title: Quantitative HIV-1 proviral DNA detection: a multicentre analysis.
abstract: Despite the widespread use of molecular biology techniques, standardized methods for the measurement of HIV-1 proviral DNA are currently lacking and several discordant results are still present in different studies. To assess the clinical meaning of the proviral DNA load, a study group comprising seven different laboratories was set up to standardize a HIV-1 proviral DNA quantification method able to assess the DNA proviral load of the most relevant circulating HIV-1 subtypes. Reference samples (24 cellular samples infected with HIV-1 clade B, and 40 samples of peripheral blood mononuclear cells containing different concentrations of plasmids expressing different HIV-1 clades) were distributed and tested blindly. All laboratories employed hTERT gene as housekeeping gene and primers within the gag gene to quantify different HIV-1 clades. Inter-laboratory results did not differ statistically but showed only minor variations concerning HIV-1 DNA amounts and different HIV clades, with a good agreement among the laboratories participating in the study. Since test standardization represents a key step for future application in clinical practice, further studies of the patients' samples are in progress to establish the real meaning and utility of the proviral DNA load for clinical management of HIV-1 infected patients.
authors.affiliation: Department of Oncology and Surgical Sciences, Oncology Section, Unit of Viral Oncology, AIDS Reference Centre, University of Padova, IOV-IRCCS, Padova, Italy.
authors.forename: Anita
authors.initials: A
authors.lastname: De Rossi
authors.forename: Marisa
authors.initials: M
authors.lastname: Zanchetta
authors.forename: Francesca
authors.initials: F
authors.lastname: Vitone
authors.forename: Guido
authors.initials: G
authors.lastname: Antonelli
authors.forename: Patrizia
authors.initials: P
authors.lastname: Bagnarelli
authors.forename: Luigi
authors.initials: L
authors.lastname: Buonaguro
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: Capobianchi
authors.forename: Massimo
authors.initials: M
authors.lastname: Clementi
authors.forename: Isabella
authors.initials: I
authors.lastname: Abbate
authors.forename: Filippo
authors.initials: F
authors.lastname: Canducci
authors.forename: Alessia
authors.initials: A
authors.lastname: Monachetti
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Riva
authors.forename: Gabriella
authors.initials: G
authors.lastname: Rozera
authors.forename: Carolina
authors.initials: C
authors.lastname: Scagnolari
authors.forename: Maria
authors.initials: M
authors.lastname: Tagliamonte
authors.forename: Maria Carla
authors.initials: MC
authors.lastname: Re
mesh_terms: D002460:Cell Line; D004279:DNA, Viral / Q000032:analysis*; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007558:Italy; D016133:Polymerase Chain Reaction / Q000592:standards*; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000235:genetics; D019562:Viral Load / Q000592:standards*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000235:genetics
chemical_list: D004279:DNA, Viral; D054301:gag Gene Products, Human Immunodeficiency Virus; C509186:TERT protein, human; D019098:Telomerase