温氏附红细胞体病PCR诊断方法的建立及其应用

Establishment and Application of PCR Method for Diagnosing Eperythrozoon wenyonii

为建立特异、敏感、快速的温氏附红细胞体诊断方法,根据温氏附红细胞体16S rRNA基因序列(GenBank登录号为AY946266),设计1对种特异性引物,建立了温氏附红细胞体的PCR诊断方法。特异性试验和敏感性试验结果表明,该方法能特异性地扩增温氏附红细胞体16S rRNA基因序列的特异性片段,检测温氏附红细胞体最低DNA量为0.178 fg。利用该方法对湖北省、江苏省和黑龙江省温氏附红细胞体病感染情况进行了流行病学调查,结果显示阳性感染率分别为10.3%、5.3%、45.8%,说明温氏附红细胞体在湖北省、江苏省和黑龙江省均存在,流行病学调查显示湖北省感染率较高、流行较广。

A PCR was developed to detect Eperythrozoon wenyonii in cattle. The specific 314 bp fragment was amplified from 16S rRNA gene of E. wenyonii. The practical limit of detection showed that it had high sensitivity; an approximate DNA of 0. 178 fg was detected by the PCR system and there were no cross among Mycoplasma hyopneumoniae , Babesia orientalis , B. boris, B. bigemina, B. motasi, Anaplasma marginale ,A. central,A, ovis, Trypanosoma evansi , Theileria hirci , T. annulata and E. wenyonii. The 333 blood samples collected from Hubei,Jiangsu and Heilongjiang Province were examined for the presence of E. wenyonii by PCR. The results showed that average infection rate of E. wenyonii were 10.3%,5. 3% and 45. 8%. This data revealed that bovine eperythrozoonosis was present in the three provinces,and present widely in Hubei Province.