Heldt Caryn L, Hernandez Raquel, Mudiganti Usharani, Gurgel Patrick V, Brown Dennis T, Carbonell Ruben G
Department of Chemical and Biomolecular Engineering, North Carolina State University, 911 Partners Way, Raleigh NC 27695-7905, USA.
J Virol Methods. 2006 Jul;135(1):56-65. doi: 10.1016/j.jviromet.2006.01.022. Epub 2006 Mar 6.
Many animal viruses produce cytopathic effects in their host cells during a productive infection. While some virus infections can be assayed by the production of plaques, many viruses, while producing cytotoxicity, do not easily form plaques, or do not form plaques at all. Additionally, viruses within families such as the parvoviruses may have different preferred forms of titration making comparative virology difficult even among related groups. Porcine parvovirus (PPV), canine parvovirus (CPV), and minute virus of mice (MVM) are usually titrated using different infectivity assays. A direct comparison of infectious virus titer between these parvoviruses was sought, and a tetrazolium salt assay, MTT has been applied to measure cytopathic effect produced by viral infection for different members of the parvovirus family. Infectious PPV measured using the MTT and the TCID50 assays exhibited excellent correlation and titers for CPV and MVM were consistently duplicated using the MTT assay. The MTT assay was also applied to an unrelated virus, Sindbis, which is routinely titrated by plaque assay. MTT titration of Sindbis virus mutants was found to be valuable for preliminary screening. This assay can be adapted, by correlation to an accepted titration method, to any viral system which produces measurable cytopathic effect.
许多动物病毒在增殖性感染期间会在其宿主细胞中产生细胞病变效应。虽然一些病毒感染可以通过噬斑的产生来检测,但许多病毒虽然产生细胞毒性,却不容易形成噬斑,或者根本不形成噬斑。此外,细小病毒科等家族中的病毒可能有不同的首选滴定形式,这使得即使在相关病毒组之间进行比较病毒学研究也很困难。猪细小病毒(PPV)、犬细小病毒(CPV)和小鼠微小病毒(MVM)通常使用不同的感染性检测方法进行滴定。本研究旨在直接比较这些细小病毒之间的感染性病毒滴度,并应用一种四唑盐检测法(MTT)来测量细小病毒家族不同成员病毒感染产生的细胞病变效应。使用MTT和TCID50检测法测得的感染性PPV显示出极好的相关性,并且使用MTT检测法能一致地重复测得CPV和MVM的滴度。MTT检测法还应用于一种不相关的病毒——辛德毕斯病毒,该病毒通常通过噬斑检测法进行滴定。发现辛德毕斯病毒突变体的MTT滴定法对于初步筛选很有价值。通过与一种公认的滴定方法建立相关性,该检测法可适用于任何产生可测量细胞病变效应的病毒系统。