title: The increased sensitivity of human cytomegalovirus (HCMV) PCR quantitation in whole blood affects reproductive rate (Ro) measurement.
abstract: In order to determine the effect of the increase in sensitivity of HCMV detection in whole blood compared to plasma on reproductive rate (Ro) measurement, an optimized human cytomegalovirus (HCMV) quantitative PCR assay was developed. The results presented in this study are summarized by the following three methodological improvements: (i) at values below the limit of quantitation (LOQ) of 60copies/ml, determination of HCMV load was more sensitive with whole blood than plasma, (ii) for the determination of viral load, whole blood was more sensitive than plasma below 1000copies/ml but little difference was observed above 1000copies/ml and (iii) the measurement of "Reproductive Rate" can be affected by imprecise measurement of HCMV viral load in either plasma or whole blood compartments depending on whether samples were taken from patients on antiviral treatment or from patients where HCMV load was rising. Taken together this study provides methodological improvements suggesting that below HCMV viral load levels of 1000copies/ml (1640IU/ml) both plasma and whole blood should be tested.
authors.affiliation: Department of Pathology, Austin Hospital, Studley Road, Heidelberg 3084, Australia; School of Applied Sciences, Building 223, Level 1, Bundoora Campus RMIT University, PO Box 71, Bundoora 3083, Australia. Electronic address: volker.gurtler@gmail.com.
authors.forename: Volker
authors.initials: V
authors.lastname: Gurtler
authors.affiliation: Department of Pathology, Austin Hospital, Studley Road, Heidelberg 3084, Australia.
authors.forename: Barrie C
authors.initials: BC
authors.lastname: Mayall
authors.affiliation: Department of Pathology, Austin Hospital, Studley Road, Heidelberg 3084, Australia.
authors.forename: Jenny
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Pathology, Austin Hospital, Studley Road, Heidelberg 3084, Australia.
authors.forename: Shahbano
authors.initials: S
authors.lastname: Ghaly-Derias
mesh_terms: D050936:Basic Reproduction Number; D001769:Blood / Q000821:virology*; D003587:Cytomegalovirus / Q000254:growth & development* / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000453:epidemiology / Q000821:virology*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*
keywords: HCMV; Human cytomegalovirus; IU; LOD; LOQ; Plasma; Quantitative real-time PCR; R(o); SNP; WB; Whole blood; human cytomegalovirus; international units; limit of quantitation; lower limit of detection; reproductive rate; single nucleotide polymorphism; whole blood