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首页> 外文期刊>Iranian Journal of Basic Medical Sciences >DETECTION AND DISCRIMINATION OF TWO BRUCELLA SPECIES BY MULTIPLEX REAL?TIME PCR AND HIGH?RESOLUTION MELT ANALYSIS CURVE FROM HUMAN BLOOD AND COMPARISON OF RESULTS USING RFLP
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DETECTION AND DISCRIMINATION OF TWO BRUCELLA SPECIES BY MULTIPLEX REAL?TIME PCR AND HIGH?RESOLUTION MELT ANALYSIS CURVE FROM HUMAN BLOOD AND COMPARISON OF RESULTS USING RFLP

机译：多重实时荧光定量PCR和高分辨熔体分析曲线从人类血液中检测和鉴定两种布鲁氏菌，并利用RFLP比较结果

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Objective(s): Rapid and accurate detection of Brucella abortus and Brucella melitensis from clinical samples is so important because antibiotic treatment has major side effects. This study reveals a new method in detection of clinical samples of brucellosis using real ? time PCR and high ? resolution melt (HRM) curve analysis.Materials and Methods: 160 brucellosis suspicious samples with more than 1/80 serum antibody titers were collected and the results were compared with the RFLP method. In order to amplify the sequences for HRM analysis, vdcc, int ? hyp and glk and for RFLP, omp2a and omp2b with PstI and Hinf1 restriction endonuclease were used. At last, the accuracy and specificity of the two methods were compared with each other.Results: Out of these 160 samples, multiplex real time PCR showed 108 positive samples (67.5%), including 56%B. melitensis and 44% B. abortus; whereas in PCR ? RFLP 52 out of 160 samples were positive, where recognition of two species were accordant with HRM analysis, separation was based on the size of the amplified fragment. Using the designed primers and performing the assay, we confirmed this method to be much faster and have lower cost with more than 99% accuracy compared to methods such as RFLP.Conclusion: The present study showed that this technique, which scans gene segments and creates an analysis pattern for detection of clinical samples, is useful and more dominant compared with PCR ? RFLP.Thus, this method can be used for brucellosis detection, and clinical and epidemiological research.
机译：目的：快速准确地从临床样本中检测流产布鲁氏菌和布鲁氏菌非常重要，因为抗生素治疗具有重大的副作用。这项研究揭示了一种使用真正的？检测布鲁氏菌病临床标本的新方法。 PCR时间高吗？材料与方法：收集160份血清抗体滴度超过1/80的布鲁氏菌病可疑样品，并将结果与​​RFLP方法进行比较。为了扩增用于HRM分析的序列，请使用vdcc，int？ hyp和glk，对于RFLP，使用带有PstI和Hinf1限制性核酸内切酶的omp2a和omp2b。最后，对两种方法的准确性和特异性进行了比较。结果：在这160份样品中，多重实时PCR显示108份阳性样品（67.5％），其中56％为B。 melitensis和44％流产的B.而在PCR中呢？ 160个样本中的RFLP 52呈阳性，其中两种物质的识别与HRM分析相符，分离基于扩增片段的大小。通过使用设计的引物并进行测定，我们证实了该方法与RFLP等方法相比，速度更快且成本更低，准确率高达99％。结论：本研究表明，该技术可扫描基因片段并产生一种用于检测临床样品的分析模式，与PCR相比是否有用且占主导地位？ RFLP。因此，该方法可用于布鲁氏菌病的检测以及临床和流行病学研究。

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