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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2009, Vol. 3 Issue (1) : 100-103     DOI: 10.1007/s11703-009-0014-1
RESEARCH ARTICLE |
Detection of Eperythrozoon wenyoni by PCR assay
Jian WANG1, Yutao ZHU1, Jianhua QIN1(), Jian WANG2, Fumei ZHANG2, Yuelan ZHAO3
1. College of Animal Science and Technology, Agricultural University of Hebei, Baoding 071001, China; 2. Department of Food Science, Hebei North University, Zhangjiakou 075000, China; 3. College of Traditional Chinese Veterinary Science, Agricultural University of Hebei, Dingzhou 073000, China
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Abstract  

The objective of this research was to develop a detection method for Eperythrozoon wenyoni infection using polymerase chain reaction (PCR) assay technique. A pair of primers was designed and synthesized according to the conservative sequence 16S rRNA. The PCR assay was performed with the primers. A 985-bp fragment was amplified by using PCR. The amplified fragments with the expected size were identified by EcoR I restriction digestion. The crossing-reaction, specific-reaction and duplicate-reaction indicated that the PCR method is a specific, sensitive, fast and effective method for diagnosing E. wenyoni infection at group level.

Keywords cattle      Eperythrozoon wenyoni      PCR     
Corresponding Authors: QIN Jianhua,Email:qjhqqq@126.com   
Issue Date: 05 March 2009
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-009-0014-1     OR     http://academic.hep.com.cn/fag/EN/Y2009/V3/I1/100
Fig.1  Purified genomic DNA of
Note: M stands for DL2000 DNA marker; 1 stands for purified DNA of .
Fig.2  DNA fragment amplified by PCR
Note: M stands for DL2000 DNA marker, 1 stands for PCR products, and 2 stands for the negative control.
Fig.3  Enzyme-cutting identification of amplified products
Note: M stands for DL2000 DNA marker, and l stands for R I digestion results.
Fig.4  Sensitivity of PCR products
Note: M stands for DL2000 DNA marker; 1-5 for the PCR results with the serial concentration of template DNA of 1 ng, 100 pg, 10 pg, 1 pg, and 0.1 pg, respectively.
Fig.5  The result of specificity of PCR
Note: M stands for DL2000 DNA marker; 1, 2, 3, and 4 stand for the amplified fragment of , the amplified fragment of , the amplified fragment of , and the amplified fragment of , respectively.
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