Yang Limin, Li Jing, Bi Yuhai, Xu Lei, Liu Wenjun
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Virus Genes. 2012 Dec;45(3):585-9. doi: 10.1007/s11262-012-0798-6. Epub 2012 Aug 7.
We developed and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting Duck hepatitis A virus type 1 (DHAV-1). The amplification could be finished in 1 h under isothermal conditions at 63 °C by employing a set of four primers targeting the 2C gene of DHAV-1. The RT-LAMP assay showed higher sensitivity than the RT-PCR with a detection limit of 0.1 ELD(50) 0.1 ml(-1) of DHAV-1. The RT-LAMP assay was highly specific; no cross-reactivity was observed from the samples of other related viruses, bacteria, allantoic fluid of normal chicken embryos, or the livers of uninfected ducks. Thirty clinical samples were subjected to detection by RT-LAMP, RT-PCR, and virus isolation, which obtained completely consistent, positive results. As a simple, rapid, and accurate detection method, this RT-LAMP assay has important potential applications in the clinical diagnosis of DHAV-1.
我们开发并评估了一种用于检测1型鸭甲型肝炎病毒（DHAV-1）的逆转录环介导等温扩增（RT-LAMP）检测方法。通过使用一组针对DHAV-1 2C基因的四条引物，在63℃等温条件下1小时内即可完成扩增。RT-LAMP检测方法显示出比RT-PCR更高的灵敏度，检测限为0.1 ELD(50)/0.1 ml DHAV-1。RT-LAMP检测方法具有高度特异性；未观察到来自其他相关病毒、细菌、正常鸡胚尿囊液或未感染鸭肝脏样本的交叉反应。对30份临床样本进行了RT-LAMP、RT-PCR检测和病毒分离，结果完全一致且均为阳性。作为一种简单、快速且准确的检测方法，这种RT-LAMP检测方法在DHAV-1的临床诊断中具有重要的潜在应用价值。
