Cerebral Spinal Fluid Specimens Research Articles

Novel intertypic recombination of Echovirus 11 in the Enterovirus species B.

Enteroviruses (EVs), single-stranded, positive-sense RNA viruses, can be classified into four species (A-D), which have previously been linked to a diverse range of disease manifestations and infections affecting the central nervous system. In the Enterovirus species B (EV-B), Echovirus type 11 (E11) has been observed to occasionally circulate in Taiwan, which was responsible for an epidemic of enterovirus infections in 2018. Here, 48 clinical specimens isolated in 2003, 2004, 2009, and 2018 were collected for the high-throughput sequencing. Notably, we identified 2018 Taiwanese strains having potential recombinations in the 3D gene, as well as one 2003 strain having a double recombination with E6 and Coxsackievirus B5 in the P2 and P3 regions, respectively. Additionally, one amino acid signature mutated from the Histidine (H) in throat swab specimens to the Tyrosine (Y) in cerebral spinal fluid specimens was detected at position 1496 (or 57) of the genomic coordinate (or 3A gene) to further demonstrate intra-host evolution in different organs. In conclusion, this study identifies potential intertypic recombination events and an intra-host signature mutation in E11 strains, isolated during a 2018 neurological disease outbreak in Taiwan, contributing to our understanding of its evolution and pathogenesis.

CSF cytology of common primary CNS neoplasms categorized by CNS WHO 2021.

The detection of neoplastic cells in cerebral spinal fluid (CSF) is pivotal for the management of patients with central nervous system (CNS) tumours. This article delves into the CSF cytological characteristics of common CNS neoplasms, aligning with the 2021 World Health Organization (WHO) classification of CNS tumours. A retrospective review of CSF specimens positive for primary CNS neoplasms was performed at three tertiary medical centres. Only cases that had histopathologic confirmation and/or molecular workup were included. Common primary CNS neoplasms seen in CSF cytology specimens include medulloblastoma, (non-WNT/non-SHH as well as SHH-activated and TP53 mutant), pineoblastoma, atypical teratoid/rhabdoid tumour (AT/RT), IDH-wildtype glioblastoma, and primary diffuse large B-cell lymphoma of the CNS. Ependymomas and germinomas can also have CSF involvement but are less common. Although the typical histologic architecture of these tumours may not be preserved in the CSF, unique cytomorphologic features such as nuclear moulding, nuclear pleomorphism, rhabdoid cells, prominent nucleoli and rosette formation can still be appreciated. Adopting the updated terminology and correlating cytologic observations with molecular findings will streamline the diagnostic process, reducing the complexities and ambiguities pathologists often encounter when analysing CSF specimens for potential primary CNS neoplasms.

Clinicopathologic and Genomic Landscape of Non-Small Cell Lung Cancer Brain Metastases.

BackgroundIn patients with non-small cell lung cancer (NSCLC), 10%-40% will eventually develop brain metastases. We present the clinicopathologic, genomic, and biomarker landscape of a large cohort of NSCLC brain metastases (NSCLC-BM) samples.Materials and MethodsWe retrospectively analyzed 3035 NSCLC-BM tested with comprehensive genomic profiling (CGP) during routine clinical care. In addition, we compared the NSCLC-BM to a separate cohort of 7277 primary NSCLC (pNSCLC) specimens. Finally, we present data on 67 paired patients with NSCLC-BM and pNSCLC.ResultsComprehensive genomic profiling analysis of the 3035 NSCLC-BMs found that the most frequent genomic alterations (GAs) were in the TP53, KRAS, CDKN2A, STK11, CDKN2B, EGFR, NKX2-1, RB1, MYC, and KEAP1 genes. In the NSCLC-BM cohort, there were significantly higher rates of several targetable GAs compared with pNSCLC, including ALK fusions, KRAS G12C mutations, and MET amplifications; and decreased frequency of MET exon14 skipping mutations (all P < .05). In the subset of NSCLC-BM (n = 1063) where concurrent PD-L1 immunohistochemistry (IHC) was performed, 54.7% of the patients with NSCLC-BM were eligible for pembrolizumab based on PD-L1 IHC (TPS ≥ 1), and 56.9% were eligible for pembrolizumab based on TMB-High status. In addition, in a series 67 paired pNSCLC and NSCLC-BM samples, 85.1% (57/67) had at least one additional GA discovered in the NSCLC-BM sample when compared with the pNSCLC sample.ConclusionsHerein, we defined the clinicopathologic, genomic, and biomarker landscape of a large cohort of patients with NSCLC-BM which can help inform study design of future clinical studies for patients with NSCLC with BM. In certain clinical situations, metastatic NSCLC brain tissue or cerebral spinal fluid specimens may be needed to fully optimize personalized treatment.

