Koch W H, Payne W L, Wentz B A, Cebula T A
Division of Microbiology, Food and Drug Administration, Washington, D.C. 20204.
Appl Environ Microbiol. 1993 Feb;59(2):556-60. doi: 10.1128/aem.59.2.556-560.1993.
The polymerase chain reaction was used to selectively amplify sequences within the cholera toxin operon from Vibrio cholerae O1. Oysters, crabmeat, shrimp, and lettuce were seeded with V. cholerae and then homogenized or washed with alkaline peptone water, followed by short-term (6- to 8-h) enrichment. A detection limit of as few as 1 V. cholerae CFU per 10 g of food was obtained with amplification reactions from crude bacterial lysates. The method is extremely rapid and obviates the need for DNA isolation from a variety of complex food matrices.
采用聚合酶链反应从霍乱弧菌O1中选择性扩增霍乱毒素操纵子内的序列。将霍乱弧菌接种到牡蛎、蟹肉、虾和生菜中，然后用碱性蛋白胨水匀浆或冲洗，接着进行短期（6至8小时）富集培养。通过对粗制细菌裂解物进行扩增反应，获得了每10克食物中低至1个霍乱弧菌CFU的检测限。该方法极其快速，无需从各种复杂的食物基质中分离DNA。
