Effect of foscarnet induction treatment on quantitation of human cytomegalovirus (HCMV) DNA in peripheral blood polymorphonuclear leukocytes and aqueous humor of AIDS patients with HCMV retinitis. The Italian Foscarnet Study Group

Abstract

The aim of this study was to investigate peripheral blood polymorphonuclear leukocytes and, whenever possible, aqueous humor from 65 AIDS patients with ophthalmoscopically diagnosed human cytomegalovirus (HCMV) retinitis to determine (i) whether patients consistently carry viral DNA and (ii) to what extent foscarnet induction treatment decreases viral DNA levels. HCMV DNA was quantified by PCR using densitometric analysis of hybridization products obtained from external standards and a standard curve from which the number of genome equivalents of test samples, normalized by using an internal amplification control, was interpolated. Results showed that 56 of 65 patients (86.1%) were positive for HCMV DNA prior to induction treatment. Of 41 of the 56 patients (73.2%) whose blood had become DNA negative after induction, only 5 had a high viral load (>5,000 genome equivalents per 2 x 10i polymorphonuclear leukocytes) prior to induction, whereas as many as 13 of the 15 (26.8%) patients remaining DNA positive after induction had a high viral load prior to induction. Finally, of the nine patients (13.8%) with DNA-negative blood prior to induction treatment, three were shifted to foscarnet from ganciclovir, while six were erroneously enrolled in the study. Pre-and postinduction aqueous humor samples were obtained from 12 patients; all of these were DNA positive prior to induction, whereas after induction, 4 became negative, 6 showed a marked decrease in viral DNA, and 2 had neariy stable low DNA levels. In conclusion, PCR is a valuable tool for etiologic diagnosis and monitoring of HCMV retinitis treatment in AIDS patients. HCMV DNA is consistently present in the blood and aqueous humor of all patients with HCMV retinitis. Foscarnet induction treatment is highly effective in suppressing or reducing DNA levels in both blood leukocytes and aqueous humor.Induction regimens of both foscarnet and ganciclovir have been shown to be highly effective in halting progression of human cytomegalovirus (HCMV) retinitis in patients with AIDS (2, 4, 10-15, 17, 21

Clinical Utility of Viral Load in Management of Cytomegalovirus Infection after Solid Organ Transplantation

“…To confirm the clinical suspicion of tissue-invasive CMV disease in these nonviremic situations, one may repeat the NAT at a later time point, use another NAT assay with higher sensitivity or a different genetic target, or obtain tissue for histopathologic diagnosis (54). In such cases, one should strongly consider sampling body fluid or tissue from the suspected site of infection, such as the aqueous and vitreous humor fluid for patients suspected to have CMV retinitis (139,194), BAL fluid for patients suspected to have CMV pneumonia (142,143), and CSF for patients suspected to have CMV encephalitis, meningitis, or polyradiculopathy (147,195,196).…”

Diagnosis and Management of Human Cytomegalovirus Infection in the Mother, Fetus, and Newborn Infant

“…In immunocompromised patients, HCMV DNA quantifica- In addition, it is useful for the diagnosis and local evaluation of the effect of antiviral treatment at special body sites, such as the eye and nervous system (84,210). Finally a special application concerns its use for prenatal diagnosis of HCMV infection and for quantification of viral DNA in amniotic fluid samples (see below).…”

“…To this purpose, two major approaches have been used, PCR and hybridization techniques. For PCR, two main types of competitors have been used in the quantitative-competitive PCR: homologous competitors containing small deletions or insertions with respect to the target sequence (27,76), and heterologous competitors having the target sequence for primers as the target nucleic acid but differing in the intervening sequence (84,90,95). In addition to in-house-developed methods, a commercially available method has been developed by Roche (Cobas Amplicor CMV monitor test; Roche Molecular Systems, Branchburg, N.J.) for both detection and quantification of HCMV DNA (55,128).…”

New Strategies for Prevention and Therapy of Cytomegalovirus Infection and Disease in Solid-Organ Transplant Recipients

“…While plasma PCR may be of clinical utility, further evaluation is needed before firm recommendations can be made on its applicability. PCR can also be of clinical value when analyzing urine (78,121), bronchoalveolar lavage fluid (240,448), cerebrospinal fluid (70,170,491,499), aqueous and vitreous humor samples (133,149,304), tissue biopsy samples (48,54,112,268,501), and other miscellaneous body fluids (259).…”