Immunoblot Kit Research Articles

Capsaicin induces ATP-dependent thermogenesis via the activation of TRPV1/β3-AR/α1-AR in 3T3-L1 adipocytes and mouse model

Capsaicin (CAP) is a natural bioactive compound in chili pepper that activates the transient receptor potential vanilloid subfamily 1 (TRPV1) and is known to stimulate uncoupling protein 1 (UCP1)-dependent thermogenesis. However, its effect on ATP-dependent thermogenesis remains unknown. In this study, we employed qRT-PCR, immunoblot, staining method, and assay kit to investigate the role of CAP on ATP-dependent thermogenesis and its modulatory roles on the TRPV1, β3-adrenergic receptor (β3-AR), and α1-AR using in vitro and in vivo models. The studies showed that CAP treatment in high-fat diet-induced obese mice resulted in lower body weight gain and elevated ATP-dependent thermogenic effectors' protein and gene expression through ATP-consuming calcium and creatine futile cycles. In both in vitro and in vivo experiments, CAP treatment elevated the protein and gene expressions of sarcoendoplasmic/endoplasmic reticulum calcium ATPase 2 (SERCA2), ryanodine receptor 2 (RYR2), creatine kinase B (CKB), and creatine kinase mitochondrial 2 (CKMT2) mediated by the activation of β3-AR, α1-AR, and TRPV1. Our study showed that CAP increased intracellular Ca2+ levels and the expression of voltage-dependent anion channel (VDAC) and mitochondrial calcium uniporter (MCU) which indicates that increased mitochondrial Ca2+ levels lead to increased expression of oxidative phosphorylation protein complexes as a result of ATP-futile cycle activation. A mechanistic study in 3T3-L1 adipocytes revealed that CAP induces UCP1- and ATP-dependent thermogenesis mediated by the β3-AR/PKA/p38MAPK/ERK as well as calcium-dependent α1-AR/TRPV1/CaMKII/AMPK/SIRT1 pathway. Taken together, we identified CAP's novel functional and modulatory roles in UCP1- and ATP-dependent thermogenesis, which is important for developing therapeutic strategies for combating obesity and metabolic diseases.

Polystyrene microplastics mediate cell cycle arrest, apoptosis, and autophagy in the G2/M phase through ROS in grass carp kidney cells.

Microplastics (MPs) have attracted widespread worldwide attention as a new pollutant. However, the role of reactive oxygen species (ROS) and cell cycle in nephrotoxicity induced by different concentrations of polystyrene microplastics (PS-MPs) is unknown. This study used grass carp kidney cells (CIK) treated with different concentrations of PS-MPs (0, 0.012, 0.0625, and 0.5 mg L-1 ) as subjects. With the increase of PS-MPs concentration, the levels of ROS and malonaldehyde increased, while the level of total antioxidant capacity, superoxide Dismutase (SOD), and glutathione (GSH) activity decreased. The expression of BUB1 mitotic checkpoint serine/threonine kinase (BUB1), cyclin-dependent kinase (CDK1), CDK2, CyclinB1, cell division cycle 20 homolog (CDC20), and B-cell lymphoma-2, sequestosome 1 decreased significantly. Nevertheless, the expression of Caspase 3, Cleave-Caspase 3, cytochrome c (Cytc), BCL2-associated X, apoptosis regulator, poly ADP-ribose polymerase (PARP), Cleave-PARP, Caspase 9, autophagy immunoblot kit (LC3), and Beclin1 increased. Our research shows that PS-MPs can trigger oxidative stress and induce cell cycle arrest, apoptosis, and autophagy in CIK cells by regulating ROS. This work provides a theoretical basis for cellular biology and toxicology mechanisms and new insights into the potential risks to animals from MPs exposure in the environment.

LARP7 upregulates SIRT1 deacetylase activity and inhibits Th1/Th17 cytokine response in psoriatic mice

Objective: To investigate the possible role of La ribonucleoprotein 7 (LARP7) in psoriasis through a mouse model and uncover its underlying mechanism. Methods: The back skin of C57BL/6 mice was smeared with IMquimod (IMQ) cream for 7 days to induce psoriasis. Immunoblot kit was used to detect the deacetylase activity of SIRT1 (member of sirtuin family). Hematoxylin and eosin staining was used to assess the degree of psoriasis in mouse. Flow cytometry assays were performed to confirm effects on Th1/Th17 cell differentiation. Enzyme-linked-immunosorbent serologic assays were used to detect the level of secreted cytokines. Results: LARP7 upregulated SIRT1 deacetylase activity. LARP7 alleviated psoriasis symptoms in mice by upregulating SIRT1 deacetylase activity. In addition, LARP7 regulated Th1/Th17 cell differentiation in psoriatic mice. We further found that LARP7 inhibited Th1/Th17 cytokine. Conclusion: LARP7 upregulated SIRT1 activity and inhibited Th1/Th17 cytokine response in psoriatic mice.

