采用组合 PCR 技术同时检测和区分流产布鲁氏菌和马耳他布鲁氏菌。

Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR.

机构信息

Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

出版信息

Asian Pac J Trop Med. 2012 Jan;5(1):24-8. doi: 10.1016/S1995-7645(11)60239-3.

DOI:10.1016/S1995-7645(11)60239-3

PMID:22182638

Abstract

OBJECTIVE

To evaluate simultaneous detection and differentiates of Brucella abortus (B. abortus) and Brucella melitensis (B. melitensis) through the combinatorial PCR method.

METHODS

This study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals. Identification and differentiation of each species using the size of the PCR product were determined. To determine the specificity of the method, bacteria close to the genus Brucella were used. Finally, to confirm PCR products, In addition to the products sequence, RFLP was performed on PCR products using restriction enzymes.

RESULTS

The method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B. abortus and B. melitensis with high specificity and sensitivity in clinical samples. Differentiation of species is based on the resulting bands; therefore, the band 494 bp for B. abortus and 733 bp for B. melitensis were obtained. RFLP and sequencing results confirmed PCR results.

CONCLUSIONS

The results of this study shows that without routine diagnostic methods such as culture and serology tests, using the molecular method of combinatorial PCR, important species of Brucella can be simultaneously identified and differentiated in clinical samples.

摘要

目的

通过组合 PCR 方法评估布鲁氏菌属（Brucella abortus，B. abortus）和流产布鲁氏菌（Brucella melitensis，B. melitensis）的同时检测和区分。

方法

本研究设计了三组引物，可同时鉴定和区分人类和动物中两种主要的致病性布鲁氏菌。通过 PCR 产物的大小确定每种物种的鉴定和区分。为了确定该方法的特异性，使用了与布鲁氏菌属密切相关的细菌。最后，除了对 PCR 产物进行测序外，还使用限制性内切酶对 PCR 产物进行 RFLP 分析。

结果

本研究中优化的组合 PCR 方法可在临床样本中同时检测和区分 B. abortus 和 B. melitensis，具有高度特异性和敏感性。通过产生的条带来区分物种，因此得到 494 bp 的条带为 B. abortus，733 bp 的条带为 B. melitensis。RFLP 和测序结果证实了 PCR 结果。