A Practical Strategy of Monitoring Minimal Residue Disease and Intervention for Central Nervous System Relapse of Childhood Acute Lymphoblastic Leukemia

To investigate monitoring minimal residual disease (MRD) using cerebral spinal fluid for predicting central nervous system leukemia (CNSL) and treatment. There is no survival difference between enhanced triple intrathecal therapy (ETIT) and cranial radiation for CNSL patients with positive morphology and MRD. Positive MRD correlated with CNSL, whereas negative MRD indicated a lower chance of CNSL recurrence. Altogether 79 cerebral spinal fluid specimens indicating negative morphology but positive MRD were given either ETIT or conventional triple intrathecal therapy. The ETIT group indicated lower relapse. Flow cytometry is sensitive to predict CNSL and ETIT is a potent intervention.

Predictors of Bacterial Meningitis in Resource-Limited Contexts: An Angolan Case

BackgroundDespite the great morbidity and mortality that childhood bacterial meningitis (BM) is experiencing in Africa, diagnosis of BM in resource-limited contexts is still a challenge. Several algorithms and clinical predictors have been proposed to help physicians in decision-making but a lot of these markers used variables that are calculable only in well-equipped laboratories. Predictors or algorithm based on parameters that can be easily performed in basic laboratories can help significantly in BM diagnosis, even in resource-limited settings, rural hospitals or health centers.ResultsThis retrospective study examined 145 cerebral-spinal fluid (CSF) specimens from children from 2 months to 14 years. CSF specimens were divided into two groups, according to the presence or not of a clinical diagnosis of BM. For each specimen, CSF aspect, CSF white blood cells (WBC) count, CSF glucose and protein concentration were analyzed and statistical analysis were performed. CSF WBC count ≥10/µl is no more a valuable predictor of BM. CSF protein concentration ≥50 mg/dl has a better sensitivity for BM diagnosis and when used with CSF glucose concentration ≤40 mg/dl, can help to diagnose correctly almost all the BM cases. An algorithm including CSF protein concentration, glucose concentration and WBC count has been proposed to rule out BM and to correctly diagnose it.ConclusionsIn resource-limited health centers, the availability of a combination of easy-to-obtain parameters can significantly help physicians in BM diagnosis. The prompt identification of a BM case can be rapid treated or transferred to adequate structures and can modify the outcome in the patient.

Evaluation of the Diagnostic Utility of Cerebral Spinal Fluid (CSF) Flow Cytometry (FC) In Detection of Central Nervous System (CNS) Involvement by Hematological Malignancy.