LARP7 upregulates SIRT1 deacetylase activity and inhibits Th1/Th17 cytokine response in psoriatic mice.

To investigate the possible role of La ribonucleoprotein 7 (LARP7) in psoriasis through a mouse model and uncover its underlying mechanism. The back skin of C57BL/6 mice was smeared with IMquimod (IMQ) cream for 7 days to induce psoriasis. Immunoblot kit was used to detect the deacetylase activity of SIRT1 (member of sirtuin family). Hematoxylin and eosin staining was used to assess the degree of psoriasis in mouse. Flow cytometry assays were performed to confirm effects on Th1/Th17 cell differentiation. Enzyme-linked-immunosorbent serologic assays were used to detect the level of secreted cytokines. LARP7 upregulated SIRT1 deacetylase activity. LARP7 alleviated psoriasis symptoms in mice by upregulating SIRT1 deacetylase activity. In addition, LARP7 regulated Th1/Th17 cell differentiation in psoriatic mice. We further found that LARP7 inhibited Th1/Th17 cytokine. LARP7 upregulated SIRT1 activity and inhibited Th1/Th17 cytokine response in psoriatic mice.

Significance of total and specific IgE in asthma of Iraqi adult patients

A case-control retrospective study was conducted during January – June 2019 on 104 patients with asthma (50 females and 54 males) and 111 non-asthmatic healthy subjects matched patients for gender (54 females and 57 males) to assess role of total and specific IgE serum levels in patho-physiology of disease using ELISA and multiplex immunoblot kits. The mean age ± standard deviation of asthma patients was 37.9 ± 12.1 years, while it was lower among control (35.9 ± 10.5 years), but the difference was not significant. Males and females were similarly distributed among asthma cases (48.1 and 51.9%, respectively) and control (48.6 and 51.4%, respectively). Most of asthma cases were observed to have mild disease (49%), followed by moderate and severe asthma (93.4 and 11.5%, respectively). It was also observed that 51.9% of patients had a family history of asthma. With respect to allergen type, 21.2% of asthma cases were seropositive for mixed types of allergens. Median level of total IgE was significantly (p < 0.001(elevated in sera of asthma cases (204.1 ng/ml) compared to control subjects (163.3 ng/ml). The level positively paralleled the severity of disease. The highest level was observed in severe cases (244.9ng/ml, while the lowest level was recorded in mild cases (189.7 ng/ml). The level of specific IgE showed some variations among asthma cases distributed according to the type of allergen, the highest median was recorded in seronegative cases (215.2 ng/ml), while a lowest level was observed in cases seropositive for animal dander (189.3 ng/ml). Logistic regression analysis revealed that total asthma patients, as well as male and female cases were at an increased risk to develop asthma episodes due to elevated serum level of IgE.ROC analysis demonstrated that elevated serum level of IgE occupied a significant (p = 0.001) area under curve (AUC), which was 0.828. In conclusion, the present study confirms the significant role of total and specific IgE in patho-physiology of bronchial asthma and its correlation with disease severity and allergen type in adults

Associations of autoantibodies and clinical profile of the patients with systemic sclerosis

