Serum Enzyme-linked Immunosorbent Assay Research Articles

HIV testing among young adults and older adolescents in the setting of acute substance abuse treatment.

To examine the feasibility and acceptance of HIV testing among youth in acute substance abuse treatment. Youth, aged 18 to 25 years old, in state-funded inpatient detoxification completed a confidential demographic/risk behavior questionnaire, and were offered a choice of no testing, serum-based testing, or oral fluid (Orasure, Epitope, Inc., Beaverton, OR, U.S.A.) HIV testing. In all, 74% of 204 participants accepted HIV testing. In a multivariate model, female gender (odds ratio [OR], 0.32; 95% confidence interval [CI] 0.14-0.74) and having been recently tested (OR, 0.11; 95% CI, 0.05-0.26) were independently associated with refusing testing. Recent sexual activity (OR, 5.4; 95% CI, 1.5-20.4), recent use of methamphetamines (speed) or a combination of cocaine and heroin (speedball) (OR, 3.8; 95% CI, 1.6-9.0), and a recent perceived risk for HIV (OR, 4.6; 95% CI, 1.9-10.9) were independently associated with test acceptance. Thus, 150 of 150 (100%) chose the Orasure test. Overall, 64.6% (97 of 150) of those tested received their results, but among participants requiring a follow-up appointment to learn test results, only 9.2% (8 of 87) returned. HIV testing is feasible and acceptable in this population. All patients preferred Orasure testing to a serum enzyme-linked immunosorbent assay (ELISA). Most youth tested in detoxification will only learn their results if they are provided during treatment. Rapid HIV testing with same-day results could improve follow-up rates.

Prospective Evaluation of the Effect of Initiating Indinavir-Based Therapy on Insulin Sensitivity and B-Cell Function in HIV-Infected Patients

Objectives: To examine the feasibility and acceptance of HIV testing among youth in acute substance abuse treatment. Methods: Youth, aged 18 to 25 years old, in state-funded inpatient detoxification completed a confidential demographic/risk behavior questionnaire, and were offered a choice of no testing, serum-based testing, or oral fluid (Orasure, Epitope, Inc., Beaverton, OR, U.S.A.) HIV testing. Results: In all, 74% of 204 participants accepted HIV testing. In a multivariate model, female gender (odds ratio [OR], 0.32; 95% confidence interval [CI] 0.14-0.74) and having been recently tested (OR, 0.11; 95% CI, 0.05-0.26) were independently associated with refusing testing. Recent sexual activity (OR, 5.4; 95% CI, 1.5-20.4), recent use of methamphetamines (speed) or a combination of cocaine and heroin (speedball) (OR, 3.8; 95% CI, 1.6-9.0), and a recent perceived risk for HIV (OR, 4.6; 95% CI, 1.9-10.9) were independently associated with test acceptance. Thus, 150 of 150 (100%) chose the Orasure test. Overall, 64.6% (97 of 150) of those tested received their results, but among participants requiring a follow-up appointment to learn test results, only 9.2% (8 of 87) returned. Conclusions: HIV testing is feasible and acceptable in this population. All patients preferred Orasure testing to a serum enzyme-linked immunosorbent assay (ELISA). Most youth tested in detoxification will only learn their results if they are provided during treatment. Rapid HIV testing with same-day results could improve follow-up rates.

Epidemiology of herpes simplex virus in children by detection of specific antibodies in saliva.

