Sifuentes-Rincón A M, Revol A, Barrera-Saldaña H A
Departamento de Bioquímica, Facultad de Medicina, Universidad Autónoma de Nuevo León, Monterrey, México.
Mol Med. 1997 Nov;3(11):734-9.
Brucelosis is a severe acute febrile disease caused by bacteria of the genus Brucella. Its current diagnosis is based on clinical observations that may be complemented by serology and microbiological culture tests; however, the former is limited in sensitivity and specificity, the latter is time consuming. To improve brucelosis diagnosis we developed a test which is specific and sensitive and is capable of differentiating the six species of Brucella.
Four primers were designed from B. abortus sequences at the well-conserved Omp2 locus that are able to amplify the DNAs of all six species of Brucella.
Our test detected all six species of Brucella. Their differentiation resulted directly from differences in the amplification patterns or was achieved indirectly using a RFLP present in one of the PCR products. The sensitivity and specificity of the new test were then determined; it was applied successfully in confirming the diagnosis of a patient whose clinical history and serology indicated infection with Brucella.
The results make possible the use of a PCR test for Brucella detection and differentiation without relying on the measurement of the antibodies or microorganism culture. Our first results showed that the PCR test can confirm the presence of Brucella in blood samples of infected patients.
布鲁氏菌病是由布鲁氏菌属细菌引起的一种严重的急性发热性疾病。其目前的诊断基于临床观察，血清学和微生物培养测试可作为补充；然而，前者在敏感性和特异性方面存在局限性，后者则耗时较长。为了改进布鲁氏菌病的诊断，我们开发了一种具有特异性和敏感性且能够区分六种布鲁氏菌的检测方法。
根据流产布鲁氏菌在高度保守的Omp2位点的序列设计了四种引物，这些引物能够扩增所有六种布鲁氏菌的DNA。
我们的检测方法检测出了所有六种布鲁氏菌。它们的区分直接源于扩增模式的差异，或者通过使用其中一种PCR产物中存在的限制性片段长度多态性（RFLP）间接实现。然后确定了新检测方法的敏感性和特异性；它成功应用于确诊一名临床病史和血清学表明感染布鲁氏菌的患者。
这些结果使得无需依赖抗体检测或微生物培养即可使用PCR检测方法来检测和区分布鲁氏菌成为可能。我们的初步结果表明，PCR检测方法能够确认感染患者血液样本中存在布鲁氏菌。
