Detection of Cytomegalovirus DNA in Peripheral Blood of Patients Infected with Human Immunodeficiency Virus

Abstract

Detection of cytomegalovirus (CMV) DNA can be facilitated by the polymerase chain reaction (PCR), an in vitro gene amplification technique. Twenty-eight CMV tissue culture isolates were examined by amplification of two separate CMV genes. All were found to contain CMV, although two of the isolates were positive for only one of the two genes. No detectable amplification occurred with human genomic or other viral DNA controls. The amplification products from as few as one CMV plaque-forming unit could be detected after the PCR. CMV DNA was detected in the blood of 14 of 27 patients with AIDS and one of six patients who were infected with human immunodeficiency virus but who did not have AIDS. Normal CMV-seropositive or -seronegative individuals did not have CMV DNA detected in their blood. The CMV PCR was more sensitive than the standard culture assay, can be completed in one to two days, uses only 20 microL of blood, and may be useful for rapidly detecting CMV in clinical specimens.

Prevalence of sexually transmissible pathogens in semen from asymptomatic male infertility patients with and without leukocytospermia

“…Oligonucleotide sequences for CMV, HHV-6, HSV-1/2 (together) and EBV were obtained from the literature (39)(40)(41)(42)(43), the sequences for HBV were obtained from Boehringer Mannheim (Penzberg, Germany). The oligonucleotides were synthesized by Genosys (The Woodlands, TX, USA).…”

Quantitative determination of human cytomegalovirus target sequences in peripheral blood leukocytes by nested polymerase chain reaction and temperature gradient gel electrophoresis

“…Thus, during acute infection with human cytomegalovirus (HCMV), viral DNA is readily detected by PCR in peripheral blood mononuclear cells (PBMCs), biopsies, serum, urine and various other specimens (Brytting et al, 1992;Demmler et al, 1988;Jiwa et al, 1989a, b;Rogers et al, 1990;Shibata et al, 1988), but positive results may be obtained during asymptomatic reactivations and in latently infected healthy individuals as well (Bevan et at., 1991). Therefore, without further laboratory and clinical data, positive PCR results are difficult to interpret.…”

Comparison of serology, antigenemia assay and the polymerase chain reaction for monitoring active cytomegalovirus infections in hematopoietic stem cell transplantation patients

“…Since CMV infections are either asymptomatic or associated with nonspecific clinical manifestations, laboratory techniques are essential for diagnosing the viral infection. PCR and AGM are considered the most effective markers for diagnosing active CMV infection in transplant recipients 1,2,5,[9][10][11][12]14,16 . In BMT recipients, the ability of PCR to detect active CMV infection may be better than that of AGM 1,2 .…”

“…The slides were air-dried and fixed with formaldehyde, prior to immunostaining with monoclonal antibodies C10 and C11 (Clonab CMV; Biotest, Dreieich, Germany), and then reacted with peroxidase-labeled antimouse conjugate (HRP, Biotest, Dreieich, Germany). CMV-DNA in blood specimens was detected by nested-PCR using primers described by DEMMLER et al 3 and SHIBATA et al 14 . Briefly, leukocytes remaining from the CMV-AGM assay were lysed and the DNA was precipitated.…”

“…In recent years, there have been important advances in antiviral prophylactic strategies and in the development of sensitive diagnostic techniques for monitoring recipients of bone marrow transplantation (BMT) 2 . Newer methods, such as early CMV pp65 antigenemia assay and CMV-DNA amplification, can diagnose CMV infection in its very early period 3,14 . Serological tests for detecting specific IgM antibodies and/or a significant increase in specific IgG concentrations are valuable diagnostic tools in BMT recipients with primary or recurrent CMV infection 4,6 .…”