@article{oai:repository.naro.go.jp:00001772,
 author = {猪島, 康雄 and INOSHIMA, Yasuo and 今井, 邦俊 and IMAI, Kunitoshi and 村上, 賢二 and MURAKAMI, Kenji and 西森, 知子 and NISHIMORI, Tomoko and 清水, 眞也 and SHIMIZU, Shinya and 横山, 隆 and YOKOYAMA, Takashi and 泉對, 博 and SENTSUI, Hiroshi},
 journal = {動物衛生研究所研究報告, Bulletin of the National Institute of Animal Health},
 month = {Mar},
 note = {Bovine mammillitis has been sporadically observed among cattle in Japan. Since parapoxvirus (PPV) and bovine herpesvirus type 2 (BHV-2) were suspected as agents of the diseases, specific and sensitive methods for diagnosis of these diseases were studied. Seroepidemiological surveys were performed on PPV by agar gel immunodiffusion test and ELISA using protein AG, and on BHV-2 by virus neutralization tests. Serum samples were collected from cattle in various parts of Japan. Positive reactors to PPV were found all over Japan and the rate of positive reactors was more than 70%. On the other hand, reactors to BHV-2 were found sporadically and their positive rate was about 1%. Positive reactors to PPV were observed in Japanese serow, suggesting that PPV is transmitted between domestic and wild animals. Genetical diagnosis methods by PCR were developed for the detection of PPV and BHV-2, and their practical use was evaluated. Semi-nested PCR was applied to the diagnosis of PPV infection and it was able to detect viral DNA from lesions of clinically affected animals in the fields. The reaction was effective for all PPV strains and these were divided into 4 groups by restriction fragment length polymorphism. Shuttle PCR was developed for diagnosis of BHV-2 infection and the specificity and sensitivity of the reaction were confirmed by digestion with restriction endonuclease AulI and by using homogenized muscle and skin of mice experimentally infected with BHV-2., 日本各地で散発的に発生している牛の乳頭炎の病因として, パラポックスウイルス（PPV)および牛ヘルペスウイルス2型（BHV-2)が疑われている. これらウイルス性疾病の特異的診断法の開発を試みるとともに, 国内における浸潤状況や生態系の調査を行った. PPVは寒天ゲル内沈降試験およびプロテインAGを使用したELISAで, BHV-2は中和試験で, 全国各地から集めた牛血清の抗体検査を行った. PPV抗体陽性牛は広範囲に存在し, 陽性率は70％以上であった. 一方, BHV-2の抗体陽性牛の分布は限局しており, 陽性率は約1％であった. 野生動物ではニホンカモシカで高率にPPV抗体陽性例が検出され, 家畜との間で本ウイルスの生態系が形成されている可能性が示唆された. 両ウイルスについてPCRによる遺伝子診断法を開発し, その実用性を検討した. PPVでは, 全ての種に共通なPCRプライマ－を設定し, 野外における発症動物の病変組織から, 直接遺伝子診断を行う手法を開発した. また, そのPCR産物の制限酵素切断像から, 4種類のウイルス分類を可能とした. BHV-2では, 実験感染した乳のみマウスの筋肉・皮膚乳剤を用いて, shuttle PCR と増幅部位のAulI切断像から, 高感度で特異的診断が行えることを明らかにした.},
 pages = {23--32},
 title = {牛ウイルス性乳頭炎の調査および診断法の開発},
 volume = {108},
 year = {2002},
 yomi = {イノシマ, ヤスオ and イマイ, クニトシ and ムラカミ, ケンジ and ニシモリ, トモコ and シミズ, シンヤ and ヨコヤマ, タカシ and センツイ, ヒロシ}
}