Cunningham D A, Pattison J R, Craig R K
Department of Biochemistry, University College, London, U.K.
J Virol Methods. 1988 Mar-Apr;19(3-4):279-88. doi: 10.1016/0166-0934(88)90022-5.
Methods were established for the detection of parvovirus DNA in human serum using single-stranded RNA probes. The sensitivity of detection of virus using 32P-radiolabelled RNA versus non-radiolabelled biotinylated RNA probes using a streptavidin-polyalkaline phosphatase detection system was compared. Virus was detected using 32P-labelled and biotinylated RNA probes at serum dilutions of 10(-3), equivalent to approx. 3 pg of viral DNA. Using biotinylated RNA probes and a dot-blot system, diagnosis of numerous serum samples could be performed within 8 h of receipt of samples, using an RNA probe which was synthesised and stored at -20 degrees C for up to 12 months without loss of sensitivity. Our work demonstrates the potential of biotinylated RNA probes in the routine analysis of viral sequences in serum.
建立了使用单链RNA探针检测人血清中细小病毒DNA的方法。比较了使用32P放射性标记的RNA与使用链霉亲和素-多碱性磷酸酶检测系统的非放射性生物素化RNA探针检测病毒的灵敏度。在相当于约3 pg病毒DNA的10(-3)血清稀释度下，使用32P标记和生物素化的RNA探针检测病毒。使用生物素化的RNA探针和斑点印迹系统，在收到样品后的8小时内可以对大量血清样品进行诊断，使用的RNA探针在-20℃合成并储存长达12个月，灵敏度无损失。我们的工作证明了生物素化RNA探针在血清病毒序列常规分析中的潜力。