AbstractBackgroundSystemic sclerosis is characterized by the involvement of organs and the presence of specific antibodies. The objectives of this study were to identify the autoantibodies and to determine their association with the selected clinical features of the disease among Bangladeshi systemic sclerosis patients.MethodsThis cross‐sectional study was performed at the rheumatology outpatient clinic of Bangabandhu Sheikh Mujib Medical University. Autoantibodies against nine systemic sclerosis‐specific antigens were tested using an enzyme‐linked immunoassay immunoblot kit. Several clinical features of patients with positive and negative autoantibody were examined by χ2 or Fisher's exact tests.ResultsA total of 71 patients with systemic sclerosis (66; 93.0% female) were included. Their mean age at disease onset was 33.2 years. Fifty‐seven (80.3%) patients had diffuse cutaneous subtype. Out of nine autoantibodies, four were positive, anti‐topoisomerase‐I (57.7%), anti‐U1 ribonucleic protein (21.1%), anti‐RNA polymerase III (18.3%), and anticentromere antibodies (4.2%). Eleven (15.5%) patients were negative for any antibodies and 11 patients were positive for at least two autoantibodies. Anti‐U3‐RNP, anti‐PM‐Scl, anti‐Ku, and anti‐Th/To auto antibodies were absent in all patients. Anti‐RNA polymerase III was associated with raised pulmonary arterial systolic pressure (PASP) and anti‐U1‐RNP with decreased forced vital capacity (FVC).ConclusionsAnti‐topoisomerase‐I was the commonest autoantibody in patients with systemic sclerosis in Bangladesh. Anti‐RNA polymerase III antibody had significant association with raised PASP and anti‐U1‐RNP with decreased FVC.

Scleroderma-specific autoantibodies: Should they be included in the diagnostic work-up for Sjögren's syndrome?

ObjectivesSicca complaints are a frequent reason for rheumatologic consultation. Testing for specific antibodies against Ro/SSA and La/SSB antigens and minor salivary gland (MSG) biopsy are among the main tools implemented in the diagnostic work-up. Anticentromere antibodies and sicca manifestations are frequently detected in Sjögren's syndrome (SS) and systemic sclerosis (SSc), respectively. Herein, we aimed to determine the frequency and clinical associations of a wide spectrum of scleroderma (SSc)-specific autoantibodies in consecutive patients referred for evaluation of possible SS. MethodsDemographic, clinico-pathological, and laboratory data were recorded in 216 consecutive patients with sicca complaints. All study participants were tested for SSc-specific autoantibodies (against CENP, PM/Scl, Scl-70, Ku, NOR90, RP11, RP155, fibrillarin, PDGFR, and Th/To) using a commercially available immunoblot kit. According to band intensity, the identified autoantibodies were further classified in those with strong and medium titers. ResultsSSc-specific autoantibodies were detected in 41.7% (90/216) patients evaluated (19% at strong, 22.7% at medium titers) without significant differences between anti-Ro/SSA positive and negative groups. At strong titers was significantly higher in patients with MSG biopsies fulfilling SS histopathological criteria (30% vs 12.5%, p = 0.009). This association remained significant after adjustment for antibodies against Ro/SSA and La/SSB autoantigens [OR 95% (CI): 4.1 (1.5–10.6)]. ConclusionSSc-specific autoantibodies are frequently detected among patients presenting with sicca complaints and at strong but not medium titers are independently associated with MSG biopsy positivity. Taken together, these data imply a useful role of SSc antibody testing in the diagnostic work-up and possibly in the classification criteria for SS.

Calpain-mediated cleavage of mitochondrial fusion/fission proteins in acetaminophen-induced mice liver injury

Mitochondrial dysfunction was considered to be a critical event in acetaminophen (APAP) -induced hepatotoxicity. Recent studies suggest that abnormal mitochondrial dynamics contributes to mitochondrial dysfunction in APAP-induced liver injury, yet the underlying mechanisms responsible for deregulated mitochondrial dynamics remains elusive. In this study, C57BL/6 mice were used to establish a model of acute liver injury via intraperitoneal (i.p.) injection with overdose of APAP. Furthermore, calpain intervention experiments were achieved by the inhibitors ALLN or calpeptin. The activity of serum enzymes and pathological changes of APAP-treated mice were evaluated, and the critical molecules in mitochondrial dynamics and calpain degradative pathway were determined by electron microscopy, immunoblot and calpain activity kit. The results demonstrated that APAP overdose resulted in a severe liver injury, mitochondrial damage and an obvious cleavage of fusion/fission proteins. Meanwhile, the activation of calpain degradative machinery in liver were observed following APAP. By contrast, pretreatment of calpain inhibitors significantly inhibited the activation of calpains. Our further investigation found that ALLN or calpeptin administration significantly suppresses the changes of mitochondrial dynamics in APAP-treated mice and finally protected against APAP-induced hepatoxicity. Overall, these results suggest that calpain-mediated cleavage of mitochondrial dynamics proteins was involved in the pathogenic process of mitochondrial dysfunction and thus present a potential molecular coupling APAP-induced hepatotoxicity.

The Role of IL-5, IL-33 and Total IgE in a Sample of Workers Suffering from Paints and Mills Asthmatic Patients

A case-control retrospective study was conducted during November 2020 on 88 asthmatic patients (38 male mill and 50 male paints workers)with age ranged between (15-25),and 44 control with age ranged between (15-25)years to assess the role of total IgE ,interleukine5(IL-5),and interleukin33(IL-33)serum levels in pathophysiology of disease using ELISA and multiplex immunoblot kits. Some hematological changes such as(Eosinophil and Basophil%)by Beckman coulter. The patients’ samples were obtained from Nature Dyes Company Ltd in the industrial region of Aweerij Industrial, as well as Al-Dora flour mill in Baghdad, which is part of the general company for grain processing. The tests were carried out at the Specialized Center of Allergy in Baghdad/Al-Rusafa from October 2020 to April 2021. Specialized respiratory disorders and Asthma diagnose these cases, which are clinically diagnosed according to international guide lines. The present study confirms the highly significant role of total IgE and IL5,IL-33 levels compared with control in pathophysiology of bronchial asthma and it correlation with disease severity and allergen type in adults. Results showed that there were highly significant increase in Eosinophil and basophil % in cases asthma of paints and mills patients with control group. In addition, there were a positive correlation between Eosinophil and Basophil in asthma of paints, and in asthma of mills. There was positive correlation between total IgE with IL-5 and IL-33 in asthmatic group.

MO276TOTAL AND SPECIFIC IGE IN PATIENTS WITH IGA NEPHROPATHY

Abstract Background and Aims IgA nephropathy (IgAN) is the most common form of primary glomerulonephritis characterized by impaired immunological tolerance. One of possible trigger factors for IgAN initiation and development are allergens which have been described to form IgA complexes deposited in the mesangium. Previous studies reported that serum immunoglobulin class E (IgE) levels are elevated and suggested as a prognostic indicator in glomerular diseases. However, mechanisms of IgE/IgG4-mediated allergic reactions involvement in IgAN as well as specific allergens and their clinical significance have poorly been investigated. The aim of the study was to assess the total serum IgE and IgG4 level and to determine the allergoprofile in IgAN patients. Method The peripheral blood was obtained from 21 IgAN patients (aged of 32.0 (27.0 ÷ 36.0) y.o., male/female ratio as 14/7) and 18 donors (aged of 38.0 (30.0 ÷ 46.0) y.o., male/female ratio as 10/8). IgAN diagnosis was confirmed in patients’ biopsy materials according to the Oxford classification (MEST-C score). The concentrations of total IgE and IgG4 were determined using «IgE total-ELISA-BEST» kit («Vector-Best», RF) and «Human IgG4 Platinum ELISA» kit («eBioscience», Austria)». Allergen-specific IgE to 55 inhalation (domestic, epidermal, fungal, plant) or food allergens were measured using «EUROLINE Atopy Screen IgE» immunoblot kit («Euroimmun», Germany). Statistical analysis was done in Statistica 8.0. Results The increased level of total IgE (in 47,4% of cases) but not IgG4 was established in sera of IgAN patients as compared to donors (p=0.004). Moreover, proteinuria was lower in IgAN patients with a high IgE level as compared to normal one (610 (220 ÷ 910) mg/day vs 1545 (600 ÷ 2200) mg/day, p=0.01) as well as there were no any crescent formation. The investigation of allergen-specific IgE in IgAN patients revealed a strong positive reaction to dust mite allergens of Dermatophagoides pter. (22,75 (16,72 ÷ 35,69) IU/ml – in 29% of cases) and Dermatophagoides farinae (38,24 (12,19 ÷ 65,88) IU/ml – in 43% of cases) correlated with the total IgE (R=0,57, p=0,03) and IgM (R=0,54, p=0,04). The more concentration of specific IgE to dust mite allergens was the lower the severity of segmental glomerulosclerosis (R= -0.55, p=0.04), crescent formation (R= -0.56, p=0.03), proteinuria (R= -0.65, p=0.01) and haematuria (R = -0.55, p = 0.04) were reported. A weak positive reaction was established to other domestic (house dust, honey bee venom, common wasp venom, cockroach), epidermal (cat), plant (sweet vernal grass, orchard grass, timothy grass, cultivated rye, alder, birch, hazel, oak, common ragweed, mugwort, plantain) and food (carrot, sesame, hazelnut, apple) allergens in 9–18% of IgAN patients. Conclusion The prevailing of specific IgE to dust mite allergens of Dermatophagoides pter. and Dermatophagoides farinae but not to food or plant allergens was determined in IgAN patients. The obtained data demonstrated that IgAN pathogenesis in patients with latent sensibilization to dust mite allergens is characterized by a milder disease course what allows to identify this cohort as possible secondary form of IgAN.

