sybr-green-i and AIDS-Related-Opportunistic-Infections

The aim of this study is to assess a real-time PCR technique on the LightCycler 2.0 with SYBR-Green I detection as compared to another real-time PCR method based on detection with FRET (fluorescence resonance energy transfer) probes for the quantification of CMV DNA.. The two real-time PCR methods were used to test plasma samples from immunocompromised patients with clinically suspected CMV disease, patients under follow-up without symptoms, and healthy adults. A standard curve for quantitative analysis by the SYBR-Green I method was performed with 10-fold diluted solutions of DNA from the CMV Towne strain (ATCC VR-977) cultured in MRC-5 monolayer. In addition, frozen samples from patients positive for CMV pp65 antigenemia were also analyzed and results compared using the two real time PCR methods.. The real-time PCR technique using SYBR-Green I on the LightCycler 2.0 was a highly specific, fast, simple and reliable test to quantify CMV; moreover, it was cost-effective.. Quantification of CMV DNA in plasma using this sensitive, fast, low-cost method was advantageous for the diagnosis and follow up of patients with opportunistic CMV infection, which are increasingly more frequent in our daily hospital clinical practice.

Topics: Adult; AIDS-Related Opportunistic Infections; Antigens, Viral; Benzothiazoles; Bone Marrow Transplantation; Cell Line; Child; Computer Systems; Cytomegalovirus; Cytomegalovirus Infections; Diamines; DNA, Viral; Fluorescence Resonance Energy Transfer; Fluorescent Dyes; Humans; Immunocompromised Host; Infant, Newborn; Neoplasms; Neutrophils; Opportunistic Infections; Organic Chemicals; Phosphoproteins; Polymerase Chain Reaction; Postoperative Complications; Quinolines; Reagent Kits, Diagnostic; Reference Standards; Sensitivity and Specificity; Viral Matrix Proteins; Viremia