scholarly journals Immuno-PCR for Detection of Antigen to Angiostrongylus cantonensis Circulating Fifth-Stage Worms

2004 ◽  
Vol 50 (1) ◽  
pp. 51-57 ◽  
Author(s):  
Soi-Moi Chye ◽  
Shiu-Ru Lin ◽  
Ya-Lei Chen ◽  
Lee-Yi Chung ◽  
Chuan-Min Yen
Keyword(s):  
Monoclonal Antibody ◽  
Cerebrospinal Fluid ◽  
Parasitic Nematode ◽  
Pcr Amplification ◽  
Definitive Diagnosis ◽  
Angiostrongylus Cantonensis ◽  
Promising Technique ◽  
Serum Samples ◽  
Wilcoxon Rank Sum Test ◽  
Circulating Antigens

Abstract Background: Definitive diagnosis of infestation with Angiostrongylus cantonensis is difficult because the parasitic nematode is undetectable in the cerebrospinal fluid (CSF) of one-half of afflicted patients and the diagnostic sensitivity of ELISA for circulating worm antigens in patient sera is low. We studied immuno-PCR as a diagnostic tool. Methods: We studied 30 controls and 60 afflicted patients (30 confirmed by parasitologic analysis of CSF). We used a monoclonal antibody to capture circulating A. cantonensis antigens in serum samples. A DNA label generated by PCR amplification with biotinylated primer was bound by use of streptavidin to a biotinylated third antibody. Circulating antigens sandwiched by monoclonal antibody were detected by PCR amplification of the DNA label. Results: The detection limit of the ELISA was 100–1000 times higher than that of the immuno-PCR. The concentrations of circulating antigens in patients were markedly higher than those in controls (Wilcoxon rank-sum test, P <0.001). At a cutoff of 0.1 ng/L, sensitivity and specificity for immunodiagnosis of patients with angiostrongyliasis by immuno-PCR were 98% (95% confidence interval, 91–99%) and 100% (93–100%), respectively. The test was positive in all parasitologically confirmed cases. Conclusions: Immuno-PCR is a promising technique for diagnosis of A. cantonensis infestation.

scholarly journals sICAM-1 in meningoencephalitis due to Angiostrongylus cantonensis

2006 ◽  
Vol 64 (3a) ◽  
pp. 589-591 ◽  
Author(s):  
Alberto Juan Dorta-Contreras ◽  
Piotr Lewczuc ◽  
Elena Noris-García ◽  
María Teresa Interián-Morales ◽  
María Esther Magraner Tarrau ◽  
...  
Keyword(s):  
Immune Response ◽  
Cerebrospinal Fluid ◽  
Total Protein ◽  
Serum Analysis ◽  
Angiostrongylus Cantonensis ◽  
Brain Response ◽  
Serum Samples ◽  
Mean Values ◽  
Diagnostic Puncture ◽  
Eosinophilic Meningoencephalitis

INTRODUCTION: Angiostrongylus cantonensis meningoencephalitis is an emergent disease in the Americas. METHOD: Twelve children suffering from eosinophilic meningoencephalitis due to this parasite aged between 6-10 years were studied. Cerebrospinal fluid (CSF) and serum samples were taken simultaneously in the first diagnostic puncture at admission. RESULTS: All cases showed typical findings on the routine CSF and serum analysis: increased CSF total protein, increased Q (CSF/serum) albumin accompanied by eosinophilia in CSF. No intrathecal synthesis of immunoglobulins was found. Mean serum and CSF sICAM-1 values were 337.4 and 3.97 ng/mL. Qalbumin and QsICAM-1 mean values were 4.1 and 6.2 respectively. In 50% of the patients an increased brain-derived fraction of sICAM-1 was found. CONCLUSION: It may be suggested that a dynamic of the sICAM-1 brain derived fraction is perhaps associated to the immune response in the evolution of the disease.sICAM-1 may be an agent in negative feedback for eosinophils passage through the blood-CSF barrier into the inflammatory brain response.

