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感染鸡羽毛中马立克氏病病毒抗原和DNA的检测

Detection of Marek's disease virus antigens and DNA in feathers from infected chickens.

作者信息

Davidson I, Maray T, Malkinson M, Becker Y

出版信息

J Virol Methods. 1986 Jun;13(3):231-44. doi: 10.1016/0166-0934(86)90017-0.

Abstract

Two novel tests, enzyme-linked immunosorbent assay (ELISA) and dot-blot hybridization, were developed to detect and quantify the antigens and DNA of Marek's disease virus (MDV) in feather tips from infected chickens. In both methods, buffered extracts of the feathers served as the same test material. The ELISA technique was compared to the conventional agar-gel precipitation (AGP) test, using the same convalescent serum from a MDV-infected bird. Of 86 feather samples tested, 34 were negative by both methods, while 6 out of 52 were ELISA positive but AGP negative. Viral antigen detection by the AGP and ELISA methods was compared with the detection of MDV DNA by the dot-blot DNA hybridization technique. At an ELISA reading (OD 405) of 0.3 and above, only 5 out of 48 DNA extracts failed to hybridize with the MDV-DNA probe. The use of the radioactively labelled MDV-DNA probe for hybridization with DNA extracts from feather tips of MDV-infected chickens was both sensitive and specific, and there was good correlation among the different tests.

摘要

开发了两种新型检测方法,即酶联免疫吸附测定(ELISA)和斑点杂交,用于检测和定量感染鸡羽毛尖端马立克氏病病毒(MDV)的抗原和DNA。在这两种方法中,羽毛的缓冲提取物用作相同的检测材料。使用来自MDV感染鸡的相同恢复期血清,将ELISA技术与传统的琼脂凝胶沉淀(AGP)试验进行比较。在测试的86个羽毛样本中,两种方法均为阴性的有34个,而在52个样本中,有6个ELISA阳性但AGP阴性。将AGP和ELISA方法的病毒抗原检测与斑点杂交DNA杂交技术检测MDV DNA进行了比较。在ELISA读数(OD 405)为0.3及以上时,48个DNA提取物中只有5个未能与MDV-DNA探针杂交。使用放射性标记的MDV-DNA探针与MDV感染鸡羽毛尖端的DNA提取物杂交既灵敏又特异,并且不同检测方法之间具有良好的相关性。

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