Differential diagnosis of eosinophilic meningitis

Meningitis - infection o f the meninges - can be attributed to a variety o f agents, including bacteria, protozoa and some helminths. In helminth infections, but rarely in protozoal infections, eosinophilia is a common sign. Thus eosinophilic meningitis (EOM) is typically associated with certain helminth infections in which nervous system involvement occurs. Among the most important of these are ongiostrongyliosis, gnathostomiasis, porogonimiasis and cysticercosis (see Fig. 1). Here, Nibha Jaroonvesoma discusses the key distinctions between such infections.     

The rapid differential diagnosis of bacterial and viral meningitis by using the lysozyme test]

The authors have modified the technique of the lysozyme test by adding polimixin M sulfate into the gel bacterial medium. Rapid diagnosis with the use of this test is based on different time of the appearance of the lysis areas: in bacterial meningitides the CSF lysozyme activity is detectable within 15-120 min, whereas in viral meningitides it manifests 40-50 min later or does not manifest at all. The results were found to depend on the time of the CSF collection: the earlier the CSF samples were obtained, the higher was the share of positive results.     

Coagglutination test and its application to the rapid diagnosis of meningococcal meningitis

Modified technique of slide coagglutination test for detecting meningococcal group-specific antigens in the spinal fluid of patients with meningococcal meningitis has been developed. Precipitating meningococcal sera, groups A, C, X, Y, Z, were conjugated with formalin-treated staphylococcal cells, strain Cowan-I. To prevent nonspecific reactions, 5-minute boiling of the spinal fluid specimens is suggested. 111 specimens of spinal fluid were taken from 75 patients at different periods of the disease. All patients were administered antibiotics, and therefore the etiology of the disease was bacteriologically confirmed only in 31% of patients. Coagglutination test was positive in 56.7% of patients, the frequency of positive results reaching 71% during the first 4 days of the disease. The specimens of spinal fluid taken from the control group of patients yielded not more than 2% of the positive results. Coagglutination test is recommended as a rapid test for diagnosing meningococcal meningitis.     

Molecular diagnosis of eosinophilic meningitis due to Angiostrongylus cantonensis (Nematoda: Metastrongyloidea) by polymerase chain reaction-DNA sequencing of cerebrospinal fluids of patients

Cerebrospinal fluid (CSF) samples from clinically diagnosed patients with detectable Angiostrongylus canto-nensis-specific antibodies (n = 10), patients with clinically suspected cases that tested negative for A. cantonensis-an-tibodies (n = 5) and patients with cerebral gnathostomiasis (n = 2) and neurocysticercosis (n = 2) were examined by a single-step polymerase chain reaction (PCR) method using the AC primers for the 66-kDa native protein gene. The PCR method detected A. cantonensis DNA in CSF samples from four of 10 serologically confirmed angiostrongyliasis cases. The PCR results were negative for the remaining CSF samples. The nucleotide sequences of three positive CSF-PCR samples shared 98.8-99.2% similarity with the reference sequence of A. cantonensis. These results indicate the potential application of this PCR assay with clinical CSF samples for additional support in the confirmation of eosinophilic meningitis due to A. cantonensis.     

The latex agglutination test versus counterimmunoelectrophoresis for rapid diagnosis of bacterial meningitis

A modified latex agglutination (LA) test was compared with Gram-staining and counterimmunoelectrophoresis (CIE) for the rapid detection in the cerebrospinal fluid (CSF) of antigen to Haemophilus influenzae type b, Neisseria meningitidis groups A, B and C, Escherichia coli K1, Streptococcus pneumoniae and group B streptococci, seven frequent causes of bacterial meningitis in children. Of 50 CSF samples from patients with culture-proven bacterial meningitis 90% were correctly shown by the LA test to contain antigen of the responsible organism. Gram-staining revealed organisms in 80% of 45 of these samples. In 75% of the 40 samples that were of sufficient volume for CIE, positive results for the appropriate antigen were obtained. The concentration of antigen detected in the CSF by the LA test varied from undetectable to 800 000 ng/ml. Patients with a high concentration (more than 2000 ng/ml or a positive result at dilutions of CSF over 1/8) were significantly more likely to have a poor response to therapy (two died and two had persistent pleocytosis or bacteria in the CSF) than patients with a lower concentration (4/16 v. 0/18, P < 0.05). After appropriate therapy was begun the concentration of antigen fell dramatically, but measurable amounts of antigen persisted in the CSF for up to 6 days. The LA test detected bacterial antigen at concentrations 2 to 70 times below the lower limit detected by CIE. In seven additional patients who had received antibiotics before lumbar puncture was performed the LA test detected antigen from meningitis-causing bacteria even though cultures of the CSF were sterile. In another 145 patients who did not have meningitis the results of the LA test were negative. The LA test, done as described in this article, is easier to perform than CIE and should be a useful addition to the diagnostic tests carried out on the CSF of any patient suspected of having meningitis.     

