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Fungal infections of the central nervous system (CNS) are rare but they pose a significant challenge. Their prevalence spans a wide array of hosts including immunosuppressed and immunocompetent individuals, patients undergoing neurosurgical procedures and those carrying implantable CNS devices. Cryptococcus neoformans and Aspergillus spp. remain the most common pathogens. Magnetic resonance imaging can help localize the lesions, but diagnosis is challenging since invasive procedures may be needed for the retrieval of tissue, especially in cases of fungal abscesses. Antigen and antibody tests are available and approved for use in the cerebrospinal fluid (CSF). PCR-based techniques are promising but they are not validated for use in the CSF. This review provides an overview on the differential diagnosis of the fungal CNS disease based on the host and the clinical syndrome and suggests the optimal use of diagnostic techniques. It also summarizes the emergence of Cryptococcus gatti and an unanticipated outbreak caused by Exserohilum rostratum.

The involvement of the central nervous system in dengue infections has been reported in countries where the disease in endemic. The purpose of this study was to determine whether an enzyme-linked immunosorbent assay kit designed to detect the dengue NS1 antigen in serum was able to detect this antigen in cerebral spinal fluid (CSF) samples from patients with fatal outcomes. To evaluate the sensitivity of the kit, 26 dengue-positive CSF samples were used. The Pan-E Dengue Early kit was able to detect the NS1 antigen in 13 of 26 dengue-positive CSF samples, resulting in a sensitivity of 50% (95% confidence interval, 29.9-70.1%) and specificity of 100% (95% confidence interval, 75.3-100%). The kit was able to detect the NS1 antigen in CSF of individuals who had died of dengue. When used in combination with IgM, the detection rate rose to 92.3%. This study reports a method for rapidly detecting the dengue virus in CSF, thereby increasing the diagnosis of dengue fever cases with unusual neurological manifestations., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Herpes simplex encephalitis (HSE) represents one of the most severe infectious diseases of the central nervous system. As effective antiviral drugs are available, early rapid and reliable diagnosis has become important. The objective of the present study was to develop a sensitive enzyme-linked immunosorbent assay (ELISA) protocol for herpes simplex virus (HSV) antigen detection by assessing the usefulness of hyperimmune sera isolated from HSV-seropositive patients. A total of 52 cerebrospinal fluid (CSF) and 62 serum samples of HSE patients and non-HSE persons were analyzed. An in-house ELISA protocol utilizing hyperimmune sera was developed for HSV antigen detection. To improve the specificity of the method, protein A was incorporated into the protocol for ELISA. The sensitivity (70% and 90%) of antigen detection was high in CSF and serum samples, respectively, of confirmed HSE patients. However, lower specificity (52.3% and 42.3%), respectively, was obtained, which was improved by using protein A in the ELISA protocol. The modification in the method yielded good sensitivity (80% and 70%) and specificity (85.7% and 88.4%) of HSV antigen detection in the CSF and sera, respectively, of the HSE and non-HSE groups. The ELISA method utilizing hyperimmune sera along with protein A for HSV antigen detection yielded good sensitivity and specificity in both CSF and sera, and hence can be useful for the diagnosis of HSE.

The resurgence of chikungunya (CHIK) in the form of unprecedented explosive epidemic after a gap of 3 decades in India and Indian Ocean islands is a point of major public health concern. The laboratory diagnosis is essentially based on virus isolation, IgM ELISA, and reverse transcriptase polymerase chain reaction (RT-PCR). Although PCR-based methods are used for early and accurate diagnosis, the high cost of the assay and requirement of thermal cycler limit its application only to referral laboratories. The antibody-based IgM ELISA is found to be cost-effective, but it takes 5 to 6 days for the patient to develop antibody and, thus, has less implication for early clinical diagnosis and patient management. Therefore, a simple rapid, sensitive, and specific antigen detection system is reported for early and reliable clinical diagnosis as well as effective surveillance of CHIK. A double antibody sandwich system was designed for antigen capture ELISA, employing rabbit and mouse anti-CHIK IgG antibodies as capture and detector antibodies, respectively. An optimal assay condition with 0 background was established having no reactivity with healthy human serum and Cerebro spinal fluid (CSF) samples. The comparative evaluation with SYBR Green I-based real-time RT-PCR revealed an accordance of 96% with a sensitivity and specificity of 95% and 97%, respectively. The specificity of this assay was confirmed through cross-reactivity studies with confirmed dengue and Japanese encephalitis (JE) patient serum and CSF samples. The antigen capture ELISA reported in this study was able to detect the presence of viral antigen as early as the second day of fever and, thus, can be very useful for early clinical diagnosis of CHIK with acute phase patient serum and CSF samples. This can also be used for rapid screening of large numbers of clinical samples in endemic areas during epidemics.