High throughput sequencing of the Angiostrongylus cantonensis genome: a parasite spreading worldwide

Parasitology ◽  
2013 ◽  
Vol 140 (10) ◽  
pp. 1304-1309 ◽  
Author(s):  
ALESSANDRA L. MORASSUTTI ◽  
ANDREY PERELYGIN ◽  
MARCOS O. DE CARVALHO ◽  
LEANDRO NASCIMENTO LEMOS ◽  
PAULO MARCOS PINTO ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Parasitic Nematode ◽  
Definitive Diagnosis ◽  
Amino Acid Sequences ◽  
Angiostrongylus Cantonensis ◽  
Putative Orfs ◽  
Protein Targets ◽  
Antigenic Protein ◽  
Molecular Information

SUMMARYAngiostrongylus cantonensis is a parasitic nematode of rodents and a leading aetiological agent of eosinophilic meningitis in humans. Definitive diagnosis is difficult, often relying on immunodiagnostic methods which utilize crude antigens. New immunodiagnostic methods based on recombinant proteins are being developed, and ideally these methods would be made available worldwide. Identification of diagnostic targets, as well as studies on the biology of the parasite, are limited by a lack of molecular information on Angiostrongylus spp. available in databases. In this study we present data collected from DNA random high-throughput sequencing together with proteomic analyses and a cDNA walking methodology to identify and obtain the nucleotide or amino acid sequences of unknown immunoreactive proteins. 28 080 putative ORFs were obtained, of which 3371 had homology to other deposited protein sequences. Using the A. cantonensis genomic sequences, 156 putative ORFs, matching peptide sequences obtained from previous proteomic studies, were considered novel, with no homology to existing sequences. Full-length coding sequences of eight antigenic target proteins were obtained. In this study we generated not only the complete nucleotide sequences of the antigenic protein targets but also a large amount of genomic data which may help facilitate future genomic, proteomic, transcriptomic or metabolomic studies on Angiostrongylus.

scholarly journals Intrathecal Synthesis of Immunoglobulins in Eosinophilic Meningoencephalitis Due to Angiostrongylus cantonensis

1998 ◽  
Vol 5 (4) ◽  
pp. 452-455 ◽  
Author(s):  
Alberto Juan Dorta-Contreras ◽  
Hansotto Reiber
Keyword(s):  
Cerebrospinal Fluid ◽  
Pediatric Patients ◽  
Total Cell ◽  
Angiostrongylus Cantonensis ◽  
Total Cell Count ◽  
Barrier Dysfunction ◽  
Serum Samples ◽  
Clinical Recovery ◽  
Diagnostic Lumbar Puncture ◽  
Eosinophilic Meningoencephalitis

ABSTRACT Eosinophilic meningoencephalitis due to the nematodeAngiostrongylus cantonensis, which is endemic to Cuba, occurs in children and is due to accidental contact with soil snails. The course is less often fatal than in adult patients in southeastern Asia. Cerebrospinal fluid (CSF) and serum samples from 24 pediatric patients were analyzed and evaluated in CSF/serum quotient diagrams (Reiber graphs) to characterize the neuroimmunological response and the blood-CSF barrier dysfunction that occur in the course of the disease. At the time of the first diagnostic lumbar puncture, together with eosinophilic pleocytosis (1,920 ± 400 cells/μl), intermediate blood-CSF barrier dysfunction (i.e., an increased CSF/serum albumin quotient) with no intrathecal immunoglobulin G (IgG), IgA, and IgM class response was observed in all cases. Seven days later, at the time of early clinical recovery, the blood-CSF barrier dysfunction was normalized in 75% of the patients, but meanwhile, intrathecal immunoglobulin synthesis emerged in all cases, as either a two-class response (IgG and IgA in 85% of the patients) or a three-class response (IgG, IgA, and IgM; 30%). The fraction of eosinophilic cells (40%) remained large despite a decreasing total cell count. The neuroimmunological pattern of this inflammatory response to the parasite and its toxins is discussed with regard to the CSF patterns of other infectious diseases caused by bacteria or viruses.