AbstractAbstract 3837 Introduction:Flow cytometry (FC) is more sensitive but more expensive than cytomorphology in detection central nervous system (CNS) involvement by hematological malignancy. Previous studies have shown that cerebral spinal fluid (CSF) pleocytosis is absent in about one third of patients with positive FC findings. FC is often performed to investigate CNS involvement on all CSF specimens regardless of cell counts or morphology, which may lead over utilization of FC. Here we investigate the diagnostic value of FC evaluation of CSF specimens with low white blood cell (WBC) count. Methods:After IRB approval, we retrospectively reviewed the FC results of CSF samples studied for CNS involvement by hematological malignancy between Jul 2000 and Jun 2010 at William Beaumont Hospital, MI. The FC results were correlated with CSF WBC count, cytomorphology, and documented leptomeningeal symptoms at the time of the sample obtained. Results:Of 421 cases included in this study, 73 were acute leukemia and 348 were lymphoma. The mean age was 59 (range from 9–90). 9.5% (40/421) of patients had leptomeningeal symptoms. 13.3% (56/421) had atypical CSF WBC cytomorphology. 11% (46/421) had positive FC. Patients with either atypical cytomorphology or leptomengeal symptoms were more likely have positive FC (p<0.001). Of the 46 patients with positive FC, 32.6% (15/46) of patients did not have CSF leptomengeal symptoms and 19.5% (9/46) had neither CSF pleocytosis (<10/μ l) nor atypical cytomorphology. We did not detect lymphoma or leukemia cells by FC in CSF samples with WBC <3/μ l, which represented 74.7% (314/421) of the specimens studied. Conclusion:In this study, we found that FC is more sensitive and specific than leptomeningeal symptoms or cytomorphology in detection of CNS involvement by hematological malignancy in both acute leukemia and lymphoma. However, its sensitivity is dependent on numbers of cells potentially available for analysis. Flow cytometry may not be indicated when CSF WBC count is extremely low (<3/μ l) which is invariably predictive of negative results. Disclosures:No relevant conflicts of interest to declare.

Evaluation of Three Commercially Available Japanese Encephalitis Virus IgM Enzyme-Linked Immunosorbent Assays

We evaluated performance of three commercial Japanese encephalitis virus (JEV) IgM antibody capture enzyme-linked immunosorbent assay (MAC ELISA) kits with a panel of serological specimens collected during a surveillance project of acute encephalitis syndrome in India and acute meningitis and encephalitis syndrome in Bangladesh. The serum and cerebral spinal fluid specimens had been referred to the Centers for Disease Control and Prevention (CDC) for confirmatory testing. The CDC results and specimen classifications were considered the reference standard. All three commercial kits had high specificity (95-99.5%), but low sensitivities, ranging from 17-57%, with both serum and cerebrospinal fluid samples. Specific factors contributing to low sensitivity compared with the CDC ELISA could not be determined through further analysis of the limits and dilution end points of IgM detection.

Development of an Ultra-Rapid Diagnostic Method Based on Heart-Type Fatty Acid Binding Protein Levels in the CSF of CJD Patients

Creutzfeldt-Jakob disease (CJD) is a transmissible, fatal, neurodegenerative disease in humans. Recently, various drugs have been reported to be useful in the treatment of CJD; however, for such treatments to be useful it is essential to rapidly and accurately diagnose CJD. 124 CJD patients and 87 with other diseases causing rapid progressive dementia were examined. Cerebral spinal fluid (CSF) from CJD patients was analyzed by 2D-PAGE and the protein expression pattern was compared with that from healthy subjects. One of three CJD-specific spots was found to be fatty acid binding protein (FABP), and heart-type FABP (H-FABP) was analyzed as a new biochemical marker for CJD. H-FABP ELISA results were compared between CJD patients and patients with other diseases (n=211). Visual readout accuracy of the Rapicheck(®) H-FABP test panel for CSF was analyzed using an independent measure of CSF H-FABP concentration. The distribution of H-FABP in the brains of CJD patients was examined by immunohistochemistry. ELISA sensitivity and specificity were 90.3% and 92.9%, respectively, and Rapicheck(®) H-FABP sensitivity and specificity were 87.9% and 96.0%, respectively. ELISA and Rapicheck(®) H-FABP assays provided comparable results for 14-3-3 protein and total tau protein. Elevated H-FABP levels were associated with an accumulation of abnormal prion protein, astrocytic gliosis, and neuronal loss in the cerebral cortices of CJD patients. In conclusion, Rapicheck(®) H-FABP of CSF specimens enabled quick and frequent diagnosis of CJD. H-FABP represents a new biomarker for CJD distinct from 14-3-3 protein and total tau protein.

