Mohseni Khashayar, Mirnejad Reza, Piranfar Vahab, Mirkalantari Shiva
Student Research Committee, Semnan University of Medical Sciences, Semnan, Iran.
Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Iran J Pathol. 2017 Fall;12(4):371-376. Epub 2017 Oct 1.
BACKGROUND & OBJECTIVE: Since the symptoms of Brucellosis are often atypical and nonspecific, using clinical signs alone to diagnose brucellosis is not advised; therefore, the diagnosis relies predominantly on laboratory testing. Currently, molecular, serological, and microbiological methods are used for diagnosis of this disease. In this study we examined ELISA, PCR and serum agglutination (SAT) methods on human patient serum samples.
A total of 100 serum samples were collected from suspected patients. Fifty serum samples gave a positive result with the Wright test. The ELISA method was first employed on all samples for the detection of IgG and IgM antibodies against . Subsequently, the rapid PCR methodology was used to identify presence of genome in 500 µL of each serum sample. The B4/B5 primer pair was used for PCR amplification.
Out of the 100 serum samples obtained from patients with suspected brucellosis, 50 samples tested positive by SAT and displayed high titers of 1/160. Of these 50 positive samples, 49 samples were positive as per the ELISA test whereas one sample tested negative. The PCR test was conducted on all 100 serum samples and results showed that the 45 serum samples that gave a positive agglutination test were also positive by PCR.
Various laboratory methods have been used or introduced for the detection of . Molecular methods such as PCR, a rapid and sensitive method for detection of bacteria, have also been reported. Based on the results of this study, we propose that the simultaneous use of serology and molecular techniques has the potential to overcome limitations of detection thereby enabling the selection of appropriate treatment for the patient.
由于布鲁氏菌病的症状通常不典型且无特异性，不建议仅依靠临床症状来诊断布鲁氏菌病；因此，诊断主要依赖实验室检测。目前，分子、血清学和微生物学方法都用于该疾病的诊断。在本研究中，我们对人类患者血清样本检测了酶联免疫吸附测定（ELISA）、聚合酶链反应（PCR）和血清凝集试验（SAT）方法。
从疑似患者中总共收集了100份血清样本。50份血清样本的wright试验结果呈阳性。首先对所有样本采用ELISA方法检测抗……的IgG和IgM抗体。随后，使用快速PCR方法鉴定每份500微升血清样本中……基因组的存在情况。B4/B5引物对用于PCR扩增。
在从疑似布鲁氏菌病患者获得的100份血清样本中，50份样本经SAT检测呈阳性，且滴度高达1/160。在这50份阳性样本中，49份样本经ELISA检测呈阳性，而1份样本检测呈阴性。对所有100份血清样本进行了PCR检测，结果显示，45份凝集试验呈阳性的血清样本经PCR检测也呈阳性。
已经使用或引入了各种实验室方法来检测……。也有报道称分子方法如PCR是一种快速且灵敏的细菌检测方法。基于本研究结果，我们建议同时使用血清学和分子技术有可能克服检测的局限性，从而能够为患者选择合适的治疗方法。
