Background

This study aims to evaluate the efficiency of the TaqMan real-time PCR and serological methods in detecting Brucella spp. in clinical specimens that have been collected from suspected patients in Ardabil, Iran.

Methods

In this cross-sectional study, a total of 113 consecutive patients suspected of brucellosis who were referred to the three hospitals in Ardabil province were selected. In the first step, the diagnosis of brucellosis was performed by serological methods including the Rose Bengal slide agglutination test, Wright test, 2-ME test, and BrucellaCapt test. In the next step, TaqMan real-time PCR with primer and probe targeting the bcsp31 gene was used for the detection of Brucella spp. Specificity, sensitivity, and positive and negative predictive values of the TaqMan real-time PCR assay were calculated.

Results

Among 113 suspected patients with different clinical manifestations, the Rose Bengal slide agglutination test, Wright test, and 2-ME test were positive in 60 cases; however, the BrucellaCapt test titer was 1:160 for one patient. Six patients had high initial serum antibody titers; 2-ME titers of ≥1:640; STA titers of ≥1:1280; BrucellaCapt titers of ≥ 1:2560. Among positive cases, no correlation was observed among gender, age, and life (residence) in urban or rural areas. The TaqMan real-time PCR was positive in 35% of all 60 positive cases. The comparison of the results of the BrucellaCapt and TaqMan real-time PCR methods revealed that 19 out of 54 (35.2%) and 2 out of 6 (33.4%) BrucellaCapt positive cases with titers of >1:320 and ≤ 1:320 were positive, respectively. The sensitivities and specificities of the TaqMan real-time PCR assay were 49.1% and 100% respectively.

Conclusion

The sensitivity of the TaqMan real-time PCR assay was low in the diagnosis of brucellosis, while the BrucellaCapt test turned out to be a very valuable, sensitive, and specific test for the diagnosis of brucellosis in suspected patients and, thus, can provide reliable results in medical laboratories.

中文翻译：

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评估TaqMan实时PCR和血清学方法在布鲁氏菌属检测中的效率。从伊朗阿尔达比勒（Aldabil）的可疑患者那里收集的临床标本中。

背景

这项研究旨在评估TaqMan实时PCR和血清学方法检测布鲁氏菌属种的效率。从伊朗阿尔达比勒（Aldabil）的可疑患者那里收集的临床标本中。

方法

在这项横断面研究中，选择了总共113例疑似布鲁氏菌病的患者，这些患者被转诊至Ardabil省的三家医院。第一步，通过血清学方法对布鲁氏菌病进行诊断，包括Rose Bengal玻片凝集试验，Wright试验，2-ME试验和BrucellaCapt试验。下一步，将TaqMan实时PCR的引物和靶向bcsp31基因的探针用于布鲁氏菌属的检测。计算TaqMan实时PCR分析的特异性，敏感性以及阳性和阴性预测值。

结果

在113例不同临床表现的可疑患者中，玫瑰孟加拉玻片凝集试验，莱特氏试验和2-ME试验阳性60例。但是，一名患者的BrucellaCapt检测效价为1：160。六例患者的初始血清抗体滴度很高。2-ME滴度≥1：640; STA滴度≥1：1280；BrucellaCapt效价≥1：2560。在阳性病例中，未发现城市或农村地区的性别，年龄和生活（居住）之间存在相关性。在所有60例阳性病例中，TaqMan实时PCR阳性35％。BrucellaCapt和TaqMan实时PCR方法结果的比较显示，滴度> 1：320和≤1：320的BrucellaCapt阳性病例中54例中的19例（35.2％）和6例中的2例（33.4％）为正面分别。TaqMan实时PCR分析的灵敏度和特异性为49。

结论

TaqMan实时PCR测定法在布鲁氏菌病的诊断中灵敏度较低，而BrucellaCapt检测结果对于可疑患者的布鲁氏菌病诊断是非常有价值，灵敏且特异的检测，因此可以提供可靠的导致医学实验室。