Distribution and antibiotic resistance of bacteria isolated from cerebral spinal fluid in postoperative patients with traumatic brain injury: a surveillance

Objective To monitor epidemiological distribution and antibiotic resistance of bacteria isolated from cerebral spinal fluid in neurosurgically treated patients. Methods Bacteria isolated from cerebral spinal fluid specimens and antibiotic resistance identification results of patients admitted into our department from January 2003 to December 2007 were collected to analyze distribution and antibiotic resistance of the pathogens. Results Of 437 isolates, Gram-positive bacilli and Gram-negative bacilli accounted for 73.5% (321 isolates) and 26.5% ( 116 isolates), respectively. The first five most fre-quently isolated pathogens were staphylococcus epidermidis (55.6%), staphylococcus aurens (15. 3% ), enterobacter (6.6%), aeinetobacter (5.3%) and pseudomonas aeruginosa (3.0%). The anti-bacterials with highest susceptibility to Gram-negative bacilli were imipenem ( 83.9% ), amikacin ( 68. 8% ), ciprofloxacin and tobramycin (67.4%), cefepime (61.3%) and ceftazidime (69.2%). The other antibacterials had susceptibility of less than 60%. Conclusions The prevailing pathogens in post-operative intracranial infection are Gram-pnsitive bacilli, especially staphylococcus epidermidis and staphylococcus anreu. Data collected in present study may provide valuable information for prophylactic and empirical antibiotic use in post-operative intracranial infection. Key words: Brain injuries; Cerebrospinal fluid; Antibiotics; Infection

Diagnosis of Infections Caused by Pathogenic Free-Living Amoebae

Naegleria fowleri, Acanthamoeba spp., Balamuthia mandrillaris, and Sappinia sp. are pathogenic free-living amoebae. N. fowleri causes Primary Amoebic Meningoencephalitis, a rapidly fatal disease of the central nervous system, while Acanthamoeba spp. and B. mandrillaris cause chronic granulomatous encephalitis. Acanthamoeba spp. also can cause cutaneous lesions and Amoebic Keratitis, a sight-threatening infection of the cornea that is associated with contact lens use or corneal trauma. Sappinia pedata has been identified as the cause of a nonlethal case of amoebic encephalitis. In view of the potential health consequences due to infection with these amoebae, rapid diagnosis is critical for early treatment. Microscopic examination and culture of biopsy specimens, cerebral spinal fluid (CSF), and corneal scrapings have been used in the clinical laboratory. For amoebic keratitis, confocal microscopy has been used to successfully identify amoebae in corneal tissue. More recently, conventional and real-time PCR assays have been developed that are sensitive and specific for the amoebae. In addition, multiplex PCR assays are available for the rapid identification of these pathogens in biopsy tissue, CSF, and corneal specimens.

Meningitis due toPrototheca wickerhamii: rare case in China

A rare case of Prototheca wickerhamii meningitis is reported in a patient without any underlying immunodeficient condition. Wet-mount microscopy and culture of cerebral spinal fluid specimens, along with temperature resistance, cycloheximide tolerance, carbohydrates assimilation, including API 20C AUX tests of the isolated etiologic agent were performed. In addition, transmission and scanning electron microscopy studies and in vitro antifungal susceptibility tests were conducted. Through the combination of these investigations, the isolate was identified as P. wickerhamii and the patient was successfully treated with intravenous amphotericin B and itraconazole. This is the first detailed report of meningitis caused by P. wickerhamii in China.

Performance of a Commercial Immunoglobulin M Antibody Capture Assay Using Analyte-Specific Reagents To Screen for Interfering Factors during a West Nile Virus Epidemic Season in Nebraska

In 2003, the Nebraska Public Health Laboratory tested more than 10,371 serum and 516 cerebral spinal fluid specimens. Results showed that without performing the interfering factors screen for specimens in the low positive index value range of >1.1 to <or=3.5, a false positivity rate of 6.5% would have been realized.

