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不同抗原和DNA杂交方法在非洲猪瘟病毒检测中的敏感性评估。

Evaluation of sensitivity of different antigen and DNA-hybridization methods in African swine fever virus detection.

作者信息

Pastor M J, Escribano J M

机构信息

Department of Animal Health, National Institute for Agrarian Research, Madrid, Spain.

出版信息

J Virol Methods. 1990 Apr;28(1):67-77. doi: 10.1016/0166-0934(90)90088-w.

Abstract

ELISA, immunodot and DNA hybridization methods have been adapted to detect African swine fever virus (ASFV), and their sensitivities were compared using virus obtained from cell cultures. About 2.3 x 10(2) 50% hemadsorbing doses (HAD50) of virus were detected with ELISA sandwich using an anti-ASFV IgG biotinylated followed by avidin-peroxidase. The immunodot technique showed similar sensitivity, detecting about 4.6 x 10(2) HAD50 of virus. ASFV-DNA was detected using radioactive DNA probes and molecular hybridization. The maximal viral detection capacity of this technique was about 1.8 x 10(3) HAD50. The antigenic and DNA detection of ASFV during the infection of animals with virulent and attenuated viruses, was also studied. For this purpose, sera and red blood cells from several infected pigs were obtained at different days post-inoculation. The virus was detected at the third day after infection by the three methods. However, ASFV-DNA detection was more efficient than antigenic detection at nine days post-inoculation, when antigen detection failed, because immunocomplexes with circulating viruses were formed in the subacute infection.

摘要

酶联免疫吸附测定(ELISA)、免疫斑点法和DNA杂交法已被用于检测非洲猪瘟病毒(ASFV),并使用从细胞培养物中获得的病毒对它们的敏感性进行了比较。使用生物素化的抗ASFV IgG,随后是抗生物素蛋白-过氧化物酶,通过夹心ELISA检测到约2.3×10²个50%血细胞吸附剂量(HAD50)的病毒。免疫斑点技术显示出相似的敏感性,检测到约4.6×10²个HAD50的病毒。使用放射性DNA探针和分子杂交检测ASFV-DNA。该技术的最大病毒检测能力约为1.8××10³个HAD50。还研究了用强毒和弱毒病毒感染动物期间ASFV的抗原和DNA检测。为此,在接种后不同天数从几只感染猪获取血清和红细胞。感染后第三天通过这三种方法检测到病毒。然而,在接种后九天,当抗原检测失败时,ASFV-DNA检测比抗原检测更有效,因为在亚急性感染中形成了与循环病毒的免疫复合物。

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