In recent years, a new type of Spiroplasma has been found that can cause “tremor disease” of the Chinese mitten crab Eriocheir sinensis. The outbreak of epidemic tremor disease has caused a serious setback in the Chinese mitten crab farming industry, with an incidence rate of more than 30% and mortality rates of 80–100%. Therefore, finding a sensitive method to detect tremor disease in E. sinensis has become a current research focus. In this research, a loop-mediated isothermal amplification detection method coupled with hydroxynaphthol blue dye (LAMP-HNB) was developed and used to rapidly detect Spiroplasma eriocheiris. First, we designed and synthesized specific outer primers, inner primers and loop primers based on the 16S ribosomal RNA gene of S. eriocheiris. Second, the LAMP-HNB detection method for S. eriocheiris was successfully established by screening the primers, adjusting the temperature and time of the reaction, and optimizing the concentrations of Mg²⁺ and dNTPs. In the specific tests, only samples infected with S. eriocheiris showed positive results, and other infections caused by bacteria and parasites tested negative, proving that the test has high specificity. Moreover, the detection limit was 2.5 × 10^–6 ng/µL, indicating high sensitivity. This method for detecting S. eriocheiris provides rapid visual output based on LAMP-HNB detection and is a simple, fast, sensitive, and inexpensive method that can be applied to a wide range of field investigations.

近年来，发现一种新型螺原体，可引起中华绒螯蟹“震颤病” 。流行性震颤病的爆发，使我国大闸蟹养殖业严重受挫，发病率达30%以上，死亡率达80%～100%。因此，寻找一种灵敏的方法来检测中华石蚕震颤病已成为当前的研究热点。本研究开发了一种结合羟基萘酚蓝染料（LAMP-HNB）的环介导等温扩增检测方法，用于快速检测马氏螺原体。首先，我们基于鲫鱼16S核糖体RNA基因设计并合成了特异性的外引物、内引物和环引物。其次，通过引物筛选、反应温度和时间的调整以及Mg~ ^{(2+)和dNTPs浓度的优化，成功建立了}绒毛链球菌的LAMP-HNB检测方法。在具体检测中，只有绒毛链球菌感染的样本呈阳性，其他由细菌和寄生虫引起的感染检测结果为阴性，证明该检测具有较高的特异性。此外，检测限为2.5 × 10 ^–6  ng/μL，灵敏度较高。这种检测S. eriocheiris的方法提供基于 LAMP-HNB 检测的快速视觉输出，是一种简单、快速、灵敏且廉价的方法，可应用于广泛的现场调查。