Author:
Spira T J,Bozeman L H,Holman R C,Warfield D T,Phillips S K,Feorino P M
Abstract
A micromethod for assaying the reverse transcriptase enzyme of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus in cocultures of clinical specimens for viral isolation was developed and compared with the macromethod in use. Ultracentrifuged, pelleted, and solubilized viral culture supernatants were transferred into either tubes (macromethod) or microtiter plates (micromethod) and incubated with tritiated enzyme substrate. Trichloroacetic acid-precipitated DNA was collected on individual filter papers with a Millipore filtration manifold (macromethod) or on filter sheets using a semiautomated cell harvester (micromethod). Filters were then placed in scintillation fluid and counted on a beta scintillation counter. Results of the micromethod significantly correlated to those of the macromethod, with a linear relationship between the two. The cutoffs for positivity based on the mean + 2 standard deviations for a set of known negative specimens (n = 19) was 4,973 cpm for the micromethod compared with 5,336 for the macromethod. The intrarun and interrun variations were comparable for both methods. There was a 67% increase in the maximal daily number of specimens which could be run (100 versus 60) as well as a reduction in reagent use. In summary, the micromethod utilizing a semiautomated cell harvester is comparable to the existing macromethod in accuracy and is an improvement due to savings in time and reagents.
Publisher
American Society for Microbiology
Reference5 articles.
1. Primer requirement and template specificity of the DNA polymerase of RNA tumor viruses;Baltimore D.;Proc. Natl. Acad. Sci. USA,1971
2. Distinguishing reverse transcriptase of an RNA tumor virus from other known DNA polymerases;Goodman N. C.;Proc. Natl. Acad. Sci. USA,1971
3. Neter J. W. Wasserman and M. H. Kutner. 1985. Applied linear statistical models. Richard D. Irwin Homewood Ill.
4. Viral and cellular DNA polymerase: comparison of activities and synthetic and natural RNA templates;Robert M. S.;Science,1972
5. Snedecor G. W. and W. G. Cochran. 1980. Statistical methods 7th ed. Iowa State University Press Ames.
Cited by
79 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献