The method described in this chapter uses limiting dilution analysis in conjunction with RT-PCR to determine quantitatively what percentage of EBV-infected cells within a given population are expressing the viral genes EBNA-1 Q-K, EBNA-2, LMP-1, LMP-2, BZLF-1, and the EBERs. Because this technique involves limiting dilution analysis, it is possible to define which viral transcription programs a...

Limiting dilution assays are commonly used to measure the extent of infection, and in the context of HIV they represent an essential tool for studying latency and potential curative strategies. To assist investigators using dilution assays, we illustrate the major statistical method for estimating the frequency of infected cells (or other infectious units) from assay results, and we offer an on...

AIMS To obtain floating spheres from the adherent gastric cancer cell line SGC-7901 and to analyze the properties of the spheres. METHODS Serum-free medium was applied to cultured SGC-7901 cells. Limiting dilution assay, tumor formation assay, microarray analysis, and real-time fluorescent quantitative PCR were used to test the stem cell properties of the spheres. RESULTS A subpopulation of...

Digital polymerase chain reaction (PCR) is an emerging absolute quantification method based on the limiting dilution principle and end-point PCR. This methodology provides high flexibility in assay design without influencing quantitative accuracy. This article describes an assay to quantify HIV DNA that targets a highly conserved region of the HIV-1 genome that hampers optimal probe design. To ...

Preliminary studies have suggested that depletion of T lymphocytes from donor marrow might be an effective method for preventing acute graft-v-host disease (GVHD) after allogeneic bone marrow transplantation in humans (Lancet 1:369, 472, 1984). However, the minimum degree of T cell depletion required to assure the prevention of GVHD in a population of marrow graft recipients has not been define...