Group A rotavirus (GARV) genes (the VP7 and NSP3 genes) in acute-phase cerebrospinal fluid (CSF), sera and stool samples from 6 children with convulsions accompanied by GARV gastroenteritis were investigated by reverse transcription-polymerase chain reaction (RT-PCR). When the VP7 gene was amplified from the samples, the G genotype (G type) of GARV was determined by RT-PCR. GARV genes were detected in the CSF samples of all 6 children, in 2 of the 3 blood samples, and in all of 4 stool samples. The G typing of GARV from 12 of a total of 13 samples indicated that G3 was the predominant G type in all samples. GARV antigens were detected by enzyme-linked immunosorbent assay in all of the 3 tested sera samples, while no GARV antigens were detected in any of the 5 tested CSF samples. We confirmed the presence of GARV genomes in the CSF samples from all of the children with rotavirus-associated seizures, including encephalopathy. However, the relationship between convulsions and the existence of GARV RNA in CSF remains unclear and further study is required.

Infection with human T-cell lymphotropic virus type I (HTLV-I) have been associated with the development of the HTLV-I-associated myelopathy/Tropical Spastic Paraparesis (HAM/TSP). The disease affects the pyramidal tract at the distal segments of spinal cord, generating a spastic paraparesis. We studied the presence of Tax protein in cerebrospinal fluid cells and spinal fluid (CSF) of 35 Chilean patients: 22 HAM/TSP patients (15 HTLV-I-seropositives, and 7 seronegatives), and 13 controls (9 PSP and 4 CJD non-infected patients). Tax antigens were evaluated with monoclonal antibodies reacting with Tax by immunofluorescence and ELISA assays in cerebrospinal fluid cells and CSF, respectively. Proviral was evaluated by PCR of tax gene in cerebrospinal fluid cells. Tax antigen was detected in CSF and lymphocytes of CSF from 4 and 12 HAM/TSP patients, respectively. Lymphocytes of CSF of 8 HAM/TSP (6 seropositives and 2 seronegatives) showed the presence of tax gene. These results show that cells of CSF from HAM/TSP patients are able to express and export Tax protein towards the CSF. This is the first report of the presence of Tax protein in cerebrospinal fluid cells and CSF from HAM/TSP HTLV-I seronegative patients.

We report a case of transverse myelitis as a complication of acute cytomegalovirus (CMV) infection in immunocompetent patients; and review the literature on the entity. Primary CMV infection was documented by CMV antigenemia and high serum titers of CMV IgM and IgG antibodies. Cerebrospinal fluid (CSF) pleocytosis indicated central nervous system inflammation; CSF polymerase chain reaction (PCR) for CMV, however, was negative. The results of magnetic resonance imaging of the myelon were normal. Although CMV-associated transverse myelitis has been well described in HIV-positive individuals, but is very rare in immunocompetent individuals. It remains unclear whether the neuronal damage is immune mediated or due to a cytotoxic effect of viral infection. The outcome is mainly favorable.

Objectives: A retrospective study on cerebrospinal fluid (CSF) samples was made possible by access to a large CSF bank, which has been supplemented by the well characterised prior collection of CSF from patients with Herpes simplex encephalitis (including serial samples from some patients and kindly donated by Professor Maurice Longson, Manchester). These samples are of particular interest because they were collected prior to the routine administration of acyclovir., Methods: Although an earlier study had shown that there was indeed a correlation between higher titres of antibody and a better outcome, the data did not emerge with statistical significance. The current study was based upon an improved method, which demonstrates the antigen-specific clonality of the immune response., Results: The primitive polyclonal anamnestic response was contrasted with the strong antigen-specific response, which was manifested by a monoclonal pattern in some patients. A clear distinction emerged between two sub-groups of patients on the basis of these findings, which showed a statistically significant (P<0.03) correlation with outcome., Conclusions: This has allowed us to further support the hypothesis that a strong immunological response has positive prognostic value during the course of the disease.

