Comparison of HCMV loads using in-house quantitative CMV PCR, Artus CMV TM PCR and COBAS Amplicor CMV monitor test.

Abstract

A real-time PCR assay based on highly conserved cytomegalovirus glycoprotein B (CMV-gB) gene was developed to quantify the CMV genome loads in human plasma obtained from immunocompromised patients. The sequence comparison from several strains resulted in the best possible set of primers and a probe that directed against the most highly conserved region. The detection limit of In-house TaqMan polymerase chain reaction assay for CMV-DNA was 2.3 log10 copies/ml and the linear measure interval was 2.3 to 8.3 log10 copies/ml (R > 0.99). The In-house TaqMan assay was compared to commercial quantitative PCR test, COBAS Amplicor CMV monitor test and Artus CMV TM PCR test, in the determination of CMV DNA load in plasma samples. The correlation between In-house TaqMan and COBAS assays was statistically significant, R = 0.915 (P < 0.05; n = 27). Also correlation between In-house TaqMan and Artus CMV assays was statistically significant and higher (R = 0.998; P < 0.05; n = 8). When the In-house TaqMan system was evaluated with the Nucleospin ® RNA virus kit and QIAamp blood minikit, no significant difference was found at the 98.2% confidence interval. Preliminary results indicate that the In-house TaqMan real-time PCR with the designed primer and probe set can be an alternative tool for quantification of CMV loads in clinical routine laboratories.