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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™
Author(s) -
Tarragó David,
Mateos MaríaLuisa,
Avellón Ana,
PérezVázquez MaríaDolores,
Tenorio Antonio
Publication year - 2004
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.10538
Subject(s) - virology , betaherpesvirinae , polymerase chain reaction , cytomegalovirus , nested polymerase chain reaction , cerebrospinal fluid , human cytomegalovirus , herpesviridae , viral disease , real time polymerase chain reaction , biology , dna , virus , microbiology and biotechnology , pathology , medicine , biochemistry , genetics , gene
Abstract 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™ (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. J. Med. Virol. 72:249–256, 2004. © 2004 Wiley‐Liss, Inc.