Hebei Medical University, Shijiazhuang 050031, Hebei, China; Hebei General Hospital, Shijiazhuang 050070, Hebei, China.
Hebei Medical University, Shijiazhuang 050031, Hebei, China; Hebei General Hospital, Shijiazhuang 050070, Hebei, China; NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
J Microbiol Methods. 2022 Feb;193:106404. doi: 10.1016/j.mimet.2021.106404. Epub 2022 Jan 3.
Vibrio parahaemolyticus (V. parahaemolyticus) is a widely distributed pathogen in the coastal areas, which causes food poisoning and leads to gastroenteritis and sepsis. Therefore, developing a simple, sensitive, and rapid detection method for V. parahaemolyticus is a major concern globally. This study established a sensitive and rapid technique based on recombinase aided amplification (RAA) to detect V. parahaemolyticus. The RAA reaction was carried out successfully at 39 °C within 30 min. The sensitivity of the RAA assay was 10 copies/μL using the recombinant plasmid and 10 ng/μL using the V. parahaemolyticus strain. In addition, RAA directly detected 7 × 10 CFU/mL of simulated fecal samples and 0.1 CFU/mL after enrichment for 4 h. The sensitivity and specificity of the RAA assay using fecal and fish samples were 100% similar to that of the real-time PCR. We conclude that the RAA assay is an ideal screening method for detecting V. parahaemolyticus due to its rapidity, high accuracy, and simplicity in operation.
副溶血性弧菌（Vibrio parahaemolyticus）是一种广泛分布于沿海地区的病原体，可引起食物中毒，导致肠胃炎和败血症。因此，开发一种简单、灵敏、快速的副溶血性弧菌检测方法是全球关注的焦点。本研究建立了一种基于重组酶辅助扩增（RAA）的灵敏快速检测副溶血性弧菌的技术。RAA 反应在 39°C 下 30 分钟内成功进行。使用重组质粒，RAA 检测的灵敏度为 10 拷贝/μL，使用副溶血性弧菌菌株的灵敏度为 10ng/μL。此外，RAA 可直接检测模拟粪便样本中 7×10 CFU/mL 的浓度，富集 4 小时后可检测到 0.1 CFU/mL 的浓度。RAA 检测粪便和鱼类样本的灵敏度和特异性与实时 PCR 相似，均为 100%。综上所述，由于 RAA 检测法具有快速、准确和操作简单的特点，因此是一种理想的副溶血性弧菌筛选方法。
