Please wait a minute...
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
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
Download: PDF(123 KB)   HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks

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,   
Issue Date: 05 March 2009
URL:     OR
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.
1 Adler S, Ellenbogen V (1934). A note on two new blood parasites of cattle, Eperythrozoon and Bartonella. Journal of Comparative Pathology , 47: 220-221
2 Baljer G, Heinritzi, Wieler L (1989). Indirect hemagglutination for Eperythrozoon suis detection in experimentally and spontaneously infected swine. Zentralbl Veterinarmed B , 36(6): 417-423
3 Cha H B, Liu S G (2002). The study of current situation and prospect of eperythrozoonosis in pigs. Chinese Journal of Animal Husbandry and Veterinary Medicine , 18(9): 6-9 (in Chinese)
4 Chen M (2006). Development of a PCR diagnostic assay for E. wenyoni infection and its pathogeny primary biology study. Dissertation for the Doctoral Degree . Nanning: Guangxi University (in Chinese)
5 Daddow K N (1977). A complement fixation test for the detection of Eperthrozoon infection in sheep. Aust Vet Res , 53(3): 139-143
doi: 10.1111/j.1751-0813.1977.tb00140.x
6 Hsu F S, Liu M C, Chou S M, Zachary J F, Smith A R (1992). Evaluation of an enzyme-linked immunosorbent assay for detection of Eperythrozoon suis antibodies in swine. Am J Vet Res , 53(3): 352-354
7 Hua S (1970). The detection of eperythrozoonosis in pigs by IFA. Chinese Journal of Veterinary Science and Technology , 8: 57-59 (in Chinese)
8 Huang X, Qin Y F, Mo L C, Hu J, Lu W J, Huang S B, Zhao G M (2003). Diagnosis and prevention on bovine Eperythrozoonosis. Guangxi Journal of Animal Husbandry and Veterinary Medicine , 19(2): 89-91 (in Chinese)
9 Lang F M, Ferrier G R, Nicholls T J (1987). Detection of antibodies to Eperythrozoon ovis by the use of an enzyme-linked immunosorbent assay. Res Vet Sci , 43(2): 249-252
10 Lin J (2000). Eperythrozoonosis. Journal of Medical Postgraduates , 13(1): 59-62 (in Chinese)
11 Mason R W, Statham P (1991). The determination of the level of Eperythrozoon ovis parasitaemia in chronically infected sheep and its significance to the spread of infection. Aust Vet J , 68(3): 115-116
doi: 10.1111/j.1751-0813.1991.tb00771.x
12 Nicholls T J, Veal P I (1986). The prevalence of Eperthrozoon ovis infection in swine weaner and adult sheep in north eastern. Victoria Aust Vet , 63(4): 118-120
doi: 10.1111/j.1751-0813.1986.tb07678.x
13 Sambrook J, Russell D W (2001). Molecular Cloning: a Laboratory Manual. (in Chinese, trans. Huang Pei-tang). 3rd ed. Beijing: Science Press, 611-614
14 Schuller W, Heinritzi K, al-Nuktha S, K?lbl S, Schuh M (1990). Serologic progression studies using CF and ELISA for the detection of antibodies against Eperythrozoon suis infection of swine. Berl Munch Tieraerztl Wochenschr , 103: 9-12
15 Shang D Q (1994). The study of current situation and prospect of eperythrozoonosis. Chinese Journal of Epidemiology , 15(4): 234-240 (in Chinese)
16 Smith A R, Rahn T (1975). An indirect hemagglutination test for the diagnosis of Eperythrozoon suis infection in swine. Am J Vet Res , 36(9): 1319-1321
17 Splitter E J (1985). The complement-fixation test in diagnosis of Eperythrozoonsis in swine. JAVMA , 132: 47-49
18 Vandervoort J M, Bourne C, Carson R L (2001). Use of a polymerase chain reaction assay to detect infection with Eperythrozoon wenyoni in cattle. J Am Vet Med Assoc , 219(10): 1432-1434
doi: 10.2460/javma.2001.219.1432
19 Zhang H J, Xing M Q, Zhang J F, Qin Z H, Gu W J, Cai J P (2005). Sequencing and phylogenetic analysis of the 16S rRNA gene of Eperythrozoon suis. Chinese Journal of Animal and Veterinary Sciences , 36(6): 596-601 (in Chinese)
20 Zhang S F, Zhang G H (2006). Development of a Dot-ELISA method for detection of Eperythrozoon suis. Chinese Journal of Preventive Veterinary Medicine , 28(1): 96-97 (in Chinese)
21 Zinn G M, Jesse G X, Dobson A X (1952). Effect of eperythrozoonosis on sow productivity. JAVMA , 182(4): 369-371
[1] Daling FENG, Shuxin XUAN, Aixia GU, Airu MA, Jiuhuan LI, Shuxing SHEN. PCR-based screening of BAC clones of different chromosomes in Chinese cabbage[J]. Front Agric Chin, 2011, 5(4): 524-528.
