Investigation of Coxiella burnetii contamination in commercial milk and PCR method for the detection of C. burnetii in egg |
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Authors: | Hirai Akihiko Kaneko Seiji Nakama Akiko Ishizaki Naoto Odagiri Megumi Kai Akemi Sadamasu Kenji Shinkai Takayuki Yano Kazuyoshi Morozumi Satoshi |
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Affiliation: | Tokyo Metropolitan Institute of Public Health: 3-24-1, Hyakunincho, Shinjuku-ku, Tokyo 169-0073, Japan. |
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Abstract: | A total of 244 milk samples collected from supermarkets in Tokyo were examined for contamination with Coxiella burnetii. C. burnetii DNA was detected in 131 (53.7%) of the samples by nested PCR. PCR-positive samples were injected into immunosuppressed A/J strain mice. Of the 22 PCR-positive milk samples tested, none resulted in isolation of C. burnetii from the mice. Heat-treatment was sufficient to inactivate C. burnetii in commercial milk. In addition, a PCR detection method for C. burnetii in chicken egg was developed. Egg yolk was added to an equal volume of 1 mol/L of NaCl phosphate buffer and homogenized for removal of protein and lipid. After centrifugal separation, the supernatant was removed, and template DNA in the precipitate was extracted using SDS, proteinase K and NaI. Using such prepared samples, 3.2 x 10(1) C. burnetii particles in 1 g of egg yolk could be detected by nested PCR. All of 200 chicken egg samples collected from supermarkets in Tokyo were negative for C. burnetii by the nested PCR method. |
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