To facilitate the study of the prevalence of herpes simplex virus (HSV) infection and its determinants in children, we developed a noninvasive saliva test. A capture enzyme-linked immunosorbent assay (ELISA) for the detection of IgG to HSV in saliva was developed, validated against a commercial serum ELISA in 110 children and 187 adults and used in a cross-sectional population-based study including 2,048 children ages 1 to 17 years, recruited in day-care centers and schools of Geneva, Switzerland. Demographic and socioeconomic determinants of HSV prevalence were studied. The sensitivity and specificity of the saliva assay were 94.1 and 95.5%, respectively, compared with the commercial serum ELISA. Participation in the cross-sectional study was 86.6%. The overall prevalence of anti-HSV IgG was 23.91%. It increased with age up to 7 years, reaching a plateau at 35% without evidence for day-care or school transmission. The main determinants of prevalence were region of national origin and parents' professional category. This new saliva-based assay proved its feasibility in the first population-based study of HSV prevalence in children that uses saliva, confirmed its validity by identifying determinants of prevalence consistent with previous reports and yielded new information, such as the lack of influence of day-care attendance, in the population studied.

Serum autoantibodies to heat shock proteins in glaucoma patients from Japan and the United States

Objective To study serum titers of antibodies against heat shock proteins in glaucoma patients from two different ethnic populations and to examine the relationship between serum antibody titers and glaucomatous damage. Study design Comparative, cross-sectional study. Participants Twenty-seven age-matched patients with primary open-angle glaucoma, 28 patients with normal pressure glaucoma, and a control group of 26 healthy subjects from Japan; and 21 age-matched patients with primary open-angle glaucoma, 40 patients with normal pressure glaucoma, and a control group of 20 healthy subjects from the United States. Main outcome measures Measurement of serum antibody titers and examination of optic disc and visual field parameters. Methods Serum immunoreactivity against heat shock proteins, including hsp27, αB-crystallin, and human and bacterial hsp60, was studied by enzyme-linked immunosorbent assay (ELISA). The relationship of serum antibody titers to glaucoma parameters, including mean deviation, neural rim area-to-disc area ratio, and peripapillary atrophy area-to-disc area ratio was examined. Results The serum ELISA titers of antibodies, including hsp27, αB-crystallin, human hsp60, and bacterial hsp60 antibodies, were higher in glaucoma patients compared with control subjects in both the Japanese and American groups ( P < 0.05). There was no association between the serum antibody titers and optic disc parameters in either group from Japan or the United States ( P > 0.05). Conclusions These findings suggest that increased titers of circulating antibodies against heat shock proteins occur in both Japanese and American patients with glaucoma. The lack of a relationship between the level of serum antibody titers and the degree of glaucomatous damage in either ethnic group suggests that increased antibodies to heat shock proteins do not occur as an epiphenomenon of the glaucomatous neurodegeneration process.

Helicobacter mustelae infection in ferrets: Pathogenesis, epizootiology, diagnosis, and treatment

Gastric infection by Helicobacter mustelae is extremely prevalent in domestic ferrets. Colonization of the ferret stomach by this organism results in gastritis and peptic ulcer disease and has been associated with gastric adenocarcinoma and mucosa-associated lymphoid tissue lymphoma. Histologically, H. mustelae infection of the ferret produces superficial mononuclear gastritis of the body, and diffuse antral gastritis accompanied by focal glandular atrophy of the antrum. Diagnosis of H. mustelae infection may be accomplished by culture of endoscopic biopsy samples, visualization of organisms in Warthin-Starry-stained sections of gastric tissue, evaluation of serum enzyme-linked immunosorbent assay titers to H. mustelae , fecal culture, or urea breath test. Treatment modalities typically include one or two select antibiotics along with mucosal protectants, proton pump inhibitors, and/or H-2 receptor antagonists in various combinations. Eradication is readily achieved with antimicrobial therapy of adequate duration. Studies to determine the efficacy of various treatment modalities, epizootiology, pathogenesis, and the influence of H. mustelae on gastric neoplasia have been largely stimulated by the use of the ferret as an animal model of H. pylori infection in humans.

Dose‐Dependent Effect of Ethanol on Hepatic Oxidative Stress and Interleukin‐6 Production After Burn Injury in the Mouse

Burned patients with detectable blood alcohol levels (BAL) show an elevated mortality rate. Interleukin (IL)-6 and reactive oxygen species (ROS) production is stimulated independently by alcohol and burn injury. The aim of the study was to determine whether increasing levels of alcohol differentially enhance the hepatic production of IL-6 and ROS after burn in a murine model of dorsal scald injury. Groups of mice received either saline or alcohol intraperitoneally to reach a BAL of 100 mg/dl or 300 mg/dl at the time of burn (15% total body surface scald) or sham injury. Burn injury alone resulted in a low mortality rate at 24 hr after injury as did the burn group with a BAL of 100 mg/dl (15%), whereas 57% of the mice burned with a BAL of 300 mg/dl did not survive (p = 0.02). Twenty-four hours after burn or sham injury, IL-6 levels were measured by enzyme-linked immunosorbent assay in serum and liver. In the saline-treated group, IL-6 circulating and hepatic levels rose after burn injury (p < 0.03). Circulating IL-6 levels in sham mice increased 1.5-fold in the group with a BAL of 100 mg/dl and 3-fold in those with a BAL of 300 mg/ml (p = 0.005 versus burn-injured, saline-treated). IL-6 hepatic production after burn injury was higher in the mice with a BAL of 300 mg/dl than in those with a BAL of 100 mg/dl and the saline-treated group (p = 0.001). Among the burned mice, alcohol exposure increased hepatic ROS production, measured by lipid peroxidation and protein oxidation, in a dose-dependent manner. Alcohol enhances in a dose-dependent manner the hepatic production of IL-6 induced by burn injury through the modulation of oxidative stress. The increased mortality rate of mice exposed to alcohol and burn injury may be due to the adverse effect on immune function induced by IL-6 elevation.

Western blotting is useful in the salivary diagnosis of Helicobacter pylori infection

Background—The salivary diagnosis of Helicobacter pylori infection offers attractive possibilities for the epidemiological study of infection in children. Salivary enzyme linked immunosorbent assay (ELISA) is less reliable then serum ELISA,...

Autoantibodies directed against bactericidal/permeability-increasing protein in patients with cystic fibrosis: association with microbial respiratory tract colonization.

Cystic fibrosis (CF) is associated with the appearance of serum autoantibodies directed against bactericidal/permeability-increasing protein (BPI). To determine the age-specific seroprevalence rates of anti-BPI-IgG and IgA in a population of patients with CF and to correlate anti-BPI antibody concentrations with microbial respiratory tract colonization and pulmonary function variables at the time of serum sampling and 6 years thereafter. Determination of BPI antibodies of the IgG and IgA isotypes using a commercial enzyme-linked immunosorbent assay in sera of a CF serum bank of 1992; correlation of anti-BPI antibody concentrations with age, clinical score, pulmonary function variables in 1992 and 1998, total serum immunoglobulin isotype concentrations and respiratory tract colonization with Pseudomonas aeruginosa and Aspergillus spp. Seventy-one patients (age in 1992, 14.1 +/- 7.5 years) were studied. Reactivities for anti-BPI-IgG and IgA were found in 28 (39%) and 26 (37%) patients, respectively. The seroprevalence of anti-BPI-IgA, but not IgG, increased significantly with age. P. aeruginosa colonization was associated with elevated concentrations of anti-BPI-IgG (P = 0.003) and IgA (P = 0.037). There were significant negative correlations between pulmonary function variables (vital capacity, forced expiratory volume in 1 s) in 1992 and 1998, respectively, and concentrations of anti-BPI-IgG or IgA in a multiple regression analysis. Anti-BPI-IgG, but not IgA, remained significantly associated with P. aeruginosa colonization (P = 0.006) and with reduced vital capacity (P = 0.01) in 1998 after correction for total serum isotype concentration. Anti-BPI-IgG are strongly associated with concurrent P. aeruginosa colonization and with long term restrictive pulmonary function abnormalities.

