多重实时聚合酶链反应与血清学试验和培养比较诊断人类布鲁氏菌病。

Comparison of multiplex real-time polymerase chain reaction with serological tests and culture for diagnosing human brucellosis.

机构信息

Ankara Yıldırım Beyazıt University, Faculty of Medicine, Department of Clinical Microbiology, Ankara, Turkey.

Erzurum Regional Training and Research Hospital, Department of Pediatric Infectious Disease, Erzurum, Turkey.

出版信息

J Infect Public Health. 2019 May-Jun;12(3):337-342. doi: 10.1016/j.jiph.2018.11.008. Epub 2018 Dec 13.

DOI:10.1016/j.jiph.2018.11.008

PMID:30553722

Abstract

OBJECTIVE

Brucellosis is a zoonotic disease with various clinical presentations and early diagnosis is crucial to avoid severe complications. Due to limitations of conventional diagnostic methods, polymerase chain reaction (PCR) based approaches have gained importance in diagnosis.We aimed to evaluate diagnostic value of multiplex real time-PCR (mRT-PCR) in serum samples collected from brucellosis suspected patients by comparision sensitivity of mRT-PCR with those of conventional diagnostic methods.

MATERIAL AND METHODS

A total of 249 serum samples collected from the suspected brucellosis patients admitted to the hospitals in three different provinces were analyzed by serological tests, culture and mRT-PCR. In laboratories of the participating hospital, serum samples were tested for the Brucella specific antibody by commercial serological kits including standart tube agglutination test (STAT), Coombs' test, and immunocapture test (ICT). Blood culture was performed for 153 of the patients in the participating hospital. All serum samples were analyzed for the presence of Brucella DNA by mRT-PCR.

RESULTS

According to laboratory test results, 215 of the 249 suspected cases having comparible clinical data were identified as brucellosis cases. Of the 215 brucellosis cases, 36 were diagnosed as definitive cases, the remaning 179 patients were presumptive cases. Sensitivity of mRT-PCR in the samples that were positive by ICT, STAT, Coombs' test, and blood culture was 70.2%, 77.3%, 83%, and 97.2%, respectively. By using mRT-PCR, additional 17 suspected patients were diagnosed as presumptive cases. Among the mRT-PCR positive serum samples, Brucella abortus was detected in 3 samples (1.9%), the remaining 156 samples (98.1%) had B. melitensis DNA.

CONCLUSION

Our results indicate that mRT-PCR can be considered a useful diagnostic tool in patients who have negative serologic test results, and in detection of Brucella species.

摘要

目的

布鲁氏菌病是一种人畜共患疾病，临床表现多样，早期诊断对于避免严重并发症至关重要。由于常规诊断方法的局限性，聚合酶链反应（PCR）方法在诊断中得到了重视。本研究旨在通过比较多重实时 PCR（mRT-PCR）与常规诊断方法的敏感性，评估其在疑似布鲁氏菌病患者血清样本中的诊断价值。

材料与方法

分析了来自三个不同省份的疑似布鲁氏菌病患者的 249 份血清样本，这些患者分别在不同的医院接受了血清学检测、培养和 mRT-PCR 检测。在参与医院的实验室中，使用商业血清学试剂盒（包括标准试管凝集试验（STAT）、库姆斯试验和免疫捕获试验（ICT））检测血清中布鲁氏菌特异性抗体。对 153 名参与医院的患者进行了血培养。所有血清样本均通过 mRT-PCR 分析是否存在布鲁氏菌 DNA。

结果

根据实验室检测结果，对 249 例具有可比临床数据的疑似病例进行了分析，其中 215 例被诊断为布鲁氏菌病。在这 215 例布鲁氏菌病中，36 例为确诊病例，其余 179 例为疑似病例。在 ICT、STAT、库姆斯试验和血培养阳性的样本中，mRT-PCR 的敏感性分别为 70.2%、77.3%、83%和 97.2%。通过使用 mRT-PCR，另外 17 例疑似患者被诊断为疑似病例。在 mRT-PCR 阳性的血清样本中，检测到 3 份（1.9%）布鲁氏菌流产亚种，其余 156 份（98.1%）样本含有布鲁氏菌 melitensis DNA。