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| Establishment and Preliminary Application of a Duplex PCR Assay for Detecting Brucella and Coxiella burnetii |
| PENG Wu-li1,2, LUO Zhan-rong3, JI Xin-cheng2, YUN Li-juan2, SHI Qian2, YU Xue-hui2 |
1. College of Life Science of Shihezi University, Shihezi 832000, China;
2. Xinjiang Exit-import Inspection and Quarantine Bureau, Urumqi 830063, China;
3. 69016 Armed Forces of Chinese People's Liberation Army, Urumqi 830001, China |
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Abstract To establish a duplex PCR assay for simultaneously detecting Brucella and Coxiella burnetii, two genus-specific genes sequences according to the Bp26 of Brucella spp. and IS1111a of Coxiella burnetii were obtained in GenBank and two specific primers pairs were designed respectively. After optimizing the PCR reactions and amplification conditions, the duplex assay was finally established. This assay had better specificity, repeatability and higher sensitivity. Using this assay to detect 172 samples including bloods,serums, fetuses and milk of abortion cattle in clinic,53 brucellosis positive samples and 10 Q fever positive samples were detected out, the positive rates were separately 30.8% and 5.8%, 2 co-infection positive samples were detected out also and the positive rate was 1.2%. The above preliminary clinical results indicated this assay can be used for safely, simultaneously, rapidly and sensibly detecting Brucella and Coxiella burnetii.
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Received: 24 October 2013
Published: 25 February 2014
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[1] Seleem M N, Boyle S M, Sriranganathan N. Brucellosis: a re-emerging zoonosis. Vet Microbiol, 2010, 140(3): 392-398.
[2] Hartzell J D, Wood-Morris R N, Martinez L J, et al. Q fever: epidemiology, diagnosis, and treatment. Mayo Clin Proc, 2008, 83(5): 574-579.
[3] Seshadri R, Paulsen I T, Eisen J A, et al. Complete genome sequence of the Q-fever pathogen Coxiella burnetii. Proc Natl Acad Sci, 2003, 100(9): 5455-5460.
[4] 俞树荣. 中国 Q 热研究进展. 中华流行病学杂志, 2000, 21(6): 456-459. Yu S R. Proceeding on Q fever in China. Chinese Journal of Epidemiology, 2000, 21(6): 456-459.
[5] Wang S M, Liu C C. Enterovirus 71: epidemiology, pathogenesis and management. Expert Rev Anti Infect Ther, 2009, 7(6): 735-742.
[6] Schelling E, Diguimbaye C, Daoud S, et al. Brucellosis and Q-fever seroprevalences of nomadic pastoralists and their livestock in Chad. Prev Vet Med, 2003, 61(4): 279-293.
[7] Mirnejad R, Doust R H, Kachuei R, et al. Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR. Asian Pac J Trop Med, 2012, 5(1): 24-28.
[8] Wang X L, Xiong X L, Graves S, et al. Protein array of Coxiella burnetii probed with Q fever sera. Sci China Life Sci, 2013, 56(5): 453-459.
[9] Gupta V K, Verma D K, Rout PK, et al. Polymerase chain reaction (PCR) for detection of Brucella melitensis in goat milk. Small Ruminant Res, 2006, 65(1): 79-84.
[10] Kersh G J, Fitzpatrick K A, Self J S, et al. Presence and persistence of Coxiella burnetii in the environments of goat farms associated with a Q fever outbreak. Appl Environ Microbiol, 2013, 79(5): 1697-1703.
[11] Zhang L, Fu X, Zhang J. Q fever endocarditis with multi-organ complication: a case report. Chin Med J (Engl), 2006, 119(18): 1580.
[12] Karakousis PC, Trucksis M, Dumler J S. Chronic Q fever in the United States. J Clin Microbiol, 2006, 44(6):2283-2287.
[13] 程仕虎, 刘又宁, 李朝霞, 等. 44 例被误诊为普通性肺炎的 Q 热柯克斯体肺炎病人的回顾性分析. 科学技术与工程, 2007, 10(7): 2335-2337. Cheng S H, Liu Y N, Li C X, et al. Looking back analysis for Q fever Coxiella Burnetu which be misdiagnosis common pneumonia of 44 examples. Science Technology and Engineering, 2007, 10(7): 2335-2337.
[14] 朱晓宇, 魏建春, 张慧娟, 等. 一次不明原因发热疫情中病人血清的 Q 热抗体检测. 中国卫生检验杂志, 2008, 18(6): 1130-1131. Zhu X Y, Wei J C, Zhang H J, et al. Detection of antibody against Q fever from sera provided by patients in an epidemic of fever of unknown origin. Chinese Journal of Health Laboratory Technology, 2008, 18(6): 1130-1131. |
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