Myocysticercosis: Experience with imaging and therapy

Objective To compare computed tomography (CT) and B-scan ultrasonography (USG) in the diagnosis and to study the efficacy of a combination of oral albendazole and prednisolone in the management of myocysticercosis. Design Retrospective, noncomparative case series. Participants Twenty-six consecutive patients with myocysticercosis. Intervention Diagnostic imaging was performed by CT scan and USG in 24 and 22 patients, respectively; serial USG was obtained in 7 patients receiving treatment. All patients received oral albendazole (15 mg/kg body weight per day) and prednisolone (1.5 mg/kg body weight per day) for 4 weeks. Main outcome measures Presence of scolex on CT scan compared to USG and clinical response to medical therapy were the main outcome measures. Recovery was defined as complete resolution of the scolex or of the main presenting clinical feature. Results Presence of scolex on CT scan (11 of 24) and USG (11 of 22) was not different ( P = 1.0; chi-square test). Recovery was seen in 24 (92%) of 26 patients receiving medical treatment. On serial USG of patients receiving treatment (n = 7), cysts with scolex were seen to progress to a cyst without scolex before final resolution. Time to recovery on treatment (0.5–35 months) correlated with the duration of symptoms at presentation (correlation coefficient r = 0.56, P = 0.003, linear regression analysis), but not with positive serum enzyme-linked immunosorbent assay for anticysticercal antibodies ( P = 0.57, log-rank test) or the presence of scolex ( P = 0.52, log-rank test). Conclusions Treatment with a combination of oral albendazole and prednisolone is effective in the management of myocysticercosis. Imaging methods CT and USG are equally effective in identifying the cyst and the scolex; serial USG is useful in studying the temporal sequence of therapeutic response. The longer recovery time correlating with the duration of symptoms may indicate the chronicity of the inflammatory changes requiring longer time for recovery.

Oral fluid antibody detection in the diagnosis of Helicobacter pylori infection.

The aim of this study was to evaluate an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-Helicobacter pylori specific IgG antibodies in specimens of oral fluid. Antral biopsy specimens, serum and oral fluid samples were collected from 81 patients attending for upper gastrointestinal endoscopy. The presence or absence of current H. pylori infection was determined by culture, histology and urease detection. Anti-H. Pylori specific IgG was detected in serum by an established in-house ELISA and in oral fluid by an ELISA developed for this study. In all, 34 (42%) of 81 patients were positive for H. pylori by one or more of the 'gold standard' tests (culture, histology and urease detection). The oral fluid ELISA had a sensitivity of 94% and specificity of 85% with regard to current H. pylori infection. The serum ELISA had a sensitivity and specificity of 91%. There was an overall agreement of 88% between serum and oral fluid antibody detection. The detection of anti-H. pylori specific IgG in oral fluid by ELISA is comparable in sensitivity and specificity with serum-based methods. Oral fluid-based ELISA could provide a reliable, non-invasive method for the diagnosis of H. pylori infection, and may be of particular benefit for population surveys.

Complementary peptides against the major epitope in the NC16A domain of BP180 show no specificity as vaccines to bullous pemphigoid

A stretch of 14 amino acids (542–555) (MCW-1) in the NC16A domain of BP180 has been shown to be an immunogenic and pathogenic epitope for bullous pemphigoid (BP). Therefore, it provides an excellent target for treatment through a complementary peptide approach, which has been established in other autoimmune diseases, including experimental autoimmune myasthenia gravis. We examined two synthetic complementary peptides BP3CP5 and BP5CP3 against this region. These peptides were derived, respectively, by reading the antisense RNA of this region of BP180 in 3′–5′ and 5′–3′ directions. We found evident complementarities in hydropathic scores between MCW-1 and both complementary peptides. However, by enzyme-linked immunosorbent assay (ELISA), the complementary peptides BP3CP5 and BP5CP3 did not bind to either synthetic peptide BPNP or glutathione- S-transferase (GST) fusion proteins BP180NC16a and GST-BP-1050. BPNP, BP180NC16a and GST-BP-1050 cover the MCW-1 region of BP180 and were used as the natural peptides in this study. In addition, neither BP3CP5 nor BP5CP3 blocked the reaction between BPNP and anti-BPNP antibody, nor did they block immunofluorescent staining of the basement membrane zone by BP sera. Pre-incubation with BP3CP5 and BP5CP3 did not block the binding of BP sera to the BP180NC16a fusion protein in immunoblotting. Furthermore, rabbit antisera raised against BP3CP5 and BP5CP3 did not bind BP sera in ELISA. Pre-incubation with these rabbit antisera did not inhibit or reduce the binding of BP sera to the autoantigen in either immunoblotting or immunofluorescence. Thus, we concluded that complementary peptides against this particular epitope in BP180 NC16A domain showed no specificity as vaccines to BP, although this approach should be tried for other epitopes in various autoimmune bullous diseases.

Vascular endothelial growth factor measured in platelet poor plasma allows optimal separation between cancer patients and volunteers: A key to study an angiogenic marker in vivo?

Serum VEGF levels are elevated in cancer patients and are used as a tumor marker in different malignancies. We have measured VEGF levels in different blood compartments in cancer patients and healthy volunteers in order to assess the most suitable way of processing blood for measuring VEGF as a marker of tumor-angiogenesis. VEGF concentrations were analyzed by an enzyme-linked immunosorbent assay in serum (VEGFS), EDTA plasma (VEGFEDTA), citrated plasma (VEGFC), CTAD-plasma (VEGFCTAD), platelet poor plasma (VEGFPPP), platelet rich plasma after induction of platelet activation (VEGFPRP). Platelet activation was assessed by measuring PF4 concentrations in different plasma samples. We observed higher VEGFS (P = 0.0027), VEGFEDTA (P = 0.003) and VEGFPPP (P = 0.0007) levels in cancer patients than in volunteers; VEGFPRP concentrations showed no significant difference (P = 0.208). Analysis of the correlation between VEGFplt and VEGFS in cancer patients showed a similar correlation in a comparable VEGFS concentration range as in the volunteers. When comparing VEGFC to VEGFCTAD, we find significantly higher VEGF and PF4 levels in citrated plasma (VEGF: P = 0.00019; PF4: P = 0.00023). It is likely that VEGFS in cancer patients encompass platelet-delivered VEGF and VEGF from other sources, notably from (neo)-angiogenesis in tumoral tissue. The best discrimination between volunteers and cancer patients was observed in PPP. As generating plasma can induce platelet activation, with consequent VEGF release from platelets, we suggest that to assess free circulating VEGF, CTAD plasma should be used.

Comparison of serologic tests with urinary antigen detection for diagnosis of legionnaires' disease in patients with community-acquired pneumonia.

OBJECTIVE: To compare serologic methods and detection of urinary antigen in the diagnosis of community-acquired pneumonia. METHODS: Paired sera from 84 patients with community-acquired pneumonia were tested for Legionella pneumophila serogroup (LP SG) 1-7 and Legionella micdadei by use of the indirect immunofluorescence antibody test (IIF), enzyme-linked immunosorbent assay (ELISA) for LP SG 1-7 and complement-fixation (CF) test for LP SG 1. All patients were evaluated by ELISA urinary antigen detection for LP SG 1. RESULTS: Seven patients met the CDC criteria for acute Legionella infection, while in the rest of them we failed to detect urinary Legionella antigen. Thirty-three patients had non-diagnostic IIF antibody titers. Serum ELISA (IgG and/or IgM) was positive in 40 patients. Nine patients showed at least one CF titer of >/=1:32. The sensitivities of ELISA IgM for the first and the second serum samples compared with IIF were 42.8% and 46.6%, respectively, while the specificities were higher, i.e. 87% and 88.4%, respectively. The sensitivities of ELISA IgG for the first and the second samples were 42.8% and 53.3%, and the specificities were 77.9% and 76.8%, respectively. CONCLUSIONS: Although ELISA is simple to perform and easy to automate, we think that its advantages over indirect immunofluorescence and urinary antigen detection remain questionable.

