Detection of Brucella melitensis in semen using the polymerase chain reaction assay
Detection of Brucella melitensis in semen using the polymerase chain reaction assay
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DOI：
10.1016/s0378-1135(01)00401-1
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发表时间：
2001-10-22
影响因子：
3.3
通讯作者：
Ibrahim, AK
中科院分区：
文献类型：
--
作者：
Amin, AS;Hamdy, MER;Ibrahim, AK
对聚合酶链反应（PCR）检测牛和绵羊精液中羊种布鲁氏菌DNA的方法进行了评价。由于精液中含有不同的抑制PCR扩增的成分，因此在进行靶向DNA扩增之前，使用一种方案从牛和绵羊精液样品中纯化布鲁氏菌DNA。用PCR方法分析布鲁氏菌自然污染精液的分离组分时，大多数B。精液和非精液成分中均存在羊种DNA，PCR检测结果为B.方法：将不同精液组分中羊种布氏菌DNA的培养结果与传统精液布氏菌培养结果进行比较。PCR法比传统的培养法敏感，在120份精液中检出12份（10%）布鲁氏菌DNA，而直接培养法仅检出7份（5.8%）。PCR检测限为100 CFU/ml精液。此外，聚合酶链反应的结果在一天内就可以得到，而布鲁氏菌的分离和鉴定需要几天甚至几周的时间。PCR可作为检测B的补充试验。在精液中的melitensis。(C)2001爱思唯尔科技有限公司。保留所有权利。
An evaluation of the polymerase chain reaction (PCR) for detection of Brucella melitensis DNA in bovine and ovine semen was performed. Since semen contains different components that inhibit PCR amplification, a protocol was used to purify Brucella-DNA from bovine and ovine semen samples prior to conducting amplification of the targeted DNA. When separated fractions of naturally Brucella contaminated semen were analyzed by the PCR, most of B. melitensis DNA were present in the seminal fluid and non-sperm fractions.The PCR examination results for detection of B. melitensis DNA in different semen fractions were compared with the results for traditional cultural methods of Brucella from semen. The PCR was more sensitive than the traditional cultural methods since it detected Brucella-DNA in 12 (10%) out of 120 semen samples while direct culture detected only 7 (5.8%) in the same semen samples. The limit of detection by PCR was 100 CFU/ml of semen. In addition, the results of PCR were available in one day, whereas isolation and identification of Brucella organisms required days or even weeks. The PCR may be used as a supplementary test for detection of B. melitensis in semen. (C) 2001 Elsevier Science B.V. All rights reserved.