IgG Enzyme-linked Immunosorbent Assay Research Articles

Antidesmoglein 1 and 3 antibodies in healthy subjects of a population in the Peruvian high amazon.

The objective of this study was to determine the presence of anti-Dsg1 and Dsg3 antibodies in healthy subjects of the high Peruvian Amazon (Tuemal, Rodriguez de Mendoza province, department of Amazonas) to establish the theoretical presence of environmental factors or triggers in the area. Cross-sectional study. The study population included persons of any age or gender, clinically healthy, who were evaluated by a dermatologist to confirm the absence of blistering diseases. Blood samples were analyzed by indirect immunofluorescence (IIF), immunoprecipitation (IP), anti-Dsg1 IgM antibody (Ab) enzyme-linked immunosorbent assay (ELISA), as well as anti-Dsg1 and anti-Dsg3 IgG Ab ELISA. Participants included 21 healthy subjects comprised of 61.9% males and 38.1% females; 47.6% had a positive anti-Dsg1 Ab ELISA for total IgG (or any subclasses). IIF detected antibodies against intercellular spaces in one subject. Anti-Dsg1 Ab IP was mildly positive in 33.3% of the subjects. Anti-Dsg1 IgG subclasses found positive were: IgG1 (19.0%), IgG2 (33.3%), and IgG3 (28.6%); none of the samples were positive for anti-Dsg1 Ab IgM ELISA, and 23.8% of the subjects were positive for anti-Dsg3 Ab ELISA. The age distribution was similar for subjects positive for anti-Dsg1 and anti-Dsg3 Ab ELISA, with higher frequencies found among the 20-29 and 40-49year-old age groups. A fraction of healthy subjects of the high Peruvian Amazon developed anti-Dsg1 and anti-Dsg3 antibodies, demonstrating the possible presence of environmental factors for endemic pemphigus (EP) at a higher altitude than previously described.

Optimized enzyme-linked immunosorbent assay for detecting cytomegalovirus infections during clinical trials of recombinant vaccines

BackgroundIn clinical trials of cytomegalovirus (CMV) glycoprotein B (gB) vaccines, CMV infection is detected by first depleting serum of anti-gB antibodies and then measuring anti-CMV antibodies with a commercially available enzyme-linked immunosorbent assay (ELISA) kit, with confirmation of positive findings by immunoblot. ObjectivesIdentification of CMV immunoantigens for the development of an ELISA that detects specifically CMV infection in clinical samples from individuals immunized with gB vaccines. Study designSensitivity and specificity of ELISAs using antigenic regions of CMV proteins UL83/pp65, UL99/pp28, UL44/pp52, UL80a/pp38, UL57, and UL32/pp150 were measured. ResultsAn IgG ELISA using a UL32/pp150 [862–1048] capture peptide was the most specific (93.7%) and sensitive (96.4%) for detecting CMV-specific antibodies in sera. The ELISA successfully detected CMV-specific antibodies in 22 of 22 sera of subjects who had been vaccinated with a gB vaccine but who had later been infected with CMV. The ELISA was linear over a wide range of CMV concentrations (57–16,814 ELISA units/mL) and was reproducible as indicated by a 5% intra-day and 7% inter-day coefficients of variation. The signal was specifically competed by UL32/pp150 [862–1048] peptide but not by CMV-gB or herpes simplex virus 2 glycoprotein D. Lipid and hemoglobin matrix did not interfere with the assay. ConclusionThe UL32/pp150 [862–1048] IgG ELISA can be used for the sensitive and specific detection of CMV infection in gB-vaccinated individuals.

Missed opportunities for the diagnosis of Brucella infection among slaughterhouse workers at the Kumasi Abattoir, Ghana

