Cytomegalovirus DNA In Cerebrospinal Fluid Research Articles

A nested primer set targeting the cytomegalovirus glycoprotein B gene.

As the primer nomenclature may suggest to some readers, this was the fifth set we had evaluated in the search for a polymerase chain reaction (PCR) assay that could be reliably used for the laboratory diagnosis of cytomegalovirus (CMV) infections. The primer set was originally described by clinical virologists at the John Radcliffe Hospital in Oxford, UK (Read et al., 1997). They had pioneered a molecular approach to diagnosing virus infections in the central nervous system and used this primer set for CMV. Not unexpectedly their publication reported only a small number of patients (three) where the primers had detected CMV DNA in cerebrospinal fluid; two of the patients had Acquired Immunodeficiency Syndrome. However on their recommendation we undertook a wider evaluation of this primer set’s performance on clinical specimens. Following an initial 12 month period of testing specimens by both our routine assays and the nested PCR we were happy to move over entirely to the molecular method, which we continue to the present. 2. Analytical sensitivity

CMV DNA in blood and CSF of HIV infected patients

Cytomegalovirus (CMV) infection frequently affects the central nervous system in HIV infected patients. Varied clinical manifestations of CMV disease make virological detection of CMV essential for proper diagnosis and treatment. Thus, in patients in different stages of HIV-induced disease, we attempted to detect cell associated and free, non cell associated CMV DNA in cerebrospinal fluid (CSF) and in peripheral blood mononuclear cells (PBMCs). Twenty-six blood samples were collected from 22 patients included in the study. Nine of these blood samples were tested in pair with the concomitant CSF sample for the presence of CMV DNA by a commercial hybridization test. CMV serostatus and avidity of IgG antibodies were detected by a commercial ELISA test. CMV DNA was present in the cells found in CSF in all but one of the AIDS patients, independently of the presence of neurological symptoms, suggesting that it represented a marker of advanced immunodeficiency, rather than of the specific CMV-related disease. Cell-associated CMV DNA in CSF tested positive even in the samples negative for cell-free CMV DNA in the CSF, and with no detectable CMV DNA in the PBMCs of concomitant blood sample. We believe that searching for CMV DNA in different compartments of CSF merits further attention.

Detection of Herpesvirus DNA by Nested Polymerase Chain Reaction in Cerebrospinal Fluid of Human Immunodeficiency Virus-Infected Persons with Neurologic Disease: A Prospective Evaluation

A nested polymerase chain reaction-based method was used prospectively to detect herpesvirus DNA in cerebrospinal fluid (CSF) from 111 patients with AIDS, 39 of whom had a suspected diagnosis of cytomegalovirus (CMV)-associated neurologic disease (patients with encephalopathy, polyradiculopathy, or peripheral neuropathy) and 72 who had alternative diagnoses. CSF from 24 (62%) of the patients with suspected CMV-associated disease had detectable CMV DNA compared with only 8 (11%) of the patients with other diagnoses. Varicella-zoster virus DNA was detected in CSF from 3 patients (2 with myelitis and 1 with encephalitis), all of whom had recent cutaneous zoster. No CSF specimen contained detectable herpes simplex virus type 1 DNA, and none of the patients with myelitis had detectable herpes simplex virus type 2 DNA in CSF. This study demonstrates a significant association between detectable CMV DNA in CSF and suspected CMV-associated neurologic disease in patients with AIDS.

Level of Cytomegalovirus (CMV) DNA in Cerebrospinal Fluid of Subjects with AIDS and CMV Infection of the Central Nervous System

The objective of this study was to correlate cytomegalovirus (CMV) DNA levels in the cerebrospinal fluid (CSF) of subjects with AIDS with clinical and pathologic findings attributable to CMV infection of the central nervous system (CNS). CMV polymerase chain reaction (PCR) was done on serial dilutions of CSF samples from 24 AIDS patients with autopsy-proven CNS disorders. CMV DNA was detected in CSF from 12 of 13 subjects with evidence of CMV infection of the brain or spinal cord but in none of 11 subjects without autopsy evidence of CMV CNS infection. Subjects whose CSF contained > 10(3) CMV DNA molecules/8 microL of CSF had severe CMV CNS disease (e.g., ventriculoencephalitis). PCR appears to be more useful than clinical and neuroradiologic findings for documenting CMV infection of the CNS in patients with AIDS. Quantitation of CMV DNA in CSF shows promise for evaluation of the extent of involvement.

