以巢式聚合酶連鎖反應及核酸序列分析檢測貓感染艾利希體（Ehrlichia canis）

Identification of Ehrlichi Canis in Cats by Nested Polymerase Chain Reaction and Nucleotide Sequence Analysis

殷嘉諄(Chia-Chun Yin) ； 劉宏仁(Hung-Jen Liu) ； 林孫權(Suen-Chuain Lin) ； 廖明輝(Ming-Huei Liao) ； 吳永惠(Yeong-Huey Wu)

《臺灣獸醫學雜誌》 29卷2期 (2003/06) Pp. 122-128

https://doi.org/10.7009/TVJ.200306.0122

摘要

本研究以巢式聚合酶連鎖反應及核酸序列分析證實台灣地區貓隻感染Ehrlichia canis。在1999年6月至2000年3月間，收集國立屏東科技大學附設教學動物醫院門診貓病例及高雄市開業獸醫院臨床病例，血液樣本以血液學檢查及血清學IFA檢測感染於貓之艾利希體，調查貓感染艾利希體之情形。在血液學檢查方面，所有貓隻病例皆無法在末梢血液中見到細胞質含偶氮顆粒樣包涵體之淋巴球或單核球，但可見貧血及雙血球減少症。在IFA抗體測定則皆呈陰性反應。以巢式PCR檢測17個病貓病例，陽性反應率為11.76%（2/17）。以DNASTART軟體比對樣品核酸序列及已發表序列之相似與相異度，結果顯示相似度高達99.4%以上，證實PCR產物皆為Ehrlichia canis之16S rRNA基因。

關鍵字

艾利希體 ； 巢式聚合酶連鎖反應 ； 間接螢光抗體試驗

並列摘要

Ehrlichia canis infection in cats was identified by nested polymerase chain reaction (PCR) and nucleotide sequence analysis. Feline blood samples were collected from clinical cases at Teaching Animal Hospital, National Pingtung University of Science and Technology and from clinical samples in Kaohsiung city in June, 1999 to March, 2000. In the present study, blood samples were examined by blood smear examination and an indirect fluorescent-antibody (IFA) test for detecting Ehrlichia canis. In blood smear examination, Ehrlichial inclusion was not seen in peripherallymphocytes or monocytes of all cats, but bicytopenia and anemia were found in PCR-positive cats. Two out of 17 cats (11.76%) showed positive with nested PCR, while all of these cats were IFA negative. To further confirm PCR, PCR products were sequenced and compared with previously published sequences by DNASTART software. The sequence similarities of 16S rRNA gene among them were above 99.4 %. Sequence analysis confirmed that all of them were Ehrlichia canis.

並列關鍵字

Ehrlichi ； Indirect fluorescent-antibody (IFA) test ； Nested polymerase chain reaction (PCR)