Two-Step Polymerase Chain Reaction for Detection of Ehrlichia Platys in Canine Infectious Cyclic Thrombocytopenia

應用兩階段聚合酶鏈反應診斷犬傳染性週期性血小板缺乏症

張溫嵐(Wen-Lan Chang) ； 潘銘正(Ming-Jeng Pan)

《中華民國獸醫學會雜誌》 22卷1期 (1996/02) Pp. 9-16

https://doi.org/10.7009/JTCSVS.199602.0009

摘要

犬傳染性週期性血小板缺乏症之診斷主要是籍由觀察染色血液抹片中存在血小板內之嗜鹼性包涵體。然而，以這個方法診斷本病並不十分可靠，特別是在慢性感染階段。本報告利用兩階段聚合酶鏈反應偵測引起犬傳染性週期性血小板缺乏症之病原。血小板艾利希氏體之16S核醣體RNA基因中之二區域被選擇做為核酸增幅之目標，所有的增幅反應均是採用快速溫度循環系統來進行。將此方法應用於由台大家畜醫院收集到之10個血小板嚴重低於正常值之血液病材，其中有3個呈陽性反應，而當中只有1個可於染色之血液抹片中觀察到包涵體。由結果看來，應用兩階段聚合酶鏈反應於本病之臨床診斷，其敏感性、專一性和時效性均相當高。

關鍵字

犬傳染性週期性血小板缺乏症 ； 血小板艾利希氏體 ； 聚合酶鏈反應

並列摘要

Canine infectious cyclic thrombocytopenia (CICT) is diagnosed by finding the basophilic inclusions within platelets on stained blood films. However, this method is unreliable, especially for detection at the chronic stage of infection. This report describes a two-step polymerase chain reaction (PCR) for detection of the etiologic agent of CICT. Two regions of the 16S rRNA gene sequence were used to amplify DNA specifically from Ehrlichia platys in blood specimens. All amplification reactions were run with the standard rapid cycling system. This technique was applied to 10 severely thrombocytopenic blood specimens collected from National Taiwan University Veterinary Hospital. Three of them were positive by two-step PCR amplification while only one is positive by Giemsa stained blood smear observation. The results indicate that this two-step PCR is a highly sensitive, specific, and efficient diagnostic method for detecting E. platys in clinical samples.

並列關鍵字

Canine infectious cyclic thrombocytopenia ； Ehrlichia platys ； Polymerase chain reaction