Isolation and detection of Borrelia burgdorferi DNA from cerebral spinal fluid, synovial fluid, blood, urine, and ticks using the Roche MagNA Pure system and real-time PCR

The Roche MagNA Pure automated nucleic acid extraction system was tested for its ability to extract Borrelia burgdorferi DNA from a diverse set of spiked specimen types including blood, cerebral spinal fluid, synovial fluid, urine and ticks. A method comparison between MagNA Pure automated extraction and manual extraction, using either QIAamp columns or phenol/chloroform extraction, showed equivalent detection sensitivities for all methodologies with all specimen types (except for urine, in which case QIAamp extraction was twofold less sensitive). Eighty positive clinical specimens (as determined by an independent testing method), including 76 synovial fluid, and 4 cerebral spinal fluid specimens, were found to be positive by the MagNA Pure/real-time PCR method of extraction and detection. This data shows that the MagNA Pure system can be used to extract B. burgdorferi DNA from clinical specimens, and when combined with real-time PCR, the result is an extremely sensitive assay with limited hands on time and rapid turn around times.

Cryptococcal meningoencephalitis in a patient with Hodgkin's lymphoma: successful treatment with fluconazole

The authors report a case of cryptococcal meningoencephalitis in a 22-year-old male with Hodgkin's disease at third stage B. Cryptococcus neoformans was isolated from cerebral spinal fluid (CSF) and blood specimens. Latex antigen agglutination tests and India ink preparations also were positive for this pathogenic yeast. Drug treatment for a week with 20 mg i.v./day of amphotericin B in combination with 2.5 g i.v./day of 5-fluorocytosine did not lead to any improvement in the patient. The patient became asymptomatic after the first week of treatment with a daily dose of 400 mg of fluconazole, administered intravenously. After two months of this regimen, all laboratory tests on CSF and blood specimens were negative.

Value and interpretation of serological tests for the diagnosis of cryptococcosis.

MAXIMAL SEROLOGICAL DIAGNOSIS OF CRYPTOCOCCOSIS MAY BE ACCOMPLISHED THROUGH THE CONCURRENT USE OF THREE TESTS: the latex agglutination (LA) test for cryptococcal antigen, and the indirect fluorescent antibody (IFA) and tube agglutination (TA) tests for Cryptococcus neoformans antibodies. These tests were applied to 141 serum and cerebral spinal fluid specimens from 66 culturally proven cases of cryptococcosis and to 42 sera from normal subjects and from patients with other systemic mycotic diseases. The LA test was sensitive and completely specific; of the sera from proven cases, 55% were positive. With the TA test, 37% of the specimens were positive and the test was highly specific. With the IFA test, 38% of the specimens were positive and the test appears to be the least specific of the three. Cross-reactions were most evident with blastomycosis and histoplasmosis case sera. When the three tests were used concurrently, 87% of the cryptococcosis case specimens were positive and permitted a presumptive diagnosis of C. neoformans infections in 61 (92%) of the 66 patients whose specimens were examined.

Value and Interpretation of Serological Tests for the Diagnosis of Cryptococcosis

MAXIMAL SEROLOGICAL DIAGNOSIS OF CRYPTOCOCCOSIS MAY BE ACCOMPLISHED THROUGH THE CONCURRENT USE OF THREE TESTS: the latex agglutination (LA) test for cryptococcal antigen, and the indirect fluorescent antibody (IFA) and tube agglutination (TA) tests for Cryptococcus neoformans antibodies. These tests were applied to 141 serum and cerebral spinal fluid specimens from 66 culturally proven cases of cryptococcosis and to 42 sera from normal subjects and from patients with other systemic mycotic diseases. The LA test was sensitive and completely specific; of the sera from proven cases, 55% were positive. With the TA test, 37% of the specimens were positive and the test was highly specific. With the IFA test, 38% of the specimens were positive and the test appears to be the least specific of the three. Cross-reactions were most evident with blastomycosis and histoplasmosis case sera. When the three tests were used concurrently, 87% of the cryptococcosis case specimens were positive and permitted a presumptive diagnosis of C. neoformans infections in 61 (92%) of the 66 patients whose specimens were examined.