Transport of proteins across the human placenta.

The transport of various proteins across the human placenta was investigated by comparing maternal and fetal concentrations of tetanus antigen (TT-AG), anti-tetanus (TT)-immunoglobulin G (IgG) (following maternal vaccination), IgA, human chorionic gonadotropin (hCG), human placental lactogen (hPL), and alpha-fetoprotein (AFP) at term. The concentrations of the six proteins were determined using enzyme-linked immunosorbent assay in serum of maternal venous and umbilical (fetal) vein samples obtained at delivery from uncomplicated term pregnancies (n = 16). The ratios (mean +/- standard deviation) of fetal (umbilical) to maternal level were 1.41 +/- 0.33 (anti-TT-IgG), 0.91 +/- 0.37 (TT-AG), 0.002 +/- 0.001 (IgA), 0.003 +/- 0.001 (hCG), and 0.008 +/- 0.004 (hPL), while the maternal:fetal concentration ratio of AFP was 0.002 +/- 0.002. IgA, hCG, hPL, and AFP showed a close correlation between maternal and fetal levels varying between r2 = 0.47 to 0.73 (P < 0.004-0.0001). Because AFP is produced by the fetus while IgA originates in the mother, the appearance of small amounts of these two proteins in the maternal or fetal compartment, respectively, suggests a slow rate of diffusion following a high concentration gradient. The detection of hCG and hPL in fetal serum is also interpreted as diffusion from the maternal into the fetal blood. Anti-TT-IgG has a significantly higher concentration in the fetal as compared with the maternal serum, which is in line with the well-documented active transfer of IgG. Fetal TT-antigen levels were similar to maternal concentrations, showing a close correlation (r2 = 0.74, P < 0.0001) between the two proteins. The correlation between maternal and fetal concentrations of various proteins like IgA (150,000 Da), hCG (42,000 Da), and hPL (21,000 Da) suggests passive diffusion of these macromolecules across the placenta from the maternal to the fetal side, albeit at a slow rate. A similar process is postulated for AFP (70,000 Da) diffusing in the opposite direction from the fetus to the mother. There was no significant difference between the transplacental fetomaternal gradient of IgA and hCG and the maternal-fetal gradient of AFP. In view of the substantially larger volume of circulating maternal as compared with fetal blood, a significantly higher rate of crossing of AFP as compared with the other proteins must be assumed. It is uncertain whether a difference in the rate of transplacental transfer in the two directions or an additional source of AFP production in the maternal compartment explains the high maternal level. Anti-TT-IgG concentration is significantly higher in fetal than in maternal serum suggesting active transfer from the mother to the fetus. Furthermore, there is considerable transfer of TT-AG and a close correlation of fetal:maternal ratios of anti-TT-IgG (150,000 Da) and TT-AG (150,000 Da) could be an indication for a specific transfer of the antigen antibody complex.

Purification, potency, and efficacy of the botulinum neurotoxin type A binding domain from Pichia pastoris as a recombinant vaccine candidate.