Abstract T P351: Unbound Iron and Ceruloplasmin Levels in Cerebrospinal Fluid Are Associated with Development of Deep Cerebral Infarcts Following Subarachnoid Hemorrhage

Stroke ◽  
2014 ◽  
Vol 45 (suppl_1) ◽  
Author(s):  
Lauren Koffman ◽  
Gabor Toth ◽  
M. Shazam Hussain ◽  
Magdy Selim ◽  
Peter Rasmussen ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Subarachnoid Hemorrhage ◽  
Cerebral Infarcts ◽  
Active Iron ◽  
Redox Active ◽  
Wilcoxon Rank Sum Test ◽  
Radiological Data ◽  
Experimental Subarachnoid Hemorrhage ◽  
Preliminary Study ◽  
Iron Redox

Introduction: Iron-dependent formation of reactive oxygen species has been implicated in the development of vasospasm (VSP) and neuronal injury following experimental subarachnoid hemorrhage (SAH). We report the association between unbound (“free”) iron in CSF of SAH patients and the risk of angiographic vasospasm and cerebral infarcts (CI) on neuroimaging from a recently completed pilot study. Methods: Samples of cerebrospinal fluid (CSF) were obtained on days 1, 3, and 5. A fluorometric assay that relies on an oxidation sensitive probe was used to measure redox active iron (REDOX-Fe). Ceruloplasmin (Cp) concentration and levels of malondialdehyde (MDA), a marker of lipid peroxidation were also measured. We prospectively collected and recorded demographic, clinical, and radiological data. Logistic regression and Wilcoxon Rank Sum test were used. Results: Five of 12 patients developed angiographic VSP (41.6%) and eight developed CI (66.6%). Mean REDOX-Fe was higher in patients with CI (3.96 ± 0.97 Vs. 2.77 ± 0.87 mcg/dl, p 0.07), particularly in patients with deep-seated strokes (4.56 ± 0.67 Vs. 3.35 ± 0.89, p 0.03). Levels of Cp at day 3 were lower in patients with deep strokes (34,092 ± 23,780 Vs. 86,045 ± 34,752 ng/ml, p 0.03). A trend towards higher REDOX-Fe on day 3 in patients who developed VSP (4.52 ± 1.16 Vs. 2.96 ± 0.71, p 0.07), and lower Cp levels on day 5 (45,033 ± 29,079 Vs. 63,044 ± 24,821, p 0.1) was found. Levels of MDA were higher in patients who developed CI (10.36 ± 4.36 Vs. 5.9 ± 4.2 nmol, p 0.08). Conclusions: In this preliminary study we found higher concentrations of redox active iron in CSF of SAH patients who develop deep-seated CI on neuroimaging. Evidence of increased oxidative damage correlated with development of CI. A possible association between non-protein bound iron and angiographic VSP is suggested as well. Ceruloplasmin may exert a protective effect in this setting. Further studies are needed to validate these findings.

scholarly journals Association Between Vancomycin Blood Brain Barrier Penetration and Clinical Response in Postsurgical Meningitis

2017 ◽  
Vol 20 ◽  
pp. 161 ◽  
Author(s):  
Qing Wang ◽  
Si Chen ◽  
Yan-Gang Zhou ◽  
Ping Xu ◽  
Yi-Ping Liu ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Clinical Response ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Serum Samples ◽  
Cell Counts ◽  
Blood Cell Counts ◽  
Blood Brain ◽  
Barrier Penetration ◽  
White Blood Cell Counts