Brucellosis is a zoonotic disease in humans with its public health importance. Laboratory diagnostic methods targeting brucellosis are not performed in hospital settings across Ghana. Very little is known about the comparative diagnostic abilities of the various tests available presently. The aim of this study therefore was to evaluate and compare diagnostic performances of Rose Bengal Plate Test (RBPT), Enzyme Linked Immunosorbent Assay (ELISA) and Polymerase Chain Reaction (PCR) employed in diagnosing Brucella infection.Two hundred and twenty Abattoir workers were randomly selected in Kumasi, Ghana. Blood samples were collected, serum extracted and tested for the presence of anti-Brucella antibodies and compared among three different techniques, using ELISA, RBPT and PCR.From the 220 participants tested for antibodies against Brucella spp., 3 (1.4%), 4 (1.8%) and 21 (9.6%) were positive for Rose Bengal Plate test, anti-Brucella ELISA IgM and anti-Brucella ELISA IgG respectively. A total of 98 (44.5%) participants tested positive by PCR. The sensitivity, specificity, positive predictive value, negative predictive values and Kappa value for Rose Bengal in comparison with PCR were 66.7%, 55.8%, 2.0%, 100% and 0.013 respectively while that for ELISA IgG in comparison with PCR were 85.7%, 71.3%, 18.4%, 98.5% and 0.212 respectively.PCR yielded the highest sensitivity and specificity among the three diagnostic methods in this study and should be considered for use at strategic reference laboratories to augment existing routine serological tests for brucella performed in laboratories in Ghana.

Risk Factors Associated With Bordetella pertussis Among Infants ≤4 Months of Age in the Pre-Tdap Era: United States, 2002-2005.

In the United States, infants have the highest reported pertussis incidence and death rates. Improved understanding of infant risk factors is needed to optimize prevention strategies. We prospectively enrolled infants ≤4 months of age with incident-confirmed pertussis from 4 sites during 2002-2005 (preceding pertussis antigen-containing vaccination recommendations for adolescents/adults); each case-patient was age and site matched with 2 control subjects. Caregivers completed structured interviews. Infants and their contacts ≥11 years of age were offered serologic testing for IgG; being seropositive was defined as ≥94 antipertussis toxin IgG enzyme-linked immunosorbent assay units per milliliter. Enrolled subjects (115 case-patients; 230 control subjects) had 4396 contacts during incubation periods; 83 (72%) case-patients had ≥1 contact with prolonged (≥5 days) new cough in primary or secondary households. In multivariable analysis, the odds for pertussis were higher for infants with primary/secondary household contacts who had a prolonged new cough, compared with infants who did not. These contacts included mother [adjusted matched odds ratio (aMOR), 43.8; 95% confidence interval (CI), 6.45-298.0] and ≥1 nonmother contact (aMOR, 20.1; 95% CI, 6.48-62.7). Infants receiving breast milk with 0-1 formula feedings daily had decreased pertussis odds (aMOR, 0.27; 95% CI, 0.08-0.89), compared with those receiving more formula. Of 41 tested case-patients, 37 (90%) were seropositive. Pertussis in infants was associated with prolonged new cough (≥5 days) in infants' household contacts. Findings suggest that breastfeeding protects against pertussis and warrants recommendation with pertussis prevention strategies, which currently include pertussis vaccination of pregnant mothers and infants' close contacts.

Serological Survey of Hydatid Disease in Asymptomatic Peoples in Mosul City, Iraq

Hydatidosis is one of the major zoonotic diseases that causes considerable economic losses and public health problems worldwide. This study was conducted in order to determine the seropositivity of hydatid disease in apparently healthy individuals who are attending the main blood bank in Mosul City. Therefore, enzyme linked immunosorbent assay (ELISA) was used in this study and to find a relation between the seropositivity and age, gender, occupation and educational level and to monitor the impact of the control programs for the disease in Mousl City. The study was conducted during 9 months period extended from November 2012 to July 2013.The samples of the study were composed of 480 apparently healthy individuals with ages ranging between 18-60 years, 280 male and 200 female. Serum samples were obtained from the main blood bank in Mosul city, ELISA test was used for detecting specific anti Echinococcus granulosus antibodies in patients’ sera. ELISA kits used were (Echinococcus ELISA IgG- IBL, Hamburg, Germany –RE 56201) kits. The results revealed that out of 480 cases only 24 (5%) were positive by ELISA, the most frequently infected individuals were females (8%) while males were less frequently encounter the disease (2.86 %). the rate of hydatid cysts was highest at age group 21-30 years (7.02%) followed by age group 31-40 years (6.5%), 41-50 years ( 3.7% ) respectively, the highest infection rate was reported among housewives (10.83%), followed by self employed (4.46%), then the government employed (3.23%). On the other hand ,the lowest rate of infection was recorded among students (1.28%) and (2.88%) in other occupations, it was reported that (56.5%) were illiterate, (4.35%) were primary school, (2.42%) were secondary school and (1.38%) were of high education, most of the seropositive individuals were from rural or semi-rural areas surrounding Mosul City. It was recorded that (6.02%) and ( 3.89% ) were from rural and urban area respectively. The rate of seropositivity in the individuals who have contact with dogs (5.81%) was higher than that in individuals who do not have contact with dogs (4.05%).