Multiple sclerosis (MS) is thought by many investigators to have an infectious component, and several microorganisms have been associated with the disease during the last three decades. Recent studies have implicated both human herpesvirus 6 (HHV-6) and the obligate intracellular bacterium Chlamydia pneumoniae in the etiology of MS. As with earlier studies of other potential agents, however, evidence linking either of these organisms to the disease is equivocal. In this article, we review data for and against involvement of HHV-6 and C. pneumoniae in MS, as well as evidence concerning auxiliary factors, such as possession of the APOE epsilon4 allele, which may influence the role of these organisms in pathogenesis. Further, we suggest several lines of investigation that should clarify whether either or both pathogens are associated meaningfully with this disease.

Eight newborn calves showing ataxia were necropsied and examined histologically. Six of seven cerebrospinal fluid samples collected from these animals had neutralizing antibody for Akabane virus (AKV). All examined calves had nonsuppurative encephalomyelitis, localized mainly in the midbrain and spinal cord. Corresponding to the encephalitic lesion, AKV antigen was demonstrated in neuroglial cells in the brain stem and neuronal cells in the ventral horn of the spinal cord. This is the first study to demonstrate AKV antigen by immunohistochemistry in naturally infected newborn calves.

The role of the viral burden in the brain for the pathogenesis of human immunodeficiency virus-associated neurological disorders is still unclear. To address this issue, we have quantified the viral load in plasma, cerebrospinal fluid (CSF) and brain tissue of macaques infected with simian immunodeficiency virus (SIV). We discovered that the viral strain used for infection determines the replicative capacity in microglial cells as well as the extent of neuropathological lesions and the occurrence of neurological symptoms. Moreover, the viral load in the brain parenchyma correlated with the development of overt neurological disease whereas the one in plasma did not. By comparing the viral load in three different compartments, we demonstrated that the viral burden in the CSF is influenced both by the viral replication in the periphery as well as in the brain parenchyma. According to these results, it is not the absolute amount of viral load in the CSF but rather the viral antigen contributed by the viral production within the brain which correlates with the development of neurological disease. In longitudinal studies, we observed that this autochthonous virus production, as evidenced by a ratio of the viral load in CSF to the one in plasma, takes place for a prolonged period of time before overt neurological signs are manifested. This finding suggests that this ratio could be used as a prognostic marker for immunodeficiency virus-induced neurological disease.

Two features of the biology of JC virus make it a particularly suitable candidate for an agent in MS-like disease: its neurotropic capability targeting glial cells as evidenced in progressive multifocal leukoencephalopathy lesions, and its capacity for latency and persistence as illustrated by its behaviour in the kidney. JC virus is chronically or intermittently excreted in the urine by some 40% of the population. The existence of JC virus in multiple coding-region genotypes provides a unique approach to the study of JC virus-induced neurological disease. We have previously shown that a genotype originating in Asia but also present in Europe and the US, called Type 2B, is more frequently found in PML brain than expected based on its prevalence in urine samples from a control population. In contrast, we find that the excretion of JCV in MS patients is similar in both genotype and frequency to that of control individuals, and appears to be regulated by factors unrelated to those that control CNS disease activity.

Objective: To investigate the relation between biochemical alterations and disease severity in HIV-cognitive motor complex (HIV-CMC)., Background: HIV-CMC encompasses both the milder form (HIV-minor cognitive motor disorder [HIV-MCMD]) and the more severe form (HIV-dementia). There is no validated marker to monitor disease severity noninvasively., Methods: A total of 54 patients with HIV-CMC (20 with HIV-MCMD, 34 with HIV-dementia) and 29 seronegative healthy volunteers were evaluated for cerebral metabolite abnormalities using proton (1H) MRS in the frontal cortex, frontal white matter, and basal ganglia., Results: The three subject groups showed different concentrations of myoinositol (MI; p = 0.0005) and choline-containing compounds (CHO; p = 0.004) in the frontal white matter. HIV-dementia patients had metabolite changes in all three brain regions whereas HIV-MCMD patients had abnormalities in the frontal white matter only. HIV-CMC patients had elevated MI (p < 0.0001) and CHO (p = 0.004) levels with increasing AIDS dementia complex stage, and N-acetyl compounds (NA) were decreased only in moderate to severe stages of dementia. Furthermore, CD4 count and CSF viral load, but not plasma viral load, showed significant effects on cerebral metabolite concentrations, which in turn showed significant effects on the HIV-dementia scale., Conclusions: In early stages of HIV-CMC, frontal white matter showed evidence of glial proliferation (with elevated MI and CHO levels) and cell membrane injury (with increased CHO levels), but no significant neuronal injury (with normal NA concentrations). HIV-MCMD and HIV-dementia patients have different neurochemical abnormalities. Because these biochemical alterations are related to clinical disease severity, they may be useful surrogate markers for noninvasive quantitative assessment of brain injury in patients with HIV-CMC.

