Xing Yuenan, Chen Ye, Feng Chengcheng, Bao Jie, Li Xiaodong, Jiang Hongbo
Aquaculture Department, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China.
J Fungi (Basel). 2023 Jul 27;9(8):791. doi: 10.3390/jof9080791.
causes a "milky disease" in Chinese mitten crab, , which inflicts significant damage on the breeding industry, but there are no effective drugs for this disease. Precise detection technologies and clarification of transmission routes are now essential to prevent its occurrence. A real-time fluorescent quantitative PCR (qPCR) detection method targeting the mitochondrial cytochrome c oxidase subunit VIA (COX6A) of was developed and its sensitivity, specificity, repeatability, and application effectiveness evaluated. There was a robust linear relationship between the qPCR threshold cycle value (Ct) and copy number of the standard with a wide dynamic range. The standard curve had a correlation coefficient (R) of 0.996, amplification efficiency of 103.092%, and a lower limit of detection sensitivity of 7.6 × 10 copies/µL. The COX6A-qPCR method exhibited high specificity for the detection of , with no cross-reactivity. The intra- and inter-group variation coefficients were <1% and 2%, respectively. The qPCR exhibited superior sensitivity compared to existing detection methods, with a positivity rate of 76.67%. The content ranged from 1.0 × 10-2.7 × 10 copies/µL. The COX6A-qPCR detection technology exhibited high sensitivity, strong specificity, and excellent repeatability, enabling the accurate quantification of .
导致中华绒螯蟹出现“乳白病”，给养殖业造成重大损失，但针对这种疾病没有有效的药物。精确的检测技术和传播途径的阐明对于预防其发生至关重要。开发了一种针对[具体病原体名称未给出]线粒体细胞色素c氧化酶亚基VIA（COX6A）的实时荧光定量PCR（qPCR）检测方法，并评估了其灵敏度、特异性、重复性和应用效果。qPCR阈值循环值（Ct）与标准品拷贝数之间存在稳健的线性关系，动态范围宽。标准曲线的相关系数（R）为0.996，扩增效率为103.092%，检测灵敏度下限为7.6×10拷贝/微升。COX6A-qPCR方法对[具体病原体名称未给出]的检测具有高特异性，无交叉反应。组内和组间变异系数分别<1%和2%。与现有检测方法相比，qPCR表现出更高的灵敏度，阳性率为76.67%。[具体病原体名称未给出]含量范围为1.0×10 - 2.7×10拷贝/微升。COX6A-qPCR检测技术具有高灵敏度、强特异性和出色的重复性，能够对[具体病原体名称未给出]进行准确定量。
