An estimator of the mutant frequency in assays using transgenic animals.
The Poisson distribution is a fundamental probability model for count data, and is a natural model for the observed plaque counts in mutation assays using animals with lambda or PhiX174 transgenes. The Poisson likelihood for observed counts is a function of the mutant fraction, and it is straightforward to derive the associated maximum likelihood estimate of the mutant fraction and its variance. The estimate is easy to calculate, and if not the same, very similar to ad hoc estimates in current use. The model indicates the proper way to combine data from a number of plates, possibly prepared with different sample dilutions. The estimator of the mutant fraction is biased as a consequence of dividing by a random variable, the plaque count used to calculate the total recovered plaque-forming units. Fortunately, the bias becomes negligible as this count becomes large. On the other hand, increasing this count can increase the variance by decreasing the amount of sample assayed for mutant phages. Concurrent heed to the bias and the variance provides some guidance as to the optimum allocation of a sample into portions assayed for mutant phages and total recovered phages. The distribution of the estimate of the mutant fraction is related to the binomial distribution. This relationship implies a binomial distribution for the mutant count conditional on an overall count (either the sum of mutant and counted total plaques or the sum of counted mutant and non-mutant plaques). A special but important case occurs when each plate can be evaluated for mutant plaques and non-mutant plaques. Then, the observed proportion of mutants estimates the mutant fraction. More generally, the relationship to a binomial distribution provides a procedure for calculating a confidence interval.
Delongchamp
R R
RR
Division of Biometry and Risk Assessment, HFT-20, National Center for Toxicological Research, 3900 NCTR Rd., Jefferson, AR 72079, USA. rdelongchamp@nctr.fda.gov
Malling
H V
HV
Chen
J B
JB
Heflich
R H
RH
D000818:Animals; D001435:Bacteriophages / Q000235:genetics; D016010:Binomial Distribution; D016001:Confidence Intervals; D016013:Likelihood Functions; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D008954:Models, Biological; D009152:Mutagenicity Tests / Q000706:statistics & numerical data*; D009154:Mutation; D016012:Poisson Distribution; D011336:Probability