Cerebrospinal fluid (CSF) obtained from 20 clinically healthy dogs and from 15 dogs affected by neurological disorders were examined for total and differential cell counts, immunocytochemistry for canine distemper virus antigen, total protein concentration and electrophoretic separation, and glucose and enzyme determination. Dogs affected by canine distemper showed an increase in macrophages, presence of specific inclusion bodies, and an increase in total protein concentration and gamma-globulin fraction. In bacterial meningoencephalomyelitis pleocytosis, increase of total proteins with special regard to high molecular weight proteins, elevated levels of lactate dehydrogenase (LDH), alterations of LDH isoenzyme profile, and decrease in glucose levels were detected; inflammatory disorders were more often characterized by an increase in LDH level, while in non-inflammatory disorders (hydrocephalus and spinal cord neoplasia) no variation in LDH levels was detected. Analysis of CSF in dogs appears relatively easy to perform and may help in establishing the condition of the blood-brain barrier as well as in the diagnosis of neurological disorders.

Branched chain DNA assay (bDNA), cytomegalovirus (CMV) antigen assay, and cerebrospinal fluid (CSF) viral culture were studied for their utility in the diagnosis of CMV polyradiculopathy and for documenting in vivo antiviral effects. CMV was demonstrated in 15 of 16 patients by bDNA assay, 15 of 16 by CMV antigen assay, and 11 of 15 by CSF culture. When clinical criteria and results of the other two assays were used as reference standards, the sensitivity of bDNA was 94% and 100% and the specificity 95.2% and 100%; the CMV antigen assay sensitivity was 94% and 100% and specificity was 85.7% and 100%. Nine (90%) of 10 patients with polyradiculopathy and follow-up CSF culture showed a drop in CMV DNA after treatment; however, only 2 (20%) improved clinically. These results suggest that bDNA and antigen assays may be useful methods for the diagnosis of CMV polyradiculopathy, but treatment failures may not be due to inadequate antiviral activity.

Encephalomyelitis is a sequela to ovine lentivirus (OvLV) and human immunodeficiency virus infections. Examination of autopsy tissue from 38 naturally infected asymptomatic sheep showed that 7 (18%) had subclinical neurological lesions characterized by perivascular and periventricular infiltrates of lymphocytes and histiocytes in the leptomeninges, cerebral white matter, choroid plexus, and/or cervical spinal cord. Intralesional histiocytes were shown to contain lentiviral capsid proteins or RNA. Infectious virus (2/7), viral proteins (4/7), and antiviral antibody (5/7) were only detected in cerebrospinal fluid (CSF) from animals with central nervous system (CNS) lesions associated with OvLV infection, suggesting that such virologic markers in CSF, when used concurrently, are predictive of pathologic changes specific to the CNS.

We report five children who had recurrent central nervous system signs after conventional acyclovir therapy for herpes simplex encephalitis. Secondary exacerbation was characterized clinically by severe ballismic movement disorder in all five children, associated with fever, impairment of consciousness, and seizures. Biologic analysis in all children and magnetic resonance imaging and neuropathology studies of the brain in three cases were compatible with inflammatory reaction. In contrast, all viral cultures remained negative, herpes simplex virus antigen in one child and DNA tested by polymerase chain reaction in four children were undetectable in the first samples of cerebrospinal fluid during the relapse, suggesting a postinfectious, immune-mediated mechanism of relapse in these patients.

Painful peripheral neuropathy (PPN) in HIV-infected patients has been increasingly associated with cytomegalovirus (CMV) infection at other sites. In the last few years, the detection of CMV lower matrix phosphoprotein (pp65) antigen in leukocytes has become a major tool in the diagnosis of CMV systemic infection in immunocompromised patients. In this study, CMV antigen detection was assessed in 13 HIV-infected patients with PPN and, as controls, in 82 HIV seropositive patients without any evidence of peripheral nerve syndromes (10 with CMV retinitis and 72 without CMV end-organ disease). CMV antigenemia was found in 10 (76.9%) patients with PPN, in 5 (6.9%) without CMV disease, and in all 10 patients (100%) with CMV retinitis. Of the 10 PPN patients with CMV antigenemia, only 3 presented with CMV retinitis, while the remaining 7 had no clinical evidence of overt CMV infection at other sites. CMV pp65-positive cells were also found in three of the four cerebrospinal fluid (CSF) samples collected from PPN patients. Ganciclovir was effective in improving neurological symptoms in two of the four treated patients. The findings suggest that active CMV infection may be associated with PPN in HIV infection even in the absence of CMV disease at other sites. The detection of CMV-matrix pp65 antigen in the blood and CSF leukocytes could represent a simple and rapid tool of selecting PPN patients for antiviral therapy.