PURPOSE: This study investigated the association between vancomycin blood brain barrier penetration and clinical response in patients with postsurgical meningitis. METHODS: Adult patients with postsurgical meningitis were recruited. Eligible patients received vancomycin 500 mg every 6 h for at least 5 days. On day 3 or 4, cerebrospinal fluid (CSF) and simultaneous serum samples were obtained to determine CSF minimum concentrations (Cmin), serum Cmin and CSF to serum Cmin ratio. RESULTS: Twenty-two patients (14 men and 8 women; mean age of 52.6± 12.1 years) were recruited. The vancomycin Cmin was 3.63 ± 1.64 mg/L in CSF and 13.38 ± 5.36 mg/L in serum, with the CSF to serum Cmin ratio of 0.291 ± 0.118. The Cmin in serum and in CSF showed a significant correlation (p=0.005, r =0.575). The vancomycin CSF Cmin had a significant correlation with the decline of white blood cell counts (WBCs) in CSF (p=0.003, r =0.609). CSF Cmin, serum Cmin and CSF to serum Cmin ratio all showed no significant correlation with clinical response (p=0.335, 0.100, 0.679, respectively). CONCLUSIONS: There was a positive correlation between serum Cmin and CSF Cmin. However, only CSF Cmin is positively correlated with WBCs improvement in CSF. All other parameters such as serum Cmin, CSF Cmin and CSF to serum Cmin ratio had no correlation with clinical response. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

Osmotic Blood-Brain Barrier Modification: Monoclonal Antibody, Albumin, and Methotrexate Delivery to Cerebrospinal Fluid and Brain

Neurosurgery ◽  
1985 ◽  
Vol 17 (3) ◽  
pp. 419-423 ◽  
Author(s):  
Edward A. Neuwelt ◽  
Peggy A. Barnett ◽  
Christopher I. McCormick ◽  
Eugene P. Frenkel ◽  
John D. Minna
Keyword(s):  
Monoclonal Antibody ◽  
Cerebrospinal Fluid ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Blood Brain

scholarly journals Detection of circulating antigens in serum samples of mice experimentally infected with Trichinella spiralis by a sandwich ELISA based on IgY

2014 ◽  
Vol 51 (3) ◽  
pp. 181-189 ◽  
Author(s):  
F. Jing ◽  
J. Cui ◽  
R. Liu ◽  
L. Liu ◽  
P. Jiang ◽  
...  
Keyword(s):  
Post Infection ◽  
Trichinella Spiralis ◽  
Sandwich Elisa ◽  
Egg Yolk ◽  
Mouse Serum ◽  
Serum Samples ◽  
Egg Yolk Immunoglobulin ◽  
Muscle Larvae ◽  
Sandwich Elisa Assay ◽  
Circulating Antigens

AbstractIn the present study, a sandwich ELISA based on IgY (egg yolk immunoglobulin) was developed for detection of circulating antigens (CAg) in sere of mice experimentally infected with Trichinella spiralis. The IgY-sandwich ELISA assay involved the use of chicken antibody IgY against excretory-secretory (ES) antigens of Trichinella spiralis muscle larvae as a capture antibody and mouse polyclonal antibody IgG to ES antigens as a detecting antibody. This method was able to detect as little as 3 ng/ml of ES antigens added to normal mouse serum. A group of sixteen mice was orally inoculated with 500 T. spiralis muscle larvae per animal. The serum samples from the infected mice were taken during 1–35 days post-infection (dpi). The CAg was detectable as early as 8 dpi in the sera of infected mice. The level of CAg increased dramatically during 13–15 dpi and reached a peak at 22 dpi and remained a plateau for 3 days, then declined gradually. Another peak of CAg occurred at 31 dpi. The anti-Trichinella antibodies was first detected in 14.3 % of the infected mice at 2 weeks post-infection (wpi), and reached a peak positive rate of 100 % at 5 wpi. Moreover, the infected mice were treated with abendazole at 5 wpi and the serum CAg levels increased significantly during 2–6 days posttreatment (dpt) and then declined rapidly during 8–14 dpt. By 42 dpt, the CAg levels decreased to the undetected level, but the detection rate of antibodies was still 100 %. The IgY-sandwich ELISA appears to be a sensitive for detection of antigenemia of T. spiralis and valuable to judge the efficacy of chemotherapy in trichinellosis.

scholarly journals Oligoclonal IgG bands in the cerebrospinal fluid of portuguese patients with multiple sclerosis: negative results indicate benign disease

