Liu Mei-Ling, Xia Yong, Wu Xing-Zhong, Huang Jian-Qiong, Guo Xu-Guang
Department of Clinical Laboratory Medicine, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150, China.
Department of Clinical Laboratory Medicine, Guangdong Center for Skin Disease & STI Control Guangzhou, Guangzhou, 510091, China.
AMB Express. 2017 Dec;7(1):48. doi: 10.1186/s13568-017-0349-6. Epub 2017 Feb 23.
Gonorrhea is a sexually transmitted disease caused by the bacterium Neisseria gonorrhoeae. Rapid detection is crucial for effective prevention and treatment. This study developed and tested a low-cost effective method for detecting N. gonorrhoeae, especially in developing countries.
DNA from a N. gonorrhoeae standard strain, as well as from 26 genital secretion samples of gonorrhea patients, were isolated and used for loop-mediated isothermal amplification (LAMP) assay, which was conducted using either an automatic real-time PCR analyzer or a water bath. The amplified porA pseudogene sequence was compared with the NCBI database and the LAMP results were compared with that of the traditional culture method for its sensitivity and specificity.
LAMP was able to detect Neisseria DNA at a concentration as low as 1 pg/µL (1 × 10 CFU/mL cells). The LAMP assay results obtained using an automatic real-time PCR analyzer was similar to that of the water bath. Relative to traditional culture, the sensitivity and specificity of the LAMP assay were 94.7 and 85.7%, respectively.
LAMP was sensitive and reliable for detecting the porA gene of N. gonorrhoeae. It could be used as a rapid, low cost, and effective method for detecting N. gonorrhoeae.
淋病是由淋病奈瑟菌引起的一种性传播疾病。快速检测对于有效预防和治疗至关重要。本研究开发并测试了一种低成本的有效方法来检测淋病奈瑟菌，尤其是在发展中国家。
从淋病奈瑟菌标准菌株以及26份淋病患者的生殖器分泌物样本中分离DNA，并用于环介导等温扩增（LAMP）检测，该检测使用自动实时PCR分析仪或水浴进行。将扩增的porA假基因序列与NCBI数据库进行比较，并将LAMP结果与传统培养方法的结果进行敏感性和特异性比较。
LAMP能够检测低至1 pg/µL（1×10 CFU/mL细胞）浓度的奈瑟菌DNA。使用自动实时PCR分析仪获得的LAMP检测结果与水浴结果相似。相对于传统培养，LAMP检测的敏感性和特异性分别为94.7%和85.7%。
LAMP检测淋病奈瑟菌的porA基因灵敏且可靠。它可作为一种快速、低成本且有效的淋病奈瑟菌检测方法。