A technique for detecting herpes simplex virus (HSV) antigen in CSF by measuring the antigen-antibody complex-activated complement-stimulated chemiluminescence in normal granulocytes was applied to 17 CSF samples from 10 patients with herpes simplex virus encephalitis (HSVE), and 26 samples from 22 patients with a non-HSVE brain illness. All CSF samples taken from the 5th to the 38th day (including seven from the 5th to the 10th day) of HSVE were positive, whereas those for all non-HSVE samples were negative. This assay required only five hours to yield a result.

Subacute sclerosing panencephalitis (SSPE) is related to "defective" measles virus or vaccination, though an association with parainfluenza viruses has been reported. SSPE is characterized by a slow, erratic course and elevated cerebrospinal fluid measles titers. An immunocompetent, vaccinated infant, with onset of symptoms in parainfluenza virus season and a catastrophic course is described. Cerebrospinal fluid titers were negative, but postmortem brain had typical SSPE lesions. Patient brain-derived RNA, subjected to reverse transcription followed by polymerase chain reaction yielded polymerase chain reaction products with measles virus but not parainfluenza virus genes. The sequenced fragment revealed multiple mutations, typical for SSPE. SSPE can thus present in infants, with short latency and no cerebrospinal fluid antibodies. Viral genomic analysis may be diagnostic, permitting early therapy.

The polymerase chain reaction (PCR) was used to detect DNA of herpes simplex virus (HSV) in 38 samples of cerebrospinal fluid (CSF) obtained from 22 patients (7 children and 15 adults) 1-20 days after the onset of encephalitis. The results were best with amplification on the pellet of the CSF-extracted DNA and with analysis of the amplified products by dot-blotting (sensitivity, 100%). A highly significant difference was evident in the chi 2 test when PCR was compared with specific antigen detection or antibody evaluation (n = 19; chi 2 = 7; sensitivity = 100% vs. 63%) or with interferon alpha determination (n = 20; chi 2 = 11; sensitivity = 95% vs. 42%). PCR was positive as early as 1 day after onset of disease and was often the first test to become positive. The detection of HSV DNA by PCR is the most specific, rapid, and sensitive tool for early diagnosis and therapeutic management of acute HSV encephalitis.

This paper reports the direct evidence for the presence of Japanese encephalitis virus (JEV)-specific immune complexes in the CSF of 31/185 confirmed patients for the first time. A monoclonal antibody-based capture ELISA was used for the detection of immune complexes. Amongst the 31 cases positive for immune complexes, 14 were positive for JEV IgM antibodies and/or neutralizing antibodies in the CSF, 6 were positive for viral antigen in the CSF and 11 for both antibody and antigen. Correlation of findings to final outcome revealed that the presence of immune complexes in CSF was significantly associated with death (p = 0.01).

Continuous intravenous administration of zidovudine (AZT) has been reported to improve cognitive function in HIV-infected pediatric patients (Pizzo et al., 1988). The effects of long-term zidovudine treatment in the perinatally infected pediatric population, including antiviral efficacy and effects on cognitive and motor function has not been systematically examined. These questions were addressed in rhesus macaque infants infected at birth with SIVSMM/B670, a primate model for infantile HIV infection and disease (Eiden et al., 1993a). Continuous or intermittent administration of AZT during the first 6 months following infection resulted in about a doubling of lifespan, a delay in the occurrence of motor impairment, and lower virus burden and quinolinic acid levels in cerebrospinal fluid (CSF) following administration of the antiviral drug.

Immunological and virological evidence for persistence of Japanese encephalitis virus (JEV) in the human nervous system is described in 16/323 (5%) laboratory-confirmed cases of Japanese encephalitis. In 9/16 patients, JEV specific IgM antibodies were detected in the CSF even at 50-180 days after the onset of symptoms. Similarly, in 7/16 patients, apart from IgM antibodies, viral antigen was also present in the CSF beyond the third week of illness and in one patient it could be detected even at 117 days. Infectious virus could be isolated from the CSF beyond the third week of illness in 3/16 patients. In one patient, JEV was isolated from the CSF on three consecutive occasions at 90, 110, and 117 days after onset of clinical symptoms. These findings suggest that JEV persists in the nervous system of a small proportion of patients.

