羊附红细胞体PCR诊断方法的建立及应用
Development and Preliminary Application of PCR Assay Based on the 16S rRNA Gene for Detection of Eperythrozoon ovis in Sheep
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摘要: 根据羊附红细胞体16S rRNA基因序列设计特异性引物,建立了羊附红细胞体的PCR诊断方法.该方法能扩增羊附红细胞体16S rRNA基因片段,对大肠杆菌、羊无形体、羊源支原体、猪附红体和假单孢菌基因组DNA没有扩增出条带,所能检测羊附红细胞体DNA的最低量为1.82 pg/mL.运用所建立的方法对从重庆市荣昌县采集的110份羊血样进行附红细胞体检测,其中13份羊附红体感染阳性,表明该方法特异性和灵敏度高,可用于急性羊附红细胞体病和临床带菌羊的检测.
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关键词:
- 羊附红细胞体,PCR,检测
Abstract: Based on the sequence data of 16S rRNA gene of Eperythrozoon ovis,a set of E.ovis specific primers were designed.No PCR products were amplified from purified DNA of Escherichia coli,Anaplasma ovis,Mycoplasma ovis,Eperythrozoon suis and Pseudomonas putid.It was estimated that as few as1.82 pg/mL E.ovis genomic DNA was detectable by PCR.Using this method,110 blood samples from Rongchang were successfully detected and 13 samples were positive with E.ovis infection.The results showed the PCR method developed was an effective tool for diagnosing E.ovis infection in sheep with high sensitivity and specificity.It will be useful for detecting the acute eperythrozoonosis and latent infected carrier animals. -
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