Evaluation of Two Serological Screening Kits for Hepatitis C Virus Infection at the Regional Blood Transfusion Center of Ouagadougou, Burkina Faso

Introduction: In Burkina Faso, screening for hepatitis C virus in blood donations is made using sensitive ELISA (Enzyme Linked Immuno Sorbent Assay) type kits. However, no confirmation of the positive results obtained with these kits is made before their notification to the blood donors due to the high costs of the confirmation kits of immunoblots type. Objective: Evaluate two rapid kits against one immunoblot kit in order to determine the most efficiency which will be proposed as an alternative for the confirmation of ELISA tests in the socio-economic context of Burkina Faso. Material and Methods: The study was carried out using a panel of 72 sera, of which 22 were positive for anti-HCV antibodies and 50 were negative. The sera were tested using the Monolisa® HCV Ag-Ab ULTRA kit and confirmed with the DECISCAN HCV Plus kit. The panel was then tested with the SD BIOLINE HCV kit and the HCV TRI-DOT kit and the results obtained were evaluated against those of the DECISCAN HCV Plus kit used as “gold standard”. Results: Compared to the DECISCAN HCV Plus kit, the HCV TRI-DOT kit exhibited a sensitivity and specificity of 100% and the SD BIOLINE HCV kit a sensitivity of 86.36% and a specificity of 100%. Conclusion: Based on the results recorded by the HCV TRI-DOT kit, it would be best suited to the sero-epidemiological context of blood donors from the National Blood Transfusion Center and could be proposed as an alternative for confirmation of ELISA tests.

Pitfalls in the detection of CV2 (CRMP5) antibodies

CV2 antibodies (CV2-ab) associate with paraneoplastic neurological syndromes (PNS) and small-cell lung cancer. This study was designed to assess the sensitivity of two widely used anti-CV2 commercial kits. Fifty three sera with CV2-ab identified by immunohistochemistry on paraformaldehyde-perfused rat brain were tested with two commercial immunoblot kits (Euroimmun AG, and Ravo Diagnostika) and 4 (7.5%) of them were negative with the commercial kits. The 4 samples were positive by immunofluorescence on HEK293 cells transfected with CRMP5 and immunoblot of these cells lysate. A few CV2-ab-positive sera may be missed by commercial immunoblots. Negative samples from patients with high suspicion for PNS should be tested by alternative methods.

A new IgG immunoblot kit for diagnosis of toxoplasmosis in pregnant women.

The determination of the accurate immune status of pregnant women is crucial in order to prevent congenital toxoplasmosis. Equivocal results with conventional serological techniques are not uncommon when IgG titers are close to the cut-off value of the test, so that a confirmatory technique is needed. For this purpose, we developed a homemade immunoblot (IB) using soluble extract of Toxoplasma gondii tachyzoites and assessed it by testing 154 positive, 100 negative, and 123 equivocal sera obtained from pregnant women. In order to select the more valuable bands in terms of sensitivity and specificity, we used the Youden Index (YI). The highest YIs were those given by the 32, 36, 98, 21, and 33 bands. The simultaneous presence on the same blot of at least 3 bands showed a much higher YI (0.964) and was adapted as the positivity criterion. The analysis of results showed that our homemade IB correlated well with the commercial LDBIO Toxo II IgG® kit recently recommended as a confirmatory test (96.7% of concordance).

Lyme borreliosis — Lessons learnt from 25 years

We recognized the first Hungarian Lyme patients just 25 years ago, in 1984. It was exactly 20 years ago, when we opened the Lyme Disease Outpatient Service at the Central (“László”) Hospital for Infectious Diseases. Fifteen years ago, we established the financially independent Center for Tick-borne Diseases. The milestones of this work at the Center for Tick-borne Diseases are the description of a new tick-borne rickettsial illness (tick-borne lymphadenopathy), development of a Lyme immunoblot kit and an automated immunoblot reader (AIR). We described a simple and reliable method for detection of intrathecal borrelia antibody synthesis which is necessary for the diagnosis of neuroborreliosis. We also developed and routinely apply the comparative immunoblot assay for the evaluation of serological progression and/or regression, which can help the clinicians in the judgment whether a serological reaction is resulted from a previous, healed or an active borrelia infection. We studied the pregnancy outcome of borrelia infected mothers and provided the supporting data of that untreated borrelia infection results in higher chance of adverse pregnancy outcome.

