Comparison of nested PCR with immunofluorescent-antibody assay for detection of Ehrlichia canis infection in dogs treated with doxycycline
Comparison of nested PCR with immunofluorescent-antibody assay for detection of Ehrlichia canis infection in dogs treated with doxycycline
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DOI：
10.1128/jcm.35.7.1852-1855.1997
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发表时间：
1997-07-01
影响因子：
9.4
通讯作者：
Bartsch, R
中科院分区：
文献类型：
--
作者：
Wen, BH;Rikihisa, Y;Bartsch, R
用套式聚合酶链式反应从犬埃立克体感染细胞中扩增出部分16S rRNA基因，该方法具有较高的特异性，不扩增与查菲埃立克体、小鼠埃立克体、蠕虫新立克次体和SF毒剂密切相关的16S rRNA基因。该方法与Southern杂交一样灵敏，只检测到0.2pg的犬E.canis DNA。用这种方法，4只实验感染犬的血样在血清转换前或血清转换时的第4天全部阳性，来自亚利桑那州和德克萨斯州的105只狗的血样全部阳性。用套式聚合酶链式反应和免疫荧光抗体(IFA)试验对俄亥俄州犬非流行区的30份血样进行了检测。来自亚利桑那州和得克萨斯州的大约84%的狗在提交血液样本之前接受过多西环素治疗，在亚利桑那州和得克萨斯州的样本中，46个样本为PCR阳性(44%)，80个样本为IFA阳性(76%)。80份IFA阳性标本中有43份阳性(54%)，25份IFA阴性标本中有22份(88%)为阴性，俄亥俄州无一份IFA阳性，5份阳性(17%)。结果表明，套式PCR检测犬艾美耳球虫具有很高的敏感性和特异性，尤其是在犬艾美耳球虫流行地区，比IFA更有助于评估抗生素治疗后细菌的清除情况。
A partial 16S rRNA gene was amplified in Ehrlichia canis-infected cells by nested PCR, The assay was specific and did not amplify the closely related Ehrlichia chaffeensis, Ehrlichia muris, Neorickettsia helminthoeca, and SF agent 16S rRNA genes, The assay was as sensitive as Southern hybridization, detecting as little as 0.2 pg of E. canis DNA, By this method, all blood samples from four dogs experimentally infected with E. canis were positive as early as day 4 postinoculation, which was before or at the time of seroconversion, One hundred five blood samples from dogs from Arizona and Texas (areas off. canis endemicity) and 30 blood samples from dogs from Ohio (area of E. canis nonendemicity) were examined by nested PCR and immunofluorescent-antibody (IFA) test. Approximately 84% of dogs from Arizona and Texas had been treated with doxycycline before submission of blood specimens, Among Arizona and Texas specimens, 46 samples were PCR positive (44%) and 80 were IFA positive (76%). Forty-three of 80 IFA-positive samples (54%) were PCR positive, and 22 of 25 IFA-negative samples (88%) mere negative in the nested PCR, None of the Ohio specimens were IFA positive, but 5 specimens were PCR positive (17%), Our results indicate that the nested PCR is highly sensitive and specific for detection of E. canis and may be more useful in assessing the clearance of the organisms after antibiotic therapy than IFA, especially in areas in which E. canis is endemic.