Detection of cytomegalovirus in plasma of patient’s HIV by duplex real time PCR
Keywords:
Cytomegalovirus, Polymerase Chain Reaction, Plasma, UL122, UL83Abstract
Background: Cytomegalovirus (CMV) is able to cause serious and even deadly diseases in immunocompromised patients. It is important to have a sensitive, specific and molecular viral tests for its detection, using as targets, key genes for viral replication. The following genes have been used in the molecular detection of CMV: UL122 (replication) and UL83 (most abundant protein of the tegument).
Objective: Detect and quantify CMV, by real-time duplex PCR, from a minimum amount of plasma.
Material and methods: The UL122 and UL83 genes were amplified with different fluorophores, by real-time duplex PCR. To quantify CMV, curves were generated, starting with DNA-CMV (1.0-0.0000001 ng).
Results: The dynamic range of “master” duplex straight had a pendent (m) −3.0, the amplification efficiency was 115.44% plasmas from patients with HIV viral load ≥ 100,000 copies/mL, 11.36% were true positive for CMV and 88.64% had no amplifications or they were outside of the linear range of molecular detection.
Conclusions: This test identified two important CMV genes (UL122 and UL83) in a single reaction (FAM:VIC), viral detection was confirmed from a minimum amount of plasma. This mean a smaller amount of biological sample required and would add a tool to the clinical area, as well as a lower consumption of reagents and materials.
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