Quantitation of cytomegalovirus DNA in cerebrospinal fluid and serum specimens from AIDS patients using a novel highly sensitive nested competitive PCR and the cobas amplicor CMV monitor.
A novel nested quantitative-competitive polymerase chain reaction (nQC-PCR) assay was developed to quantify as few as ten copies per tube of human cytomegalovirus DNA with an overall dynamic range of 10-10(5) copies per tube. This nQC-PCR assay is based on co-amplification of a mimic DNA and it was evaluated with 26 cerebrospinal fluid (CSF) specimens and 44 serum specimens from 70 CMV-infected AIDS patients, 35 of them were diagnosed of CMV retinitis. An excellent correlation was found between nQC-PCR assay and the commercially available Cobas Amplicor CMV Monitor trade mark (CACM) assay (R = 0.9999; P < 0.001; n = 42). Moreover, 13 serum samples with CMV viral loads undetectable with the CACM were successfully quantified by nQC-PCR. CMV viral load was significantly higher in patients with CMV retinitis (P = 0.003). The nQC-PCR assay described below is a very sensitive test for accurate quantitative detection of CMV DNA in different clinical specimens that avoids the need for high-cost instrumentation.
Tarragó
David
D
Bacteriology Department, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain. davtarrago@isciii.es
Mateos
María-Luisa
ML
Avellón
Ana
A
Pérez-Vázquez
María-Dolores
MD
Tenorio
Antonio
A
D017088:AIDS-Related Opportunistic Infections / Q000821:virology*; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D003586:Cytomegalovirus Infections / Q000821:virology*; D017726:Cytomegalovirus Retinitis / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood* / Q000134:cerebrospinal fluid*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D019562:Viral Load
D017931:DNA Primers; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic