High frequency of Taenia solium antigen positivity in patients admitted for neurological disorders in the Rural Hospital of Mosango, Democratic Republic of Congo.

BACKGROUND: The epidemiology of human cysticercosis and neurocysticercosis, caused by the larval stage of the pork tapeworm Taenia solium, is not well known in the Democratic Republic of Congo (DRC). Within a multicenter etiological and diagnostic study conducted by the NIDIAG consortium ("Better Diagnosis for Neglected Infections") and investigating several challenging syndromes, we consecutively evaluated from 2012 to 2015 all patients older than 5 years presenting with neurological disorders (neurology cohort) and with fever > 7 days (persistent fever cohort) at the rural hospital of Mosango, province of Kwilu, DRC. In both cohorts, etiological diagnosis relied on a systematic set of reference laboratory assays and on pre-established clinical case definitions. No neuroimaging was available in the study hospital. In this study, we determined the frequency of T. solium infection in both cohorts and explored in the neurology cohort its association with specific neurological presentations and final etiological diagnoses. METHODS: We conducted a post-hoc descriptive and analytic study on cysticercosis in the neurology and persistent fever cohorts, based on the presence in serum samples of circulating T. solium antigen using the B158/B60 enzyme-linked immunosorbent assay (ELISA) and of cysticercosis IgG using the LDBIO Cysticercosis Western Blot IgG assay. RESULTS: For the neurology cohort, 340 samples (of 351 enrolled patients) were available for analysis (males: 46.8%; mean age: 38.9 years). T. solium antigen positivity was found in 43 participants (12.6%; 95% confidence interval [CI] 9.3-16.7%), including 9 of 60 (15%) patients with epilepsy. Among the 148 samples available from the persistent fever cohort (males: 39.9%; mean age: 19.9 years), 7 were positive in the T. solium antigen ELISA (4.7%; 95% CI 1.9-9.5%; P = 0.009 when compared to the neurology cohort). No significant association was found within the neurology cohort between positivity and clinical presentation or final diagnoses. Of note, the IgG antibody-detecting assay was found positive in only four (1.3%) of the participants of the neurology cohort and in none of the persistent fever cohort. CONCLUSIONS: T. solium antigen positivity was found in at least 10% of patients admitted with neurological disorders in the Kwilu province, DRC, with no specific pattern of presentation. Further neuroimaging studies should be used to confirm whether neurocysticercosis is prevalent in this region.

Sorting out difficulties in immunological diagnosis of neurocysticercosis: Development and assessment of real time loop mediated isothermal amplification of cysticercal DNA in blood.

PURPOSE OF THE STUDY: Among various immunological tests available for diagnosis of neurocysticercosis (NCC), only EITB (Electroimmunotransfer blot for detection of anticysticercal antibodies) had gained widespread acceptance. However EITB is not available widely and is costly (Indian rupees 15,000/- approximately). We evaluated utility of Loop mediated isothermal amplification (LAMP) assay for detection of Taenia solium cox1 gene in blood of patients with NCC. PATIENTS AND METHODS: Current study included 100 consecutive patients of NCC at a tertiary teaching hospital in Northern India. All the patients underwent detailed history and examinations as well as gadolinium enhanced magnetic resonance imaging of brain. LAMP assay was performed in all the patients. The results were compared with 50 controls. RESULTS: LAMP detected Taenia Solium cox1 gene in 74% of all blood samples in patients of NCC.T he overall sensitivity of LAMP assay for detection of cox1 gene was 74% in all patients with NCC, 71.8% in patients with intraparenchymal brain cysts only and 86.7% of patients with extraparenchymal brain cysts with or without intraparenchymal brain cysts. The overall specificity of LAMP assay was 90% in all these three subgroups. The positive predictive value of real time LAMP assay was close to 93% for almost all forms of NCC- both solitary and multiple while negative predictive value ranged from 57 to 64%. CONCLUSION: Real time LAMP assay of blood for detection of Taenia solium cox1 gene appears to be a promising toll for diagnosis of NCC.

Neurocysticercosis serodiagnosis: mimotope-based synthetic peptide as potential biomarker.

Herein, we evaluate a mimotope-based synthetic peptidenamed NC41 to diagnose neurocysticercosis (NC), a neglected parasitic disease and a major cause of epilepsy worldwide. NC41 synthetic peptide was evaluated to diagnose NC, and total saline extract from Taenia solium metacestodes (SE) was used as control. Serum samples from patients with NC (n = 40), other parasitic diseases (n = 43), and healthy individuals (n = 40) were tested. Diagnostic parameters such as sensitivity (Se), specificity (Sp), likelihood ratio (LR), and area under curve (AUC) were calculated using receiver operating characteristic (ROC) curves. The sequence from T. solium phosphoenolpyruvate carboxykinase (PEPCK) was used for epitope prediction, resulting in one high-scoring patch centered at residue L247. NC41 synthetic peptide reached high diagnostic performance (Se 97.5% and Sp 97.5%, LR+ 39 and AUC 0.997). Data from diagnostic parameters and in silico analyses proved the usefulness of NC41 synthetic peptide as a diagnostic marker for human NC.

Albendazole Sulfoxide Plasma Levels and Efficacy of Antiparasitic Treatment in Patients With Parenchymal Neurocysticercosis.

BACKGROUND: The efficacy of albendazole therapy in patients with parenchymal neurocysticercosis (NCC) is suboptimal. Plasma levels of albendazole sulfoxide (ASOX), the active metabolite of albendazole, are highly variable among patients. We hypothesized that high ASOX plasma levels during albendazole therapy may be associated with an increased antiparasitic efficacy. METHODS: ASOX plasma levels were measured at treatment day 7 in 118 patients with parenchymal NCC enrolled in a treatment trial. The relationships between increasing ASOX plasma levels with the proportion of cysts resolved and the proportion of patients with complete cyst resolution (evaluated by 6-month brain magnetic resonance) were assessed. RESULTS: There was a trend toward a higher proportion of cysts resolved and a higher proportion of patients cured with increasing quartiles of ASOX plasma levels. In patients with 3 or more brain cysts, the regression analysis adjusted by the concomitant administration of praziquantel (PZQ) showed a 2-fold increase in the proportion of cysts resolved (risk ratio [RR], 1.98; 95% confidence interval [CI], 1.01-3.89; P = .048) and 2.5-fold increase in the proportion of patients cured (RR, 2.45; 95% CI, .94-6.36; P = .067) when ASOX levels in the highest vs the lowest quartile were compared. No association was found in patients with 1-2 brain cysts. CONCLUSIONS: We suggest an association between high ASOX plasma levels and increased antiparasitic efficacy in patients with parenchymal NCC. Nonetheless, this association is also influenced by other factors including parasite burden and concomitant administration of PZQ. These findings may serve to individualize and/or adjust therapy schemes to avoid treatment failure.

The status and health burden of neurocysticercosis in Mbulu district, northern Tanzania.

OBJECTIVE: The objective of this study was to assess the extent and health burden of neurocysticercosis in the general community of the Mbulu district, northern Tanzania. About 1051 randomly select participants were screened for human cysticercosis. The Cysticercus Western Blot IgG and Computed Tomography scan were used to detect infection by cysticerci. The DALYs was used to assess the community's health burden vis-a-vis neurocysticercosis. RESULTS: The sero-prevalence of HCC was 16.27%. About 76% of 25 selected human cysticercosis sero-positives had neurocysticercosis suggestive lesions on CT scan and 74% had history of epilepsy. Epilepsy caused 2.8 years of life lost and 2.2 healthy years of life lost due to disability per 1000 person-years in Mbulu. The average DALYs imposed due to neurocysticercosis and epilepsy were 3.0 and 3.9 per 1000 person-years, respectively. Neurocysticercosis is a serious public health concern in northern Tanzania.

[Study on diagnosis of atypical cerebral cysticercosis].

OBJECTIVE: To analyze the characteristics of the results of serum immunological tests and brain CT image examinations of atypical cerebral cysticercosis patients, so as to provide the reference for improving the diagnosis of the disease. METHODS: Totally 446 suspected cerebral cysticercosis patients were chosen as the study objects, all of them were given experimental treatment with praziquantel, and then the patients with atypical cerebral cysticercosis were diagnosed according to the treatment effect and review results of brain CT or MRI. Meanwhile, all the 446 patients were tested for serum specific IgG and IgG4 antibodies and cysticercus circulating antigen (CAg) by ELISA, McAb ELISA and PEG-ELISA respectively, and the IHA test was also performed. All the patients received the brain CT examinations. The test results were analyzed statistically and the test performances of the methods above-mentioned were calculated. RESULTS: Among the 446 suspected cerebral cysticercosis patients, after the praziquantel treatment, there were 315 patients whose symptoms were alleviated, and they were diagnosed as atypical cerebral cysticercosis. Among the 446 suspected cerebral cysticercosis patients, the positive rates of specific IgG and IgG4 antibodies, and CAg were 15.47%, 15.02%, and 11.21% respectively, and the positive rate of IHA was 28.47% (χ2 = 52.45, P ＜ 0.01). The brain CT examinations showed that there were 79.14% (353/446) of patients with suspected cysticercus foci. The sensitivities of ELISA, McAb-ELISA, PEG-ELISA, IHA test and brain CT examination (suspected cysticercus foci) for the diagnosis of atypical cerebral cysticercosis were 15.36%-96.82%, the specificities were 63.36%-99.24%, the positive predictive values were 86.40%-98.52%, the negative predictive values were 32.83%-98.25%, the positive likelihood ratios were 2.64-27.86, the negative likelihood ratios were 0.05-0.81, and the OR values were 7.16-52.80. The consolidation of the five tests above-mentioned showed the OR value was 108.00, which was 2 times of the OR value of CT examination. CONCLUSIONS: Atypical cerebral cysticercosis is definitely diagnosed mainly by means of evaluating the effect of the diagnostic therapy (anti-cysticercus), and the serum immunological examinations and brain CT examination also have important reference values.

Distinguishing neurocysticercosis epilepsy from epilepsy of unknown etiology using a minimal serum mass profiling platform.

Neurocysticercosis is associated with epilepsy in pig-raising communities with poor sanitation. Current internationally recognized diagnostic guidelines for neurocysticercosis rely on brain imaging, a technology that is frequently not available or not accessible in areas endemic for neurocysticercosis. Minimally invasive and low-cost aids for diagnosing neurocysticercosis epilepsy could improve treatment of neurocysticercosis. The goal of this study was to test the extent to which patients with neurocysticercosis epilepsy, epilepsy of unknown etiology, idiopathic headaches and among different types of neurocysticercosis lesions could be distinguished from each other based on serum mass profiling. For this, we collected sera from patients with neurocysticercosis-associated epilepsy, epilepsy of unknown etiology, recovered neurocysticercosis, and idiopathic headaches then performed binary group comparisons among them using electrospray ionization mass spectrometry. A leave one [serum sample] out cross validation procedure was employed to analyze spectral data. Sera from neurocysticercosis patients was distinguished from epilepsy of unknown etiology patients with a p-value of 10-28. This distinction was lost when samples were randomized to either group (p-value = 0.22). Similarly, binary comparisons of patients with neurocysticercosis who has different types of lesions showed that different forms of this disease were also distinguishable from one another. These results suggest neurocysticercosis epilepsy can be distinguished from epilepsy of unknown etiology based on biomolecular differences in sera detected by mass profiling.

Treatment-Resistant Human Extraparenchymal Neurocysticercosis: An Immune-Inflammatory Approach to Cysticidal Treatment Outcome.

OBJECTIVE: The aim of this study is to analyze the immune-endocrine profile in neurocysticercosis (NC) patients resistant to cysticidal treatment. METHODS: The inflammatory and regulatory responses of 8 resistant NC patients with extraparenchymal parasites and 5 healthy controls were evaluated through flow cytometry. Serum interleukin levels were measured by ELISA and catecholamines levels by high performance liquid chromatography. RESULTS: Higher percentages of Tr1, CD4+CD25+FOXP3+CD127- and CD4+CD45RO+FOXP3HI were found in NC patients compared with healthy controls, but no difference was found in catecholamine levels. Antigen-specific proliferative immune response was observed in NC patients. Neither anti-inflammatory nor pro-inflammatory cytokines showed differences between patients and controls, but IL-6 levels were lower in treatment-resistant NC patients. In addition, TGFß showed a significant negative correlation with dopamine. CONCLUSIONS: Altogether, these results may point to a modulation of the neuroinflammation in these patients that could indirectly favor cysticercal survival in CNS microenvironment.

Low sensitivity and frequent cross-reactions in commercially available antibody detection ELISA assays for Taenia solium cysticercosis.

OBJECTIVE: To evaluate the diagnostic performance of two commercially available ELISA kits, Novalisa® and Ridascreen® , for the detection of antibodies to Taenia solium, compared to serological diagnosis of neurocysticercosis (NCC) by LLGP-EITB (electro-immunotransfer blot assay using lentil-lectin purified glycoprotein antigens). METHODS: Archive serum samples from patients with viable NCC (n = 45) or resolved, calcified NCC (n = 45), as well as sera from patients with other cestode parasites (hymenolepiasis, n = 45 and cystic hydatid disease, n = 45), were evaluated for cysticercosis antibody detection using two ELISA kits, Novalisa® and Ridascreen® . All NCC samples had previously tested positive, and all samples from heterologous infections were negative on LLGP-EITB for cysticercosis. Positive rates were calculated by kit and sample group and compared between the two kits. RESULTS: Compared to LLGP-EITB, the sensitivity of both ELISA assays to detect specific antibodies in patients with viable NCC was low (44.4% and 22.2%), and for calcified NCC, it was only 6.7% and 4.5%. Sera from patients with cystic hydatid disease were highly cross-reactive in both ELISA assays (38/45, 84.4%; and 25/45, 55.6%). Sera from patients with hymenolepiasis cross-reacted in five cases in one of the assays (11.1%) and in only one sample with the second assay (2.2%). CONCLUSIONS: The performance of Novalisa® and Ridascreen® was poor. Antibody ELISA detection cannot be recommended for the diagnosis of neurocysticercosis.

Soluble Fas/FasLare elevated in the serum and cerebrospinal fluid of patients with neurocysticercosis.

Neurocysticercosis (NCC) caused by Taeniasolium is one of the most common parasitic diseases of the central nervous system. Inflammation and apoptosis are two main responses involved in NCC pathogenesis. We aimed to examine apoptosis by the TUNEL assay and apoptosis-associated sFas and sFasL levels in the cerebrospinal fluid (CSF) and serum of patients with NCC. Brain biopsy (n = 1), CSF (n = 14), and serum (n = 36) of patients with NCC and uninfected controls (n = 14 and 24 for CSF and serum, respectively) were collected together with clinical data. Residual brain tissue was analyzed by the TUNEL assay. sFas and sFasL in CSF samples and sFas, sFasL, and p53 in serum samples were measured by ELISA. Immunohistochemistry of the biopsy indicated the presence of vimentin-positive arachnoid tissue in the TUNEL-positive region. Compared to controls, sFas was significantly reduced in CSF samples of patients with NCC (P = 0.018), especially among those without inflammation, but significantly increased in the serum samples of the vesicular(P = 0.011), moderate(P = 0.025), and non-epilepsy(P = 0.049) subgroups of patients with NCC. sFasL was elevated in the CSF (P < 0.0001), as well as in the calcified subgroup (P = 0.031), but sFasL levels in CSF were similar among patients with NCC with and without inflammation. These findings support a role of sFas and sFasL in the induction of apoptosis in patients with NCC, with sFas probably being involved in the inflammation phase of NCC and depending on host factors such as parasite stage, disease severity, and symptoms, and sFasL being involved in the inflammation, non-inflammation, and calcification phase of the disease.

Experimental neurocysticercosis: absence of IL-4 induces lower encephalitis.

METHODS: BALB/c (WT) and BALB/c (IL-4-KO) mice were inoculated intracranially with Taenia crassiceps cysticerci and euthanized at 7, 30, 60 and 90 days later, the encephala removed and histopathologically analyzed. RESULTS: The absence of IL-4 induced greater parasitism. In the initial phase of the infection, IL-4-KO showed a lower intensity in the inflammatory infiltration of polimorphonuclear cells in the host-parasite interface and intra-parenquimatous edema. The IL-4-KO animals, in the late phase of the infection, showed lower intensity of ventriculomegaly, encephalitis, and meningitis, and greater survival of the parasites in comparison with the WT animals. CONCLUSION: The absence of IL-4 induced lower inflammatory infiltration, ventriculomegaly and perivasculitis in experimental NCC.

Experimental neurocysticercosis: absence of IL-4 induces lower encephalitis / Neurocisticercose experimental: ausência de IL-4 induz menos encefalite

ABSTRACT Neurocysticercosis (NCC) is the most severe clinical manifestation of cysticercosis. One of the factors responsible for its symptomatology is the host inflammatory response. Therefore the influence of interleukin 4 (IL-4) on the induction of encephalitis in experimental NCC was evaluated. Methods BALB/c (WT) and BALB/c (IL-4-KO) mice were inoculated intracranially with Taenia crassiceps cysticerci and euthanized at 7, 30, 60 and 90 days later, the encephala removed and histopathologically analyzed. Results The absence of IL-4 induced greater parasitism. In the initial phase of the infection, IL-4-KO showed a lower intensity in the inflammatory infiltration of polimorphonuclear cells in the host-parasite interface and intra-parenquimatous edema. The IL-4-KO animals, in the late phase of the infection, showed lower intensity of ventriculomegaly, encephalitis, and meningitis, and greater survival of the parasites in comparison with the WT animals. Conclusion The absence of IL-4 induced lower inflammatory infiltration, ventriculomegaly and perivasculitis in experimental NCC.

RESUMO A Neurocisticercose (NCC) é a manifestação clínica mais severa da cisticercose, e um dos fatores responsáveis pela sintomatologia é a resposta inflamatória do hospedeiro. Desta forma avaliou-se a influência da interleucina 4 (IL-4) na indução de encefalite na NCC experimental. Métodos Camundongos das linhagens BALB/c (WT) e BALB/c (IL-4-KO) foram inoculados intracranialmente com cisticercos de Taenia crassiceps e eutanasiados aos 7, 30, 60 e 90 dias após a infecção, os encéfalos foram removidos e analisados histopatologicamente. Resultados A ausência da IL-4 induziu um maior parasitismo nos animais. Na fase inicial da infecção os animais IL-4-KO apresentaram menor intensidade tanto de infiltrado inflamatório de polimorfonucleares na interface parasito-hospedeiro quanto de edema intraparenquimatoso. Os animais IL-4-KO, na fase tardia, apresentaram menor intensidade de ventriculomegalia, encefalite, meningite e maior sobrevivência dos cisticercos em relação aos animais WT. Conclusão A ausência da IL-4 induz menos infiltrado inflamatório, ventriculomegalia e perivasculite na NCC experimental.

Human Glutathione S-Transferase Enzyme Gene Polymorphisms and Their Association With Neurocysticercosis.

Neurocysticercosis (NCC), caused by cysticerci of Taenia solium is the most common helminthic infection of the central nervous system. Some individuals harboring different stages of cysticerci in the brain remain asymptomatic, while others with similar cysticerci lesions develop symptoms and the reasons remain largely unknown. Inflammatory response to antigens of dying parasite is said to be responsible for symptomatic disease. Reactive oxygen species (ROS) that are generated in inflammatory conditions can damage cellular macromolecules such as lipids, DNA, and proteins. The glutathione S-transferases (GSTs) are critical for the protection of cells from ROS. A total of 250 individuals were included in the study: symptomatic NCC = 75, asymptomatic NCC = 75, and healthy controls = 100. The individuals carrying the deletions of GSTM1 and GSTT1 were at risk for NCC (OR = 2.99, 95 %CI = 1.31-6.82, p = 0.0073 and OR = 1.94, 95 %CI = 0.98-3.82, p = 0.0550 respectively). Further, the individuals with these deletions were more likely to develop symptomatic disease (OR = 5.08, 95 % CI = 2.12-12.18, p = 0.0001 for GSTM1 and OR = 3.25, 95 %CI = 1.55-6.82, p = 0.0018 for GSTT1). Genetic variants of GSTM3 and GSTP1 were not associated with NCC. The total GST activity and levels of GSTM1, GSTT1, and GSTM3 were significantly higher in asymptomatic subjects than in symptomatic and healthy controls. Lower GST activity was observed in individuals with GSTM1 and GSTT1 deletions. The present study suggests that the individuals with GSTM1 and GSTT1 deletions are at higher risk to develop symptomatic disease. The higher GST activity and levels of GSTM1, GSTT1, and GSTM3 are likely to play role in maintaining asymptomatic condition.

Usefulness of gel filtration fraction as potential biomarker for neurocysticercosis in serum: towards a new diagnostic tool.

There is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting. Sensitivity (Se), specificity (Sp) and other diagnostic parameters were calculated. Polypeptides of interest in the diagnosis of human NCC present at GFF were analysed by mass spectrometry (MS) and B-cell epitopes were predicted. GFF had the best diagnostic parameters: Se 93·3%; Sp 93%; AUC 0·990; LR+ = 13·42 and LR- = 0·07, and proved to be useful reacting with serum samples in immunoblotting. Proteic profile ranged from 64 to 68 kDa and enolase and calcium binding protein calreticulin precursor were identified after MS. The enolase and calcium-binding protein calreticulin precursor showed 18 and 10 predicted B-cell epitopes, respectively. In conclusion we identified important markers in the GFF with high efficiency to diagnose NCC.

High Prevalence of Asymptomatic Neurocysticercosis in an Endemic Rural Community in Peru.

BACKGROUND: Neurocysticercosis is a common helminthic infection of the central nervous system and an important cause of adult-onset epilepsy in endemic countries. However, few studies have examined associations between neurologic symptoms, serology and radiographic findings on a community-level. METHODOLOGY: We conducted a population-based study of resident's ≥2 years old in a highly endemic village in Peru (pop. 454). We applied a 14 -question neurologic screening tool and evaluated serum for antibodies against Taenia solium cysticercosis using enzyme-linked immunoelectrotransfer blot (LLGP-EITB). We invited all residents ≥18 years old to have non-contrast computerized tomography (CT) of the head. PRINCIPAL FINDINGS: Of the 385 residents who provided serum samples, 142 (36.9%) were seropositive. Of the 256 residents who underwent CT scan, 48 (18.8%) had brain calcifications consistent with NCC; 8/48 (17.0%) reported a history of headache and/or seizures. Exposure to T. solium is very common in this endemic community where 1 out of 5 residents had brain calcifications. However, the vast majority of people with calcifications were asymptomatic. CONCLUSION: This study reports a high prevalence of NCC infection in an endemic community in Peru and confirms that a large proportion of apparently asymptomatic residents have brain calcifications that could provoke seizures in the future.

Intranasal microemulsion for targeted nose to brain delivery in neurocysticercosis: Role of docosahexaenoic acid.

Intranasal Microemulsions (MEs) for nose to brain delivery of a novel combination of Albendazole sulfoxide (ABZ-SO) and Curcumin (CUR) for Neurocysticercosis (NCC), a brain infection are reported. MEs prepared by simple solution exhibited a globule size <20nm, negative zeta potential and good stability. The docosahexaenoic acid (DHA) ME revealed high and rapid ex vivo permeation of drugs through sheep nasal mucosa. Intranasal DHA ME resulted in high brain concentrations and 10.76 (ABZ-SO) and 3.24 (CUR) fold enhancement in brain area-under-the-curve (AUC) compared to intravenous DHA MEs at the same dose. Direct nose to brain transport (DTP) of >95% was seen for both drugs. High drug targeting efficiency (DTE) to the brain compared to Capmul ME and drug solution (P<0.05) suggested the role of DHA in aiding nose to brain delivery. Histopathology study confirmed no significant changes. High efficacy of ABZ-SO: CUR (100:10ng/mL) DHA ME in vitro on Taenia solium cysts was confirmed by complete ALP inhibition and disintegration of cysts at 96h. Considering that the brain concentration at 24h was 1400±160.1ng/g (ABZ-SO) and 120±35.2ng/g (CUR), the in vitro efficacy seen at a 10 fold lower concentration of the drugs strongly supports the assumption of clinical efficacy. The intranasal DHA ME is a promising delivery system for targeted nose to brain delivery.

Pediatric neurocysticercosis: usefulness of antibody response in cysticidal treatment follow-up.

Serum and urine samples were collected from 33 NCC patients before the albendazole treatment, 3-6 and 12 months PT. At 3 months PT, 24 (72.7%) patients had no detectable CT/MRI lesions and 9 (27.2%) patients had persistent lesions. Antibody response to crude soluble extract (CSE), excretory secretory (ES), and lower molecular mass (LMM) (10-30 KDa) antigenic fraction of T. solium cysticerci was detected in serum and urine samples by ELISA. Before the treatment, out of 33 NCC children, 14 (42.4%), 22 (66.6%), and 11 (33.3%) serum samples were found positive with the use of CSE, ES, and LMM antigen, respectively. At 3-6 months PT, positivity rate was 5 (15.1%), 2 (6%), and 4 (12.1%) and at 12 months PT, positivity rate was 5 (15.1%), 0, and 3 (9%) with the use of CSE, ES, and LMM antigen, respectively. There was no significant difference in the positivity with the use of three antigens in pretreatment and PT urine samples. The study suggests that the use of ES antigen to detect antibody in serum samples may serve better purpose to evaluate the therapeutic response in patients with NCC.

Diethylaminoethyl (DEAE) binding fraction from Taenia solium metacestode improves the neurocysticercosis serodiagnosis.

Neurocysticercosis (NC) is one of the most important diseases caused by parasites affecting the central nervous system. We fractionated by ion-exchange chromatography using diethylaminoethyl (DEAE)-sepharose resin the total saline extract (S) from Taenia solium metacestodes and evaluated obtained fractions (DEAE S1 and DEAE S2) by enzyme-linked immunosorbent assay (ELISA, n = 123) and immunoblotting (IB, n = 22) to detect human NC in serum. Diagnostic parameters were established by ROC and TG ROC curves for ELISA tests. IB was qualitatively analyzed. S and DEAE S1 presented sensitivity of 87. 5% and DEAE S2 90%. The best specificity was observed for DEAE S2 (90.4%). In IB, using DEAE S2 samples from NC patients presented bands of 20-25, 43-45, 55-50, 60-66, 82, 89, and 140 kDa. The great diagnostic parameters reached by DEAE S2 suggest the potential applicability of this fraction in NC immunodiagnosis.

Recombinant protein- and synthetic peptide-based immunoblot test for diagnosis of neurocysticercosis.

One of the most well-characterized tests for diagnosing neurocysticercosis (NCC) is the enzyme-linked immunoelectrotransfer blot (EITB) assay developed at the CDC, which uses lentil lectin-bound glycoproteins (LLGP) extracted from Taenia solium cysticerci. Although the test is very reliable, the purification process for the LLGP antigens has been difficult to transfer to other laboratories because of the need for expensive equipment and technical expertise. To develop a simpler assay, we previously purified and cloned the diagnostic glycoproteins in the LLGP fraction. In this study, we evaluated three representative recombinant or synthetic antigens from the LLGP fraction, individually and in different combinations, using an immunoblot assay (recombinant EITB). Using a panel of 249 confirmed NCC-positive and 401 negative blood serum samples, the sensitivity of the recombinant EITB assay was determined to be 99% and the specificity was 99% for diagnosing NCC. We also tested a panel of 239 confirmed NCC-positive serum samples in Lima, Peru, and found similar results. Overall, our data show that the performance characteristics of the recombinant EITB assay are comparable to those of the LLGP-EITB assay. This new recombinant- and synthetic antigen-based assay is sustainable and can be easily transferred to other laboratories in the United States and throughout the world.

Three cases of imported neurocysticercosis in Northern Italy.

BACKGROUND: Neurocysticercosis (NCC) is an important cause of adult-onset seizures in endemic areas, whereas it is emerging in some nonendemic areas as well because of extensive immigration. METHOD: We describe three cases of imported NCC recently admitted to San Bortolo Hospital in Vicenza, located in Northern Italy. RESULTS: All patients were immigrants. One patient was human immunodeficiency virus positive with severe immunosuppression. The diagnosis of NCC was made on the basis of magnetic resonance results; failure of anti-Toxoplasma, antitubercular, and antifungal therapy; and regression of the cystic lesions after empiric therapy with albendazole. Serology was positive in only one case. In one patient, NCC was diagnosed by biopsy of the brain lesion. CONCLUSION: In nonendemic countries, NCC should be included in the differential diagnosis of all patients coming from endemic areas with seizures, hydrocephalus, and compatible lesions on brain imaging. Long-term follow-up is required but may be difficult to implement because these patients tend to move in search of employment. Screening of patient's household contacts for Taenia solium infection should always be carried out.