Comparison of the performance of serological kits for Helicobacter pylori infection with European and Asian study populations

Most commercial kits for the detection of Helicobacter pylori were developed and validated with Western populations, and some have been found to perform less well with Asian populations. This study compared the performances of three serological kits with Swedish and Vietnamese peptic ulcer patients and asymptomatic individuals. The Pyloriset EIA-GIII and HM-CAP ELISA kits indicated that Asian populations had lower antibody titres to H. pylori than European populations. Despite the difference, the Pyloriset EIA-GIII kit performed well with Vietnamese peptic ulcer patients and population controls. The HM-CAP ELISA kit had a significantly lower performance with Asian populations that could not be improved by adjustments to the cut-off level. The Helicoblot 2.1 immunoblot kit performed equally well with Vietnamese and Swedish populations, although the response rate to the 35-kDa band was significantly lower with Vietnamese individuals.

Rapid detection of fungal endophytes in grasses for large‐scale studies

Summary Standard visual screening methods for determining the qualitative and quantitative presence of fungal endophytes are too time‐consuming for large‐scale ecological studies. We investigated whether commercially available immunoblot kits, using monoclonal antibody techniques and designed for rapid‐screening of the presence of Neotyphodium endophytes in fresh samples of the pasture grasses Festuca arundinacea and Lolium perenne, could be used for Neotyphodium detection using other grasses and preserved samples. We also determined whether immunoblot kits could provide quantitative information about the amount of Neotyphodium in the grass. The kits accurately detected endophyte presence in F. rubra, F. ovina, F. pratensis and F. altaica, in both preserved samples (dried and fixed), including 12‐year‐old stored, dried samples of F. rubra. Endophytes were detected in 7‐day‐old seedlings of Lolium perenne, 3 days (30%) earlier than previously recognized. The intensity of the coloured tissue prints on scanned immunoblot cards was significantly positively correlated with hyphal density, demonstrating a previously unrecognized accurate quantitative application. These findings greatly reduce logistical barriers to large‐scale field research into the broader ecological significance of Neotyphodium in temperate and arctic grasses in non‐agricultural ecosystems (particularly in remote areas) and suggest potential for estimating historical infection rates using stored and herbarium specimens.

Utility of a commercial immunoblot kit (BAG-Borrelia blot) in the diagnosis of the preliminary stages of Lyme disease

The aim of this study was to evaluate the usefulness of a commercial immunoblot (IgG and IgM BAG-Borrelia blot) in the serologic diagnosis of the early stages of Lyme disease. A total of 42 sera from patients with Lyme disease (24 patients with localized early stage (LES) and 18 patients with disseminated early stage (DES)) and 129 sera from patients with non-Lyme diseases (specificity control sera) were studied. IgG anti-p41 from Borrelia burgdorferi s.l. was present in 95.2% of patients followed by anti-p41/I PBi (16.7%), anti-p100 (9.5%) and anti-OspA (9.5%). IgM anti-p41 was present in 66.7% of patients, p41/iPBi (54.8%) and OspC (33.3%). IgM against p100, OspA and OspC were more frequent in DES patients (16.7%, 27.8% and 44.4%) than in LES patients (0.0%, 4.2% and 25.0%). In 4.8% of the cases no IgG bands were present and in 26.2% no IgM bands were present. With the exception of isolated p41 bands (59.5%), no band pattern exceeded 17%. Using manufacturer’s instructions, test sensitivity in diagnosis of the early stage of Lyme disease is 61.9%, specificity 98.4% and positive and negative predictive values 92.8% and 88.8% respectively. Applying the EUCALB 5, 6 or 7 rules sensitivity increased to 73.8% although specificity decreased to 89.9%. Of the 129 specific control sera, 41.8% presented IgG anti-p41 and 10.8% IgM anti-p41. Patients with non-Lyme diseases that presented more IgG and IgM bands were those patients with syphilis (88.2%), patients with anti-HIV antibodies (57.8%) and patients with anti-nuclear antibodies (ANA) (52.3%).

Evaluation of three tests for the detection of rabbit haemorrhagic disease virus in wild rabbits

Two double antibody sandwich ELISA kits and the immunoblot were evaluated for the detection of rabbit haemorrhagic disease (RHD) virus in wild rabbits. Either liver alone or liver and spleen...