Recombinant botulinum neurotoxin serotype A binding domain [BoNT/A(Hc)], expressed in Pichia pastoris, was developed as a vaccine candidate for preventing botulinum neurotoxin type A (BoNT/A) intoxication. After fermentation and cell disruption, BoNT/A(Hc) was purified by using a three-step chromatographic process consisting of expanded-bed chromatography, Mono S cation-exchange chromatography, and hydrophobic interaction chromatography. Two pools of immunogenic product were separated on the Mono S column and processed individually. Both products were more than 95% pure and indistinguishable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot analysis, and enzyme-linked immunosorbent assay (ELISA). Each protein was assayed for potency in mice at immunogen doses ranging from 2.4 ng to 10 microg, followed by challenge with 1,000 mouse intraperitoneal 50% lethal doses (i.p. LD50) of BoNT/A. The calculated 50% effective dose for both peaks was approximately 0.1 microg/mouse. Peak 1 was evaluated further in a mouse efficacy assay. Mice were injected either once, twice, or three times at five different doses and subsequently challenged with 100,000 mouse i.p. LD50 of BoNT/A. In general, multiple injections protected better than one, with complete or nearly complete protection realized at doses of >/=0.5 microg/mouse. Serum neutralization and ELISA titers were also determined. Tellingly, 82 of 83 mice with antibody titers of >/=1, 600, as measured by ELISA, survived, but only 6 of 42 mice with titers of </=100 survived. This work shows that the purified BoNT/A(Hc) produced was a highly effective immunogen, able to protect against a high challenge dose of neurotoxin.

Positron emission tomography plus serum TA90 immune complex assay for detection of occult metastatic melanoma

Background: Whole-body positron emission tomography (PET) using 18F-fluorodeoxyglucose is very sensitive for detection of metastatic melanoma. We previously reported the accuracy of a serum TA90 enzyme-linked immunosorbent assay (ELISA) in predicting the recurrence of early-stage melanoma. This serum ELISA measures a circulating immune complex composed of TA90 a tumor-associated 90-kD glycoprotein antigen, and its IgG antibody. We hypothesized that using PET examination in conjunction with our serum TA90 ELISA would increase the detection of occult melanoma lesions. Study Design: From November 1, 1993 to December 31, 1995, 87 patients underwent PET examination followed within 6 weeks by serum TA90 ELISA. All patients had undergone complete resection of local, regional, or distant metastatic melanoma and had no clinical evidence of disease at the time of PET examination. Each patient had complete followup for 12 months after PET examination. The clinical course was determined by chart review of clinical and radiographic findings. Results: Of the 25 patients who experienced recurrence of disease within 6 months of followup, 22 (88%) had a positive result on one or both tests. Of the 52 patients with a negative result on both tests, 49 (94%) remained disease-free at 6 months of followup. Of the 14 patients who developed distant metastases within 6 months after PET examination, 13 (93%) had a positive PET examination and/or a positive TA90 ELISA. Conclusions: The TA90 ELISA correlated well with PET examination for detection of metastatic melanoma. The high sensitivity of PET examination plus TA90 ELISA suggests that this combination may have a role in the surveillance of patients at high risk of developing metastatic melanoma.

Evaluation of North American antibody detection tests for diagnosis of brucellosis in goats.

The sensitivities and specificities of 17 antibody detection tests for brucellosis in goats were estimated. Tests evaluated included the U.S. Department of Agriculture (USDA) card test with 8% cell concentration (8%Card), USDA rapid automated presumptive test (RAP), Mexican rose bengal plate tests with 8 and 3% cell concentrations (8%RB and 3%RB), French rose bengal plate test with 4.5% cell concentration (4.5%RB), USDA standard plate test (SPT), USDA buffered acidified plate agglutination test (BAPA), USDA and Mexican rivanol tests (URIV and MRIV), USDA standard tube tests with Brucella abortus and Brucella melitensis antigens (SATA and SATM), serum enzyme-linked immunosorbent assay (ELISA), USDA cold-fixation complement fixation tests with B. abortus and B. melitensis antigens (CFA and CFM), USDA and Mexican milk ring tests (UBRT and MBRT), and a milk ELISA. Test sensitivity was evaluated by using two groups of 10 goats experimentally infected with B. melitensis or B. abortus and monitored for 24 weeks. Specificity was evaluated by using 200 brucellosis-free nonvaccinated goats from 10 California herds. The 3%RB was considered a good screening test because of high sensitivity at week 24 postinfection (90%), ease of performance, and low cost. The cold-fixation CFA and CFM had 100% specificity in the field study and were considered appropriate confirmatory tests. The milk ELISA was significantly more sensitive (P < 0.05) than the UBRT and significantly more specific (P < 0.05) than the MBRT. The milk ELISA also had the advantage of objectivity and ease of interpretation.