[2] Zhongbo XIA, Jihong XING, Xuan WANG, Bin ZHAO, Jianmin HAN, Jingao DONG. Screening of conidium development mutant of Botrytis cinerea and functional analysis of the related gene[J]. Front Agric Chin, 2011, 5(4): 479-485.
[3] Qian WANG, Wei ZHAO, Zhimin HAO, Jingao DONG. mRNA level of PKA-c gene in Setosphaeria turcica with different nutrition sources under metal ion or osmotic stress[J]. Front Agric Chin, 2011, 5(3): 361-365.
[4] John Yew Huat TANG, Farinazleen Mohamad GHAZALI, Abdul Aziz SALEHA,, Yoshitsugu NAKAGUCHI, Mitsuaki NISHIBUCHI, Son RADU. MPN-PCR enumeration of Campylobacter spp. in raw chicken meats and by-products[J]. Front Agric Chin, 2010, 4(4): 501-506.
[5] Hongmiao SHEN, Dangyue HAN, Fenghuan YANG, Qiuxian HUANG, Longxian RAN, Zhengnan LI, . Detection of indigenous endophytic bacteria in Eucalyptus urophylla in vitro conditions[J]. Front. Agric. China, 2010, 4(1): 37-41.
[6] Qiaoyun WENG, Jihong XING, Jingao DONG, Zhiyong LI, Zhiping DONG, . Expression analysis of RUS1 and construction of RUS1 plant expressing vector[J]. Front. Agric. China, 2010, 4(1): 31-36.
[7] Tinghui LIU, Wei GUO, Weiming SUN, Yongxiang SUN. Biological characteristics of Bacillus thuringiensis strain Bt11 and identification of its cry-type genes[J]. Front Agric Chin, 2009, 3(2): 159-163.
[8] Yan ZHANG, Wenxiang YANG, Yaning LI, Daqun LIU, Ting ZHANG. A multiplex PCR method for detection of Clavibacter michiganensis subsp. michiganensis with co-amplification of its host DNA[J]. Front Agric Chin, 2009, 3(2): 140-145.
[9] Haiqiang LU, Hongwei YU, Runfang GUO, Yingmin JIA. Improvement of megaprimer method for site-directed mutagenesis and its application to phytase[J]. Front Agric Chin, 2009, 3(1): 43-46.
[10] ZHANG Xiaohui, XU Shangzhong, GAO Xue, REN Hongyan, CHEN Jinbao. Cloning and bioinformatics analysis of cDNA encoding cattle 4 gene[J]. Front. Agric. China, 2008, 2(4): 493-497.
[11] YE Manhong, CAO Honghe, LI Hongbin, CHEN Jilan, ZHAO Guiping, ZHENG Maiqing. Association of polymorphisms in adipocyte fatty acid binding protein gene with fat-related traits in chicken[J]. Front. Agric. China, 2008, 2(4): 474-479.
[12] WANG Qiong, YANG Chaowu, LIU Yiping, QIU Mohan, ZHU Qing, JIANG Xiaosong, DU Huarui. Correlation analysis of relationships between polymorphisms of high quality chicken gene and slaughter and meat quality traits[J]. Front. Agric. China, 2008, 2(4): 512-518.
[13] ZHANG Xiaohui, XU Shangzhong, GAO Xue, ZHANG Lupei, REN Hongyan, CHEN Jinbao. The application of asymmetric PCR-SSCP in gene mutation detecting[J]. Front. Agric. China, 2008, 2(3): 361-364.
[14] LI Xianglong, ZHANG Zengli, JIA Qing, WANG Lize, GONG Yuanfang, LIU Zhengzhu. mtDNA D-loop of Chinese main indigenous sheep breeds using PCR-RFLP[J]. Front. Agric. China, 2007, 1(3): 352-356.
[15] SUN Feifei, HOU Xilin, LI Ying, CUI Xiumin. Molecular cloning and characterization of nitrate reductase gene cDNA from non-heading Chinese cabbage[J]. Front. Agric. China, 2007, 1(2): 188-192.
Full text