CALIENDO A, SCHUURMAN R, YEN-LIEBERMAN B, SPECTOR SA, ANDERSEN J, CRUMPACKER C, LURAIN N, ERICE A; Interscience Conference on Antimicrobial Agents and Chemotherapy.
Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 1999 Sep 26-29; 39: 438 (abstract no. 1951).
Emory Univ. Hosp., Emory, GA
BACKGROUND: The risk of developing CMV disease in AIDS patients is directly related to the presence and quantity of CMV DNA in plasma. A variety of PCR assays are available for detection and quantitation of CMV DNA.METHODS: We analyzed the performance characteristics (lower limit of detection, inter- and intra-assay variability, linear range, upper limit of quantification) of commercial qualitative (Roche-Quant) and quantitative (Roche-Quant) PCR assays for CMV DNA. The correspondence between the Roche-Quant assay and a non-commercial quantitative PCR assay (UCSD) was also analyzed. A quantitation panel (range, 0 to 1 x 10[6 ]CMV DNA copies/mL) was prepared by diluting cell-free EM-counted AD169 in human plasma.RESULTS: Three series (1 unblinded; 2 blinded) of the quantitation panel were tested in 4 laboratories: 3 laboratories used Roche-Qual and Roche-Quant assays, and one laboratory used the UCSD assay. Replicates containing 0 CMV DNA copies/mL tested negative in all assays. The Roche-Qual assay consistently detected CMV DNA in dilutions containing 2,400 copies/mL (unblinded series) or 12,000 copies/mL (blinded series). The Roche-Quant assay consistently detected CMV DNA in dilutions with 2,400 copies/mL (unblinded and blinded series). Twelve (67%) and 3 (17%) of the replicates with dilutions containing 31 and 63 DNA copies/mL of the viral stock, respectively, had <400 CMV DNA copies/ mL in the Roche-Quant assay; CMV DNA was quantifiable in 8 (53%) of these replicates. The linear range (log scale) of the Roche-Quant assay was from 400 to 10 x 10[4] copies/mL, and that of the UCSD assay was from 780 to > 13 x 10[4] copies/mL. The intra- and inter-laboratory variability of the quantitative PCR assays was greater at the lower concentrations of the viral stock.CONCLUSIONS: The Roche-Quant PCR assay is more sensitive than the Roche-Qual PCR assay for detection of CMV DNA. This finding has practical importance for studies of the natural history of CMV as well as to identify individuals at risk for developing CMV disease in whom prophylactic intervention may be warranted.
Publication Types:
Keywords:
- Biological Assay
- Cytomegalovirus
- Cytomegalovirus Infections
- DNA
- DNA Primers
- Humans
- Polymerase Chain Reaction
- Reproducibility of Results
- Research
- analysis
- transplantation
Other ID:
UI: 102245586
From Meeting Abstracts