ІМУНОЛОГІЧНИЙ МОНІТОРИНГ ДОРОСЛОГО НАСЕЛЕННЯ ЛЬВІВЩИНИ ЩОДО КАШЛЮКУ У 2016 РОЦІ

Мета – провести серологічний моніторинг серед дорослого населення щодо кількісного визначення імуноглобулінів класу G до кашлюку.Матеріали і методи. Дослідження проводились з використанням методу імуноферментного аналізу.Результати досліджень та їх обговорення. Серонегативний прошарок населення становить 50,0 %. Специфічні імуноглобуліни G у високих титрах (понад 50 BU/ml) були виявлені у 3,75 % осіб, що свідчить про період реконвалесценції після кашлюку.Висновки. Використання цього методу підтверджує наявність випадків захворювання на кашлюк серед дорослого населення при недостатній настороженості медичних працівників щодо захворюваності на кашлюк даної вікової групи. Підтверджено об’єктивність і інформативність методу та доцільність його широкого впровадження в практику громадського здоров’я з метою здійснення належного епідеміологічного нагляду.

An ELISA assay using a combination of recombinant proteins from multiple strains of Orientia tsutsugamushi offers an accurate diagnosis for scrub typhus

BackgroundScrub typhus (ST) is a disease caused by an obligate intracellular bacterium, Orientia tsutsugamushi, an organism that requires a BSL3 laboratory for propagation. The disease is hallmarked by an eschar at the site of the chigger bite, followed by the development of fever, malaise, myalgia, anorexia, and papulomacular rash. Indirect immunofluorescent assay (IFA) is the gold standard for scrub typhus diagnosis, however, the subjectivity of the assay, the need for a specialized laboratory and instruments has limited the wide use of the test in resource limited areas.MethodsA recombinant-protein based enzyme linked immunosorbent assay (ELISA) using the most abundant and immunodominant protein for the detection of Orientia specific antibodies in serum has been developed. The performance of the assay was evaluated using prospectively collected acute sera from 248 randomly selected patients in Thailand. The ELISA assay was evaluated using two different cutoff values.ResultsThe receiver operating characteristic (ROC) curve generated cutoff values gave slightly better consistency with diagnosis of ST than those cutoff values established by averaging ELISA optical density of known negatives at 99% confidence interval. Both cutoff values provided similar statistical parameters when compared with the diagnosis of ST, indicating the validity of both calculations to derive cutoff values. These results suggest that both IgG and IgM ELISA performed well to accurately diagnose scrub typhus cases in endemic areas using only acute serum samples.ConclusionsWe have successfully developed an ELISA assay for the detection of Orientia-specific antibodies in serum that could provide effective screening of acute sera under clinical setup and it is also a useful assay to estimate seroprevalence in various endemic areas.

An ELISA for the determination of human IgG based on the formation of a colored iron(II) complex and photometric or visual read-out

The paper describes a colorimetric sandwich enzyme-linked immunosorbent assay (ELISA) for the determination of human IgG. It is based on the use of an Fe(II) coordination complex as a signal amplifier and of mesoporous silica nanoparticles modified with glucose oxidase (GOx) and secondary antibodies (Ab2). After formation of the immuno sandwich complex, the quantity of GOx is proportional to the quantity of IgG. On addition of Fe(II) and glucose, GOx catalyzes the oxidation of glucose to produce hydrogen peroxide which oxidizes Fe(II) to Fe(III). After adding a stop solution containing the complexing ligand 1,10-phenanthroline (Phen), un-reacted Fe(II) forms an orange-red complex with Phen which can be detected by plate reader and even seen with bare eyes. This sandwich ELISA has a linear response in the 1 pg.mL−1 to 100 ng.mL−1 human IgG concentration range and a 860 fg.mL−1 detection limit. This is 20 times lower than the commercial ELISA for human IgG. The assay also is selective, stable, highly sensitive and cost-effective.

