鸽圆环病毒可视化LAMP检测方法的建立与应用OA北大核心

Primers were designed targeting the highly conserved region of pigeon circovirus(PiCV)Rep gene,cresol red was used as an indicator,and a visual loop-mediated isothermal amplification(LAMP)method was established for PiCV detection after condition optimization and specificity and sensitivity evaluation.Then thirty-five clinical samples of pigeons were detected using the established method and the conventional PCR method.It showed that the detection result could be determined by a visual color change of the reaction liquid after reacting at 64 ℃ for 60 min and then at 85 ℃ for 10 min.The reaction liquid of the positive reaction tube changed from purple-red to orange,and the color of the negative reaction tube remained unchanged.No cross-reactivity was found in the detection of pigeon adenovirus,pi-geon herpesvirus,pigeonpox virus,and pigeon paramyxovirus Ⅰ by the established method.The lowest de-tection limit was 17.4 copies/μL recombinant plasmid,indicating its sensitivity was 100 times that of the conventional PCR method.Sixteen positive samples were detected by the visual LAMP method,while 15 were detected by the conventional PCR method.It indicates that the visual LAMP method established in this study can be applied to clinical sample detection,which provides technical support for convenient and rapid diagnosis of pigeon circovirus infection.