法可检测低至10个细胞中的肉毒杆菌E型神经毒素基因。该技术对于其他食品而言，也可能是检测食源性病原体的一种有价值的工具。
