Sadamasu Kenji, Tabei Yukiko, Shinkai Takayuki, Hasegawa Michiya, Kaneko Seiji, Hirai Akihiko, Nakama Akiko, Ishizaki Naoto, Odagiri Megumi, Kamata Shin-ichi, Yano Kazuyoshi, Kai Akemi, Morozumi Satoshi
Department of Microbiology, Tokyo Metropolitan Institute of Public Health, 3-24-1, Shinjuku-ku, Tokyo 169-0073, Japan.
Shokuhin Eiseigaku Zasshi. 2006 Feb;47(1):1-8.
A PCR method for the effective detection of Coxiella burnetii in commercially available mayonnaise was developed. Sample preparations were isolated from 50 g portions of each mayonnaise product by four successive extraction steps in phosphate buffer with 2.0 M NaCl. These extracts were then centrifuged at 20,000 x g for 60 min. DNA was isolated from the solution containing the precipitate with a commercial kit, and amplified quantitatively using real-time PCR that targeted the com1 region of C. burnetii. The recoveries of C. burnetii from 2 kinds of commercial mayonnaise specimens, with a baseline control of 1 x 10(7) particles of the Nine Mile phase II strain, were 85.0 +/- 6.0% and 72.0 +/- 0.4%, respectively. The determination limit of this method was 500 C. burnetii particles per 50 g of mayonnaise. The DNA specimens isolated from 50 different commercial mayonnaise samples sold in Tokyo using this method were amplified using both nested PCR and real-time PCR. No contamination by C. burnetii was detected in any of the mayonnaise samples.
开发了一种用于有效检测市售蛋黄酱中伯氏考克斯体的聚合酶链反应（PCR）方法。通过在含有2.0 M氯化钠的磷酸盐缓冲液中进行四个连续提取步骤，从每份50 g的蛋黄酱产品中分离样品制剂。然后将这些提取物在20,000×g下离心60分钟。使用商业试剂盒从含有沉淀物的溶液中分离DNA，并使用针对伯氏考克斯体com1区域的实时PCR进行定量扩增。以九英里II期菌株1×10⁷个颗粒作为基线对照，从两种市售蛋黄酱标本中回收伯氏考克斯体的回收率分别为85.0±6.0%和72.0±0.4%。该方法的检测限为每50 g蛋黄酱中500个伯氏考克斯体颗粒。使用该方法从东京销售的50个不同市售蛋黄酱样品中分离的DNA标本，通过巢式PCR和实时PCR进行扩增。在任何蛋黄酱样品中均未检测到伯氏考克斯体污染。