Objectives: Infections caused by the cytomegalovirus (CMV) are frequent complications in patients with the acquired immunodeficiency syndrome. Diagnosis of CMV infections of the nervous system is still difficult., Methods: We evaluated immunocytochemical staining for a CMV early antigen in cells prepared from the cerebrospinal fluid of patients with clinical evidence for neural CMV infections and from controls., Results: CMV positive cells were found in four samples of patients with clinical evidence of CMV infection. Three of these patients were suffering from myeloradicalitis, one had a severe brain stem encephalitis. No CMV positive were found in controls., Conclusions: Immunocytochemical staining of CSF cells may be helpful in the diagnosis of CMV infection of the central nervous system and of the nerve roots.

In this study we tested the hypothesis that IgG paraproteins in neurological diseases might be regarded as an anti-idiotypic response raised by a nervous system antigen associated with certain neurotropic agents. After initial ELISA screen, positive samples from 34 neurological patients who had paraproteins in their CSF and serum on routine IEF investigation, were tested by immunoblot technique for the presence of specific monoclonal immunoglobulin G (IgG) against eight different neurotropic antigens. Only one patient had specific monoclonal IgG against herpes simplex virus. The results of this study did not confirm our hypothesis. Further study of IgM/IgA paraproteins is indicated.

Routine brain biopsy is often recommended in all patients suspected of having herpes simplex encephalitis (HSE). This approach is based upon the lack of ability to make the diagnosis of HSE on clinical grounds, the need to exclude other diseases for which there is specific therapy, and to stop empiric therapies. Acyclovir is a relatively safe, effective antiviral agent with little toxicity as currently used to treat HSE. Thus this is not a problem. Careful review of alternate diagnoses established at brain biopsy reveals few for which biopsy is the only and best way of making the diagnosis. Thus empiric therapy with acyclovir should be considered in some children in whom all appropriate diagnostic techniques fail to establish specifically treatable diseases. Early detection of HSV antigen in CSF may provide a non-invasive means of early diagnosis in the future.

Herpes simplex encephalitis (HSE) carries a high mortality rate. Therefore, an early diagnosis and institution of acyclovir are essential. We report a case of biopsy-proven HSE with 2 negative cerebrospinal fluid (CSF) analyses and 2 normal CT scans. However, MRI together with EEG were abnormal early in the disease stressing their significant role in any suspected case of HSE. Although brain biopsy remains controversial, CSF herpes simplex antigen detection offers hope in providing an early or retrospective diagnosis while specific antiviral therapy with acyclovir is initiated. Overdependency on routine CSF analysis or head CT scan can result in unnecessary delays in diagnosis and treatment.

Reference intervals are reported for feline CSF biochemical and serologic variables, IgG concentration, and electrophoretic fractionation, derived from 58 clinically normal adult cats that did not have histologic lesions of the CNS. There was no apparent effect of age on any variable. The CSF total protein concentration was significantly (P = 0.012) greater in males than in females, but all other variables were unaffected by gender. The only variable that had a statistically significant correlation with its corresponding blood concentration was IgG. Blood contamination of the CSF affected the following CSF variables: total protein concentration, activities of lactate dehydrogenase and creatine kinase, IgG ratio, and gamma-globulin percentage. The reference intervals proposed for feline CSF were derived from 33 cats with CSF RBC count less than 31 cells/microliters. Reference limits for CSF with 31 to 1,700 RBC/microliters also are reported.

Six patients with herpes simplex encephalitis were investigated. Antibody activities against different viral antigens were determined in serum and cerebrospinal fluid with an enzyme-linked immunosorbent assay after adjusting both fluids to identical immunoglobulin G concentrations. In serum, the strength of the antiviral reaction remained low and no qualitative changes became detectable. In cerebrospinal fluid, at first locally synthesized antibodies were directed against the same antigens as in serum. Ten days later, the intrathecal reaction increased with additional antibodies against at least two antigens. Once established, this expanded heterogeneity remained stable during the course of the disease.

A patient with atypical manifestations of Epstein-Barr Virus (EBV) encephalomyelitis is presented. The patient had unusual spinal fluid immunoglobulin abnormalities, the syndrome of inappropriate anti-diuretic hormone secretion, autonomic dysfunction and spinal arachnoiditis. The cisternal CSF, with a very high IgG level (34.3 mg/dl; normal less than 6.1) but a normal albumin content, displayed evidence of massive intrathecal immunoglobulin production. This was further confirmed by the presence of oligoclonal bands. These clinical manifestations have not previously been reported in encephalomyelitis associated with EBV infection.