Detection of IgE autoantibodies to BP180 and BP230 and their relationship to clinical features in bullous pemphigoid

IgE autoantibodies are considered to be involved in the pathogenesis of bullous pemphigoid (BP), particularly inflammatory and erythematous phenotypes. To develop reliable enzyme-linked immunosorbent assays (ELISAs) for the detection of IgE autoantibodies to both BP180 and BP230 in BP sera, and to compare the ELISA results with clinical features. We used commercially available IgG ELISAs to develop IgE ELISAs for both BP180 and BP230. To determine the influence of excess amounts of IgG autoantibodies, all normal and BP sera were tested before and after IgG adsorption. The results of the IgE ELISAs were statistically compared with various ELISAs and various clinical parameters, including our own severity scores and BP phenotypes. IgG adsorption generally showed no changes in sensitivity and specificity for IgE ELISAs, although slight cross-reactivity of anti-IgE secondary antibody to IgG and interference of excess amounts of IgG autoantibodies to IgE reactivity were suggested. IgE autoantibodies to BP180 were found in 21 of 36 BP sera and IgE autoantibodies to BP230 were found in 18 of 36 BP sera. The results of IgG and IgE ELISAs for both BP180 and BP230 were well correlated. IgG and IgE anti-BP180 antibodies correlated with disease activity but IgG and IgE anti-BP230 autoantibodies did not. IgE anti-BP230 autoantibodies correlated with nodular phenotype but not erythematous phenotype. The results of this study indicated that IgE autoantibodies to both BP180 and BP230 are frequently detected in BP sera. IgE anti-BP180 autoantibodies seemed to be pathogenic, while an association between IgE autoantibodies and inflammatory BP phenotype was not indicated.

A clinical and serological study of linear IgA bullous dermatosis without linear immunoglobulin deposition other than IgA at the basement membrane zone using direct immunofluorescence.

Linear IgA bullous dermatosis (LABD) is a heterogeneous disease. Different diagnostic criteria have been used in different reports. To reappraise the characteristic features of LABD with only IgA deposition at the basement membrane zone (BMZ) with direct immunofluorescence (DIF). We retrospectively collected data from 101 patients from our archival records from 1 January 1996 to 31 December 2014 who had: (i) blisters on the skin and/or mucous membranes; (ii) subepidermal blisters in a biopsy specimen; and (iii) linear IgA deposition along the BMZ with/without linear C3 deposition at the BMZ with DIF. Most patients were referred for serological evaluation. Patients who showed concurrent linear IgG and/or IgM deposition at the BMZ under DIF were excluded. Clinical manifestations and serological findings were analysed. Heterogeneity of autoantigens in LABD was confirmed. Fifty-four of 101 patients (53%) had IgG antibodies detected either by indirect immunofluorescence, immunoblotting or enzyme-linked immunosorbent assays (ELISA). No statistical difference in clinical manifestations was found between patients in the IgG antibody-possessing group and patients in the other group. An association of IgG anti-BMZ antibodies with LABD may increase if new IgG immunoblotting or ELISA techniques become available. Consensus for diagnostic criteria for LABD is desired for prospective data storage, although it may be arbitrary.

Comparison of in-house biotin-avidin tetanus IgG enzyme-linked-immunosorbent assay (ELISA) with gold standard in vivo mouse neutralization test for the detection of low level antibodies