Serum levels of soluble Fas/APO-1 receptor are increased in systemic sclerosis.

It has been suggested that rheumatic diseases may result from a deficit in Fas-mediated T-cell apoptosis. Recent studies have demonstrated increased soluble Fas in sera from lupus erythematosus patients. We were interested to determine whether elevated soluble Fas levels are associated with systemic sclerosis. Soluble Fas levels were retrospectively assayed using a sandwich enzyme-linked immunosorbent assay in serum from 30 patients with systemic sclerosis and 15 normal controls. Hospital medical records were retrospectively reviewed for clinical and laboratory characteristics of the patients. Soluble Fas levels were analysed in subsets of patients with limited (lcSSc) versus diffuse cutaneous systemic sclerosis (dcSSc) and correlated with inflammatory activity. In systemic sclerosis soluble Fas serum levels (lcSSc, 2.19 +/- 0.71 ng/ml, dcSSc 2.53 +/- 1.37 ng/ml) were significantly higher than in normal controls (1.26 +/- 0.36 ng/ml). However, there were no significant differences in soluble Fas levels between lcSSc and dcSSc and poor correlation between soluble Fas levels and inflammatory activity status. Detection of elevated soluble Fas might serve as a clinical marker for immunological dysregulation in systemic sclerosis, but not for inflammatory disease activity.

Use of muscle fluid as a source of antibodies for serologic detection of Salmonella infection in slaughter pig herds.

Fluid drained from a muscle tissue sample was used as an alternative to serum for the detection of specific anti-Salmonella antibodies in an indirect LPS enzyme-linked immunosorbent assay (ELISA). In the first study, serum and muscle fluid from 3 pigs experimentally infected with Salmonella typhimurium showed parallel dilution-response relationships when ELISA optical density (OD) values were plotted against sample dilution. ELISA results obtained with serum diluted 1:400 corresponded to those from muscle fluid diluted 1:30. In a second study, using the predetermined dilutions of individually paired serum and muscle fluid samples from 103 pigs, a high degree of concordance between the serum ELISA and the muscle fluid ELISA was observed. Limits of agreement between the 2 methods were calculated as -8.9 to 12.3 OD%, which was considered acceptable. The muscle fluid ELISA had specificities of 0.91-1.0 and sensitivities of 0.80-0.89 at various cutoff values as compared with the serum ELISA. Muscle fluid is a useful postmortem alternative to serum when used with an ELISA to detect anti-Salmonella antibodies.

Persistent parvovirus B19 infections in immunocompromised children.

Immunocompromised patients have been shown to suffer from prolonged viral infections often without detectable immune response. Here chronic infections with low virus levels can be frequently observed. In these patients viral DNA can be detected over long periods by polymerase chain reaction (PCR). In this study parvovirus B19 presence was assessed by PCR, immunoblot and enzyme-linked immunosorbent assay in sera from children with mainly oncological and hematological diseases. In 45% of sera B19 DNA was observed. Of the children 25% had IgG antibodies to viral protein 1 and 2 (VP1/2) and 15% to nonstructural protein 1 (NS1). In 6% of children IgM antibodies to VP1/2 were detected. These results indicate that the number of children with immune response to B19 proteins is distinctly lower than the number of children with B19 DNA. Transfusions of blood products might have been a possible route for B19 infection. Establishment and maintenance of a persistent parvovirus B19 infection with or without immune response are enhanced in the analyzed immunocompromised children in comparison with immunocompetent children. A persistence of B19 DNA was demonstrated up to 10 months in patients sera.