title: Detection of cytomegalovirus DNA on dried blood spots collected from infants infected with HIV: an in-house method adaptable in resource-limited settings.
abstract: In countries with limited resources, infants infected with HIV are highly exposed to CMV co-infection which probably represents a major risk factor for disease progression in this population. This study aimed to evaluate the performance of a low cost CMV DNA extraction method from DBS and the feasibility of its implementation in laboratories of 4 countries with limited resources. DNA was extracted from DBS with a cationic resin (chelex 100) and amplified with an "in house" real time CMV PCR. Dilutions of a quantified whole blood sample were spotted on paper to evaluate the 95% detection limit. A DBS quality control panel was analyzed in all laboratories. CMV PCR was compared between DBS and liquid whole blood (gold standard) in 2 populations: 418 transplanted patients and 59 infants infected with HIV (median age of 2 months). The CMV PCR 95% detection limit in DBS was 3.87 log10 copies/mL. Its positive and negative predictive values for CMV diagnosis in infants infected with HIV were 100% and 87.5% respectively. Quality control panels gave consistent qualitative results in all laboratories. This assay had high predictive values for CMV diagnosis in infants infected with HIV and its implementation in resource-limited countries with limited resources is feasible.
authors.affiliation: Virology Laboratory, Hôpital Necker Enfants malades, APHP, Université Paris Descartes, Sorbonne Paris Cité-EA 36-20, Paris, France. Electronic address: marianne.leruez@nck.aphp.fr.
authors.forename: Marianne
authors.initials: M
authors.lastname: Leruez-Ville
authors.forename: Sopeak
authors.initials: S
authors.lastname: Ngin
authors.forename: Tiffany
authors.initials: T
authors.lastname: Guilleminot
authors.forename: Anfumbom
authors.initials: A
authors.lastname: Kfutwah
authors.forename: Sandrine
authors.initials: S
authors.lastname: Moussa
authors.forename: Ton
authors.initials: T
authors.lastname: Tran
authors.forename: Eric
authors.initials: E
authors.lastname: Nerrienet
mesh_terms: D001769:Blood / Q000821:virology*; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D003890:Desiccation; D003906:Developing Countries; D015658:HIV Infections / Q000150:complications*; D006801:Humans; D007223:Infant; D016133:Polymerase Chain Reaction / Q000379:methods; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: CMV; DBS; HIV; Resource-limited countries