2005 ◽  
Vol 63 (2b) ◽  
pp. 375-379 ◽  
Author(s):  
Maria José Sá ◽  
Lucinda Sequeira ◽  
Maria Edite Rio ◽  
Edward J. Thompson
Keyword(s):  
Multiple Sclerosis ◽  
Cerebrospinal Fluid ◽  
Benign Disease ◽  
Serum Samples ◽  
Negative Results ◽  
Significant Difference ◽  
Relapsing Remitting ◽  
The Mean ◽  
Oligoclonal Igg ◽  
Oligoclonal Igg Bands

We assessed the frequency of cerebrospinal fluid (CSF) restricted oligoclonal IgG bands (IgG-OCB) in Portuguese multiple sclerosis (MS) patients and its relationship with outcome. Paired CSF/serum samples of 406 patients with neurological disorders were submitted to isoelectric focusing with immunodetection of IgG. Ninety-two patients had definite MS; non-MS cases were assembled in groups inflammatory/infectious diseases (ID, n=141) and other/controls (OD, n=173). We found in the MS group: mean duration, 38.9 months; clinically isolated syndromes, 24%; relapsing/remitting course (RR), 65%; in RR patients the mean EDSS was 2.1 and the mean index of progression was 0.31. Positive patterns significantly predominated in MS (82.6%; ID, 40.4%; OD, 3.5%). The sensitivity and the specificity of positive IgG-OCB for MS diagnosis was 82.6% and 79.9%, respectively. The sole statistically significant difference in the MS group was the lower progression index observed in negative cases. We conclude that the frequency of positive IgG-OCB patterns in our MS patients fits most values reported in the literature, and that negative results indicate benign disease.

scholarly journals Serum and cerebrospinal fluid host proteins indicate stroke in children with tuberculous meningitis

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250944
Author(s):  
Charles M. Manyelo ◽  
Novel N. Chegou ◽  
James A. Seddon ◽  
Candice I. Snyders ◽  
Hygon Mutavhatsindi ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Tuberculous Meningitis ◽  
Multiple Testing ◽  
Serum Proteins ◽  
Lipocalin 2 ◽  
Host Proteins ◽  
Serum Samples ◽  
Csf Proteins ◽  
Study Participants

Introduction Stroke is a common complication in children with tuberculous meningitis (TBM). Host proteins may give us insight into the mechanisms of stroke in TBM and serve as biomarkers for detection of stroke, however, they have not been widely explored. In this study, we compared the concentrations of cerebrospinal fluid (CSF) and serum proteins between children who had TBM-related stroke and children with TBM without stroke. Methods We collected CSF and serum from 47 children consecutively admitted to the Tygerberg Academic Hospital in Cape Town, South Africa between November 2016, and November 2017, on suspicion of having TBM. A multiplex platform was used to measure the concentrations of 69 host proteins in CSF and serum from all study participants. Results After classification of study participants, 23 (48.9%) out of the 47 study participants were diagnosed with TBM, of which 14 (60.9%) demonstrated radiological arterial ischemic infarction. The levels of lipocalin-2, sRAGE, IP-10/ CXCL10, sVCAM-1, MMP-1, and PDGF-AA in CSF samples and the levels of D-dimer, ADAMTS13, SAA, ferritin, MCP-1/ CCL2, GDF-15 and IL-13 in serum samples were statistically different between children who had TBM-related stroke and children with TBM without stroke. After correcting for multiple testing, only the levels of sVCAM-1, MMP-1, sRAGE, and IP-10/ CXCL10 in CSF were statistically different between the two groups. CSF and serum protein biosignatures indicated stroke in children diagnosed with TBM with up to 100% sensitivity and 88.9% specificity. Conclusion Serum and CSF proteins may serve as biomarkers for identifying individuals with stroke amongst children diagnosed with TBM at admission and may guide us to understand the biology of stroke in TBM. This was a pilot study, and thus further investigations in larger studies are needed.

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