BackgroundDetection of anti-tetanus antibody levels is necessary for both determination of the immune status of individuals and also for planning preventive measures. ELISA is the preferred test among in vitro tests however it can be affected by the cross reacting antibodies. A previously developed in-house ELISA test was found not reliable for the antibody levels ≤1.0IU/ml. A new method was developed to detect low antibody levels correctly. The aim of the present study was to compare the results of the newly developed in-house biotin-avidin tetanus IgG ELISA test with the in vivo mouse neutralization test, for the antibody levels ≤1.0IU/ml. MethodsA total of 54 serum samples with the antibody levels of three different levels, =0.01IU/ml, 0.01–0.1IU/ml, 0.1–1IU/ml, which were detected by in vivo mouse neutralization test were studied by the newly developed in-house biotin-avidin tetanus IgG ELISA test. Test was validated by using five different concentrations (0.01IU/ml, 0.06IU/ml, 0.2IU/ml, 0.5IU/ml, 1.0IU/ml). ResultsA statistically significant correlation (r2=0.9967 p=0,001) between in vivo mouse neutralization test and in-house biotin-avidin tetanus IgG ELISA test, was observed. For the tested concentrations intra-assay, inter-assay, accuracy, sensitivity, specificity and coefficients of variations were determined as ≤15%. ConclusionIn-house biotin-avidin tetanus IgG ELISA test can be an alternative method to in vivo mouse neutralization method for the detection of levels ≤1.0IU/ml. By using in-house biotin-avidin tetanus IgG ELISA test, individuals with non protective levels, will be reliably detected.

Association between seroprevalence of measles and various social determinants in the year following a measles outbreak in Turkey

ObjectivesDespite an ongoing measles elimination programme, a measles outbreak occurred in 2013 in Turkey. Population-based seroprevalence studies are needed to determine seronegativity and explore the reasons for this outbreak. This study aimed to explore the seroprevalence of measles and its association with various social determinants in a provincial population in Turkey in the year following a measles outbreak. Study designCross-sectional study. MethodsThis study was conducted in Manisa Province in 2014 in a sample of 1740 people aged >2 years. The dependent variable was the seroprevalence of measles. Independent variables were sex, age, migration, household size, household density, income, education level, existence of chronic disease and occupational class. Blood samples were collected from participants at family health centres. The presence of specific measles antibodies in serum samples was determined using an anti-measles virus IgG enzyme-linked immunosorbent assay test. Chi-squared test and logistic regression analysis were performed. ResultsOverall, data from 1250 people were analysed. The seroprevalence of measles in the whole study population was 82.2% (95% confidence interval 80.0–84.2). Seroprevalence was 55.4% among subjects aged 2–9 years, 48.7% among subjects aged 10–19 years, 74.1% among subjects aged 20–29 years and 93.6% among subjects aged 30–39 years (P < 0.01). Seroprevalence in subjects aged >40 years was >95%. The lowest seroprevalence was found in primary school children (40.2%), followed by those below the age for primary education (69.8%) and secondary school graduates (75.1%). The prevalence of measles seronegativity was not associated with any of the social determinants when adjusted for age. ConclusionsThe seroprevalence of measles was lower than expected in the study population and was particularly low in subjects aged <30 years of age despite previous vaccination. Seroprevalence was not associated with social determinants of health that confirmed either an even distribution of virus exposure or fair access to vaccination services. However, the current seroprevalence cannot be sufficiently effective to reach the measles elimination targets, suggesting that it may be necessary to re-evaluate the need for an extra dose of measles vaccine.

Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA).

The enzyme-linked immunosorbent assay (ELISA) technique is based on the specific recognition ability of the molecular structure of an antigen (epitope) by an antibody and is likely the most important diagnostic technique used today in bioscience. With this methodology, it is possible to diagnose illness, allergies, alimentary fraud, and even to detect small molecules such as toxins, pesticides, heavy metals, etc. For this reason, any procedures that improve the detection limit, sensitivity or reduce the analysis time could have an important impact in several fields. In this respect, many methods have been developed for improving the technique, ranging from fluorescence substrates to methods for increasing the number of enzyme molecules involved in the detection such as the biotin–streptavidin method. In this context, nanotechnology has offered a significant number of proposed solutions, mainly based on the functionalization of nanoparticles from gold to carbon which could be used as antibody carriers as well as reporter enzymes like peroxidase. However, few works have focused on the study of best practices for nanoparticle functionalization for ELISA enhancement. In this work, we use 20 nm gold nanoparticles (AuNPs) as a vehicle for secondary antibodies and peroxidase (HRP). The design of experiments technique (DOE) and four different methods for biomolecule loading were compared using a rabbit IgG/goat anti-rabbit IgG ELISA model (adsorption, directional, covalent and a combination thereof). As a result, AuNP probes prepared by direct adsorption were the most effective method. AuNPs probes were then used to detect gliadin, one of the main components of wheat gluten, the protein composite that causes celiac disease. With this optimized approach, our data showed a sensitivity increase of at least five times and a lower detection limit with respect to a standard ELISA of at least three times. Additionally, the assay time was remarkably decreased.