Detection of cytomegalovirus DNA in cerebrospinal fluid in immunocompetent patients as a sign of active infection.

Detection of cytomegalovirus (CMV) DNA by the polymerase chain reaction (PCR) in samples of cerebrospinal fluid (CSF) has been shown to be a sensitive method of diagnosing CMV disease in the central nervous system. Since CMV causes latent infection in white blood cells, an unanswered question is whether detection of latent CMV DNA in the cell fraction of CSF samples by PCR is possible in seropositive patients. In a prospective study, the finding of CMV DNA in CSF of CMV seropositive patients with suspected viral infection of the central nervous system (CNS) was evaluated clinically. Fractionation of 64 CSF samples from seropositive patients was carried out before analysing the samples for CMV DNA by PCR. In four of the five patients who had CMV DNA in the cell pellet and/or supernatant, the clinical data suggested CMV-associated neurological disease. The remaining 59 samples were negative in both pellet and supernatant. In addition, 11 CSF samples with high cell counts from patients with bacterial meningitis were examined for CMV DNA and found to be negative in 10 patients and positive in 1. One hundred thirty two uncentrifuged CSF samples were used as negative controls. The results of the study indicate that detection of CMV DNA in CSF samples by PCR correlated well with disease and was not due to latent CMV infection.

A prospective evaluation of clinical criteria and polymerase chain reaction assay of cerebrospinal fluid for the diagnosis of cytomegalovirus-related neurological diseases during AIDS

To study the predictive value of clinical criteria and polymerase chain reaction (PCR) assay of cerebrospinal fluid (CSF) for the diagnosis of cytomegalovirus (CMV)-related neurological disorders during AIDS. Four infectious diseases departments in two tertiary referral teaching hospitals in Paris, France. One-year prospective study involving 164 consecutive immunosuppressed HIV-seropositive patients undergoing lumbar puncture (LP). A tentative diagnostic classification, based on strict operational criteria and PCR assay of CSF, was performed at the time of LP. At the end of the study, tentative diagnoses and PCR results were blindly and independently compared with the firm diagnoses, based on central nervous system histology, clinical outcome and/or viral culture of CSF. The tentative diagnosis showed CMV-related neurological disease in 38 patients, and CMV DNA was detected in 42. Among the 88 patients for whom a firm diagnosis was possible, 26 had a diagnosis of CMV-related neurological disease. The concordance between the tentative and firm diagnoses was 61%, with a kappa index of 0.40. In contrast, the sensitivity and specificity of PCR were respectively 92 and 94%, with positive and negative predictive values of 86 and 97%. The presence of CMV DNA in CSF was associated with an increased risk of death (P < 0.0001). Unlike clinical criteria, PCR detection of viral DNA in CSF can be used reliably for antemortem diagnosis of CMV-related neurological disease, a frequent complication of AIDS in this study. This rapid method should make a major impact on the management of these patients.

A prospective evaluation of clinical criteria and polymerase chain reaction assay of cerebrospinal fluid for the diagnosis of cytomegalovirus-related neurological diseases during AIDS

