Han Xiangan, Ding Chan, He Liang, Hu Qinghai, Yu Shengqing
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 518 Ziyue Road, Shanghai 200241, People's Republic of China.
Avian Dis. 2011 Sep;55(3):379-83. doi: 10.1637/9602-112610-Reg.1.
Riemerella anatipestifer (RA) infections cause major economic losses in the duck industry. Detection of RA using conventional assays is time-consuming and laborious. In this study, a simple and rapid assay for the detection of RA was established based on the GroEL gene sequence of RA using loop-mediated isothermal amplification (LAMP) with a set of six primers (two outer primers, two inner primers, and two loop primers). This assay was able to detect all the tested RA strains with different serotypes. A minimum of 10 colony-forming units (CFU) of RA was detected, which represents 50-fold higher sensitivity than that of the standard polymerase chain reaction (PCR) method. This assay showed good specificity to RA strains and did not react with any other species of bacteria. The assay is rapidly completed and the amplification is achieved at a minimum of 20 min at 65 C. Furthermore, the assay successfully detected RA in the liver samples of ducklings infected with RA, suggesting that the assay could be used for the clinical diagnosis of RA infection.
鸭疫里默氏菌（RA）感染给养鸭业造成重大经济损失。使用传统检测方法检测RA既耗时又费力。在本研究中，基于RA的GroEL基因序列，利用环介导等温扩增技术（LAMP）和一组六种引物（两条外引物、两条内引物和两条环引物），建立了一种简单快速的RA检测方法。该检测方法能够检测所有测试的不同血清型的RA菌株。检测到的RA最低菌落形成单位（CFU）为10个，其灵敏度比标准聚合酶链反应（PCR）方法高50倍。该检测方法对RA菌株具有良好的特异性，不与任何其他细菌种类发生反应。该检测方法快速完成，在65℃下至少20分钟即可完成扩增。此外，该检测方法成功检测到感染RA的雏鸭肝脏样本中的RA，表明该检测方法可用于RA感染的临床诊断。
