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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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