Conserved polymerase chain reaction primers fail in diagnosis of parasitic infections

We demonstrate that a set of previously described polymerase chain reaction primers used for detection of hemogregarines in reptiles will also amplify the same region of the 18S rRNA gene of reptiles, amphibians, mammals, and insects and thus should not be used for molecular diagnosis. These same primers have also been used to differentiate 2 species of Plasmodium that infect lizards. We provide evidence that the observed variance may have been dependent on parasitemia and not representative of actual molecular differences between the 2 parasite species.     

Specificity of immunoblotting analyses in eosinophilic meningitis

Angiostrongylus cantonensis and Gnathostoma spinigerum are the two most common causative parasites of eosinophilic meningitis (EOM). Serological tests are helpful tools for confirming the identity of the pathogen. Recent reports determined the specificity of such tests by using normal healthy controls. There have been limited studies done to rule out the cross-reactivity between these two causative parasites of EOM. This study aims to assess the specificity of the serological test in EOM by using each condition as a control for the other. Thirty-three patients with a diagnosis of EOM were enrolled. Sera from 22 patients with a positive 29-kDa antigenic diagnostic band of A. cantonensis were tested for the 21 and 24-kDa antigenic bands of G. spinigerum. Similarly, sera of 11 gnathostomiasis patients were tested for the 29-kDa diagnostic band for A. cantonensis. Only one patient in the angiostrongyliasis group had a positive result for the 21 and 24-kDa antigenic bands of G. spinigerum, while no gnathostomiasis patients showed a positive result for the 29-kDa antigenic band of A. cantonensis. The specificity of the 21 and 24-kDa antigenic bands for gnathostomiasis and the 29-kDa antigenic band for A. cantonensis was 95.5% and 100%, respectively. The antigenic bands for the diagnosis of gnathostomiasis and angiostrongyliasis in EOM were highly specific.     

SCID mice as models for parasitic infections

Mice with severe combined immunodeficiency (SCID mice) have become a favored model system for the study of many parasitic diseases. In this review, Samuel Stanley Jr and Herbert Virgin IV provide a brief overview of the biology of the SCID mouse, and review some examples of how the SCID mouse model has been applied to the study of the immunology of a number of different parasitic diseases.     

Use of the dot-immunogold assay for the rapid diagnosis of acute enteric infections

A simple method based on an immunodot assay using colloidal gold labels is proposed for the rapid diagnosis of a range of acute enteric infections. Owing to its rapidity, high sensitivity, and specificity, the method can be recommended for routine use in the laboratory diagnosis of enteric infections.     

The serodiagnosis of parasitic infections

Recently, the term of clinical immunoparasitology has been coined to indicate the application of immunological methods to the laboratory diagnosis of parasitic infections. In particular, serological diagnosis (indirect diagnosis) is useful especially in the cases of toxocarosis, trichinellosis, echinococcosis, cysticercosis, toxoplasmosis, amoebic abscess, some filariasis, visceral leishmaniasis, schistosomiasis. When possible, for infections caused by protozoa or helminths, the "gold standard" is represented by direct diagnosis performed by microscopic and/or macroscopic observation of the parasite. In any case, immunological results must be interpreted in consideration of the clinical picture of the patient and confirmed possibly by finding the parasite or its genome, even using molecular methods. Furthermore, since the presence of specific antibodies can reveal an acquired infection, but not necessarily a disease, it is particularly helpful, in addition to a qualitative evaluation, a quantitative one, by determining the serum antibody titre. After recovery, the antibody levels decrease, however, they may persist for long periods, for this reason they do not help in evaluating the treatment outcome. Interpretation of serological results may be difficult when the patients originate from areas where the suspected infection is endemic, in that case, a serum positivity could reflect an old exposition to the parasite, therefore it is not related to the present clinical status. Furthermore, serology may frequently result falsely negative in not immunocompetent subjects (organ transplanted, HIV positive individuals, premature babies, diabetics). Clinicians can interpret correctly the serological results only if the Parasitology laboratory inform them about the significant diagnostic values, the sensitivity and the specificity of the test in use. At present time, many diagnostic kits for immunoparasitology are commercially available, and industries are developing newer and newer ones (which are not always validated). In relation to this aspect, it should be helpful, for each of parasitic infection, to establish reference centers, not only to control the quality of commercial kits, but also as a reference point to those laboratories which use "in house" kits. To this regard, the recent establishment of a European Centre for Control of Infectious Diseases will help. The antigen characteristics (crude, E/S, recombinant, synthetic) for assays searching for antibodies (IHA, IFA, EIA, WB) of different classes, the controls to choose for these assays, the specimen requirements will be discussed. The recent findings on the serological diagnosis of intestinal protozoa infections, malaria, leishmaniasis, echinococcosis, cysticercosis, trichinellosis, toxocariasis, schistosomiasis, strongyloidiasis will be presented.