Objective: To study the predictive value of clinical criteria and polymerase chain reaction (PCR) assay of cerebrospinal fluid (CSF) for the diagnosis of cytomegalovirus (CMV)-related neurological disorders during AIDS. Setting: Four infectious diseases departments in two tertiary referral teaching hospitals in Paris, France. Design and participants: One-year prospective study involving 164 consecutive immunosuppressed HIV-seropositive patients undergoing lumbar puncture (LP). Methods: A tentative diagnostic classification, based on strict operational criteria and PCR assay of CSF, was performed at the time of LP. At the end of the study, tentative diagnoses and PCR results were blindly and independently compared with the firm diagnoses, based on central nervous system histology, clinical outcome and/or viral culture of CSF. Results: The tentative diagnosis showed CMV-related neurological disease in 38 patients, and CMV DNA was detected in 42. Among the 88 patients for whom a firm diagnosis was possible, 26 had a diagnosis of CMV-related neurological disease. The concordance between the tentative and firm diagnoses was 61%, with a kappa index of 0.40. In contrast, the sensitivity and specificity of PCR were respectively 92 and 94%, with positive and negative predictive values of 86 and 97%. The presence of CMV DNA in CSF was associated with an increased risk of death (P < 0.0001). Conclusions: Unlike clinical criteria, PCR detection of viral DNA in CSF can be used reliably for antemortem diagnosis of CMV-related neurological disease, a frequent complication of AIDS in this study. This rapid method should make a major impact on the management of these patients.

Detection of Cytomegalovirus in Cerebrospinal Fluid Autopsy Specimens from AIDS Patients

Cytomegalovirus (CMV) is a common opportunistic pathogen infecting AIDS patients. Polymerase chain reaction (PCR) and antigen capture ELISA were used to detect CMV in 40 cerebrospinal fluid autopsy specimens from patients with AIDS. CMV DNA was detected by PCR in 70% of samples. Of the 21 samples from patients with systemic CMV infection, 57% had CMV encephalitis, while 81% had virus in cerebrospinal fluid detectable by PCR. Of the 24 samples from patients with no histologic evidence of CMV encephalitis, 58% had CMV DNA in cerebrospinal fluid detected by PCR. These results suggest that PCR of cerebrospinal fluid sensitively detects systemic CMV infection but is not specific for brain infection in autopsy specimens of AIDS patients.

Cytomegalovirus infection of the central nervous system in patients with AIDS: diagnosis by DNA amplification from cerebrospinal fluid.

A nested polymerase chain reaction (PCR) was evaluated for the detection of cytomegalovirus (CMV) DNA in cerebrospinal fluid (CSF). CSF and serum samples from 19 AIDS patients with intracerebral CMV infection diagnosed at autopsy were retrospectively examined. As controls, CSF and serum samples from 15 AIDS patients with only extracerebral CMV involvement at autopsy, from 10 AIDS patients without CMV infection at autopsy, and from 10 anti-human immunodeficiency virus-negative patients without ongoing CMV infection, were studied. CMV DNA was detected from patients with intracerebral CMV infection in 9 of 9, 5 of 6, and 1 of 4 CSF samples collected, respectively, 1-30, 30-90, and 90-300 days before death. Twelve of 13 sera from these patients were CMV PCR-positive. None of the control patients had CMV DNA in CSF. PCR was positive in 6 of 8 sera from AIDS patients with only extracerebral CMV infection and in serum from 1 AIDS patient without CMV involvement at autopsy. CMV PCR on CSF is highly sensitive and specific. It should be considered a rapid and reliable diagnostic method for CMV infection of the central nervous system.

Rapid detection of cytomegalovirus DNA in cerebrospinal fluid of AIDS patients with neurologic disorders.

A polymerase chain reaction (PCR)-based method was used to detect cytomegalovirus (CMV) DNA in 82 cerebrospinal fluid (CSF) samples from 67 patients infected by human immunodeficiency virus (HIV). The test was positive for 14 patients, 8 of whom had CMV-related neurologic disease proven by viral culture of CSF or histologic examination. Encephalitis was the most frequent manifestation in patients with positive PCR results, but CMV DNA was also present in some patients with peripheral neuropathy or polyradiculomyelitis. All patients with proven CMV neurologic disease were positive by PCR. In contrast, viral culture was negative for 4 of the 8 patients and pathologic studies were available only for 5. The specificity of the PCR-based assay could not be assessed precisely because of the lack of a reference standard, but the results correlated well with clinical course and results of the other methods. These findings suggest that the PCR-based method may be a useful noninvasive tool for the rapid diagnosis of CMV-related neurologic disease.