An indirect immunofluorescent technique for rapid diagnosis of patients of Japanese encephalitis (JE) was developed by demonstrating the viral antigen in CSF cells. The CSF samples of 31 consecutive patients admitted with acute viral encephalitis during 1987 were studied. In 15 patients the diagnosis of JE virus infection was made on the same day. Classical techniques for virus detection and antibody serology were also conducted in these patients. The indirect immunofluorescence for JE virus antigen detection in CSF cells proved to be a rapid and simple procedure for making a quick diagnosis of infection.

Herpes-zoster associated encephalitis (HZAE) is an uncommon complication of herpes zoster. Over 8 years, we evaluated 12 patients with this clinical diagnosis. The majority of our patients were elderly, immunosuppressed, and found to have disseminated skin lesions prior to the onset of CNS symptoms. All patients had abnormal EEGs, and CSF pleocytosis was found in most. In the seven patients who were tested, specific antibody to the varicella-zoster membrane antigen (FAMA) was detected in spinal fluid during the course of the illness. Although three patients died during the period of active infection, the virus could not be definitively implicated as the cause of death. These HZAE patients could not be distinguished from our other herpes zoster patients on the basis of age, initially involved dermatome, or mortality rate. However, among herpes zoster patients who survived, duration of hospitalization was significantly longer in those with a diagnosis of HZAE. All surviving HZAE patients had a slow but eventual return to their prior cognitive status.

We investigated several methods for the rapid diagnosis of herpes simplex virus induced encephalitis in a rabbit model. The corneas of twenty-two rabbits were infected with herpes simplex virus type 1 (HSV-1) and diagnosis of Herpes encephalitis was made by virus isolation, immunofluorescent and peroxidase staining of brain biopsies, demonstration of anti-HSV IgM in cerebrospinal fluid (CSF) and by an indirect enzyme-linked immunosorbent assay (ELISA), designed for detection of viral antigens. With the last method we were able to demonstrate viral antigens in cerebrospinal fluid six days post infection, before clinical signs of encephalitis appeared. In three rabbits this was before anti-HSV IgM appeared in the CSF. Virus was isolated from brain samples of 67 per cent of the animals which died from Herpes encephalitis. Nine rabbits received cortisone before infection, resulting in markedly lower antibody titers and a higher lethality, 77 per cent, as compared to 46 per cent in nontreated rabbits. For rapid diagnosis of Herpes encephalitis in rabbits, demonstration of herpes simplex virus antigens in CSF by means of an indirect ELISA is superior to the other methods investigated.

Thirty-two cerebrospinal fluid (CSF) samples from eighteen patients with confirmed herpes simplex encephalitis (HSE) were assayed by an indirect enzyme-linked immunosorbent assay (ELISA) for the presence of viral antigens. The results are expressed as an antigen ratio distinguishing between herpes simplex virus (HSV) antigens containing samples and negative samples. Judged by this criterion a positive result was obtained in 33% of the patients. Overall, 25% of the CSF samples from HSE patients were positive. In one out of 33 control patients with other neurological disorders a positive antigen ratio was found. Two or more CSF samples were available from eleven patients. In six of these, the second or later samples showed a decreased antigen ratio when compared to the first CSF sample. An increase of the anti-HSV antibody titer was seen in the CSF of five of these six patients. Five out of six patients with a decreasing antigen ratio had an unfavorable outcome of their encephalitis, while a favorable outcome was seen in four of the five patients with an increasing or steady antigen ratio. A decrease of the antigen ratio in the course of HSE can be explained by the presence of immune complexes in CSF and may indicate a poor prognosis.

The diagnosis on the exclusion of infectious diseases of the central nervous system, especially of bacterial infections still is one of the most important issues in clinical microbiology. In bacterial meningitis, where lethal courses as well as severe sequelae are still frequent, there should be a rapid diagnosis not only with microscopy but also with Limulus test and antigen detection tests because a specific therapy should be initiated as soon as possible. But also viral infections caused by varicella or herpes virus are increasingly susceptible to chemotherapy. The indication for the examination of cerebrospinal fluid, the minimal volume for exhaustive laboratory tests and the possibilities of a stepwise diagnostic procedure are given with reference data from literature for the various techniques. In our experience the synopsis of laboratory results and clinical symptoms yielded in 75% of all cases the exclusion of an infectious etiology of the disease. In 17% a bacterial meningitis or the infection of a hydrocephalus shunt could be diagnosed. Viral infections could be proven in 4% either by antibody or by antigen detection. Only in 1% of all patients the clinical symptoms and the laboratory parameters remained unclear.

