PMID: 9171217Jun 10, 1997Paper

Quantitation of target molecules from polymerase chain reaction-based limiting dilution assays

AIDS Research and Human Retroviruses
Allen G RodrigoJ I Mullins

Abstract

Polymerase chain reaction-based limiting dilution assays (PLDAs), commonly called end-point dilutions, are frequently used to quantify the copy numbers of human immunodeficiency virus (HIV) and other viruses in biological samples; however, the way in which these assays are done, and the mathematical method used to estimate copy numbers, vary from laboratory to laboratory. Here, we describe a statistical method for estimating the number of copies and the associated standard error of the estimate, using a PLDA. The copy number is estimated by the value that maximizes the goodness of fit between the observed numbers of negative reactions and the expected numbers of negative reactions (the latter estimated using a Poisson probability distribution) as measured by the chi2 statistic. The method described here also takes into account user-specified probabilities of obtaining a false-positive or a false-negative PCR result, a feature that is not generally available with other limiting dilution estimation procedures. QUALITY, a computer program that implements the estimation strategy, is also described. Simulations illustrate the efficiency of estimation with different numbers of PCR amplifications conducted at each dilution, and differ...Continue Reading

References

Dec 14, 1989·The New England Journal of Medicine·D D HoM Alam
Jul 26, 1996·Science·S L LiuJ I Mullins

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Citations

Jul 22, 1998·Journal of Immunological Methods·K B Walker
Jan 19, 2000·Immunology Today·I DozmorovI Lefkovits
Apr 21, 2004·The Journal of Molecular Diagnostics : JMD·Roberto Flores-MunguiaAnna R Giuliano
Apr 7, 2005·European Journal of Human Genetics : EJHG·Huanliang LiuTuofu Zhu
Mar 21, 2002·Scandinavian Journal of Immunology·A S GoldmanF C Schmalstieg
Apr 14, 2010·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Mark A MicekLisa M Frenkel
Oct 27, 2007·AIDS Research and Human Retroviruses·Gonzalo BelloCecilio López-Galíndez
Apr 20, 2004·AIDS·Patricia N Fultz
Sep 29, 2006·Journal of Clinical Microbiology·Greg BrennanSue VandeWoude
Sep 28, 2012·Journal of Virology·Denis R ChoperaUNKNOWN CAPRISA 002 Study Team
Aug 28, 2004·Journal of Virology·Alexandra CochranePeter Simmonds
Dec 28, 2005·Journal of Virology·Rosario SabariegosMiguel Angel Martínez
Jun 19, 2008·Virology Journal·Jessica J Hayward, Allen G Rodrigo
Mar 15, 2011·PloS One·Mary S CampbellUNKNOWN Partners in Prevention HSV/HIV Transmission Study Team
Oct 8, 2013·PloS One·Brendan B LarsenJames I Mullins

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