大黄鱼锥体虫PCR检测方法的建立与应用OA

To achieve effective prevention and control of the trypanosomosis and reduce economic losses,two pairs of specific primers were designed,and a PCR detection method based on the conserved sequence of the trypanosome 18S rRNA gene in Larimichthys crocea was established.The accuracy and sensitivity of the method were systematically evaluated through primer specificity verification,sensitivity testing,and practical sample detection.The results showed that this method produced specific amplification only for trypanosome DNA from L.crocea,with no cross-reactivity observed for non-target pathogens such as scuticociliate,Pseudo-monas putida,iridovirus,or DNA from healthy L.crocea tissues,demonstrating good specificity.The minimum detection limits of F1/R1 and F2/R2 for trypanosome were 6.52×10-39 and 6.85×10-37 ng/L,respectively,indicating high detection sensitivity.Among the 10 L.crocea samples tested,this method detected 9 positive samples,significantly outperforming the microscopy method(which detected 7 positives),further confirming its accuracy and sensitivity in practical sample detection.