Immune complexes from the CSF of 12 individuals (6 with exacerbations of MS and 6 with other neurologic or psychiatric disease) were isolated and characterized. Three MS patients had complexed herpes simplex type I viral antigen and antibody; three patients had complexed myelin basic protein. Two MS patients and one with hypoxic encephalopathy had complexed antibody directed against brain glycolipids. CSF complexes were distinct from serum complexes, suggesting intrathecal origin. Reactivation of latent brain virus may play a role in exacerbations of MS.

Cerebrospinal fluid (CSF) specimens were obtained from patients with presumed herpes simplex encephalitis who underwent brain biopsy for diagnostic confirmation. Coded CSF specimens were fixed on nitrocellulose filter paper and probed with a pool of monoclonal antibodies directed against four herpes simplex virus glycoproteins (gB, gC, gD, and gE). Herpes simplex virus antigen was detected in 35 of 40 specimens obtained from 26 biopsy-positive patients. In contrast, only three of 25 specimens from 17 biopsy-negative patients gave positive results by this assay. An additional 30 CSF specimens from patients with proven bacterial and fungal infections were all negative by this assay. For all specimens tested, the sensitivity and specificity of the assay were both 88%. However, when results were evaluated by patient, the sensitivity was 92% (24 of 26) with a specificity of 82% (14 of 17). Among specimens collected one week or later after disease onset, the sensitivity was 100%, with a specificity of 93%.

An aetiological agent for multiple sclerosis has not yet been identified. The paramyxovirus SV5 (Simian virus 5) has been suggested as an important cause in some cases. Using antigen immunoblotting with SV5 virus, we confirmed the binding to SV5 of immunoglobulin G from the cerebrospinal fluid of 58% of multiple sclerosis patients, but also found binding in 70% of patients with a variety of neurological disorders where oligoclonal banding was present. This suggests that the antibodies present are not specific for multiple sclerosis and that the SV5 virus is thus unlikely to be of aetiological significance.

This article discusses general principles of laboratory diagnosis, detection of microbial antigens, specific chemical reactants associated with inflammation of the CNS, and diagnostic tests in development for the laboratory diagnosis of infectious meningitis.

Among the confirmed cases of viral meningitis and encephalitis, the most commonly diagnosed agents are enteroviruses, arboviruses, and herpes simplex virus. Definitive diagnosis of a viral infection often affects patient management, length of hospitalization, and antibiotic use. Laboratory diagnostic methods include microscopic examination of clinical specimens, virus culture, serologic studies, and immunologic detection of virus or viral antigens. Microscopic examination of cerebrospinal fluid is nonproductive, and, except for enteroviruses, culture almost always gives negative results. Viral culture of the cerebrospinal fluid, throat, and feces is the diagnostic method of choice for enteroviruses. Definitive diagnosis of herpes simplex virus encephalitis must be based on virus isolation from brain biopsy material. Arbovirus infection is diagnosed serologically. Although none of the newer rapid immunologic techniques is commercially available, some do hold great promise. These include measurement of virus-specific immunoglobulin M (IgM) and detection of viral antigens by enzyme or radioimmunoassay. A sensitive and specific procedure for early detection of herpes simplex virus antigens in cerebrospinal fluid would be of great benefit.

Mice injected intracerebrally with a sublethal dose of vaccinia virus develop severe meningitis. The number of inflammatory cells in cerebrospinal fluid (CSF) increases approximately threefold each day for 3 to 7 days after intracerebral challenge, subsequent to which samples can no longer be obtained because the cisterna magna is obliterated owing to brain swelling. Examination of this inflammatory exudate during the later stages of this pathological process shows evidence of both cytotoxic T lymphocyte (CTL) function and the presence of Lyt1+ and Lyt2+ cells on days 6 and 7. In addition, potent non-T-cell (Lyt2-) cytotoxic activity is found in CSF taken from younger (12 weeks) mice as early as after day 3 and is still present on day 6. The level of non-T-cell-mediated cytotoxicity in CSF on day 5 or 6 (but not day 3) is considerably decreased in animals that were also given a large dose of virus intravenously to maximize T-cell stimulation in lymphoid tissue, and the extent of CTL activity is concomitantly increased. The diminution of non-specific cytotoxic function does not seem to reflect simple dilution in the presence of excess virus-immune T cells.