Seroprevalence and risk factors for infection with equine coronavirus in healthy horses in the USA

Equine coronavirus (ECoV) is considered an enteric pathogen of foals and has only recently been associated with infections in adult horses. Seroprevalence data is needed to better understand the epidemiology of ECoV in adult horses, evaluate diagnostic modalities and develop preventive measures. The objective of this study was to investigate the seroprevalence and selective risk factors for ECoV in 5247 healthy adult horses in the USA, using a recently established and validated IgG enzyme-linked immunosorbent assay. Prevalence factors analysed in this study included geographic region, age, breed, sex and use.A total of 504/5247 horses (9.6%) horses tested seropositive. Geographic region (Mid-West; P = 0.008), breed (Draft horses; P = 0.003) and specific uses of horses (ranch/farm, P = 0.034; breeding use, P = 0.016) were all statistically significant risk factors for seropositivity.

Recombinant LipL32 Protein Developed Using a Synthetic Gene Detects Leptospira-specific Antibodies in Human Serum Samples.

Synthetic biology is emerging as a viable alternative for the production of recombinant antigens for diagnostic applications. It offers a safe alternative for the synthesis of antigenic principles derived from organisms that pose a high biological risk. Here, we describe an enzyme-linked immunosorbent assay (ELISA) using the synthetic recombinant LipL32 (rLipL32) protein expressed in Escherichia coli for the detection of Leptospira-specific antibodies in human serum samples. The rLipL32-based ELISA was compared with a microscopic agglutination test (MAT), which is currently used as the gold standard for the diagnosis of leptospirosis. Our results showed that all the MAT-positive serum samples were positive for Leptospira-specific IgG in an ELISA, while 65% (n = 13) of these samples were also positive for Leptospira-specific IgM. In the MAT-negative serum samples, 80% and 55% of the samples were detected as negative by an ELISA for Leptospira-specific IgM and IgG, respectively. An ELISA using the synthetic rLipL32 antigen was able to distinguish Leptospira-specific IgM (sensitivity 65% and specificity 80%) and IgG (sensitivity 100% and specificity 55%) in human serum samples and has the potential to serve as a rapid diagnostic test for leptospirosis.

ELISA Methods for the Detection of Ebolavirus Infection.

Ebola viruses are high-priority pathogens first discovered in rural Africa associated with sporadic outbreaks of severe hemorrhagic disease in humans and nonhuman primates. Little is known about the disease ecology or the prevalence of past exposure of human populations to any of the five species of the genus Ebolavirus. The use of immunologic means of detection for either virus antigens or the host's immune response to antigen associated with prior infections offers a powerful approach at understanding the epidemiology and epizootiology of these agents. Here we describe methods for preparing antigen detection sandwich enzyme-linked immunosorbent assays (ELISAs) as well as IgG and IgM ELISAs for the detection of ebolavirus antigens or antibodies in biological samples.

Echinococcosis: Unexpected Occurrence and the Diagnostic Contribution of Routine Histopathology.

Echinococcus is the smallest of the major tapeworms. Humans are incidental hosts who become infected by exposure to soil contaminated with ova from the feces of the canine definitive hosts. The major species are E. granulosus (cystic echinococcosis) and E. multilocularis (alveolar or sylvatic echinococcosis), both common worldwide but unusual and not reportable in the United States. Human disease is characterized by slowly growing visceral cystic masses often containing daughter cysts and hydatid sand. In nonendemic areas, the clinical evaluation includes imaging, serology, and surgery; metastatic tumor is a major consideration. The diagnosis depends on the morphologic recognition of scolices and hooklets, albeit degenerated, within the cyst or attached to the cyst wall. Highly suggestive cyst wall features are an acellular hyaline lining surrounded by focally calcific fibrous tissue and chronic inflammation. We report 7 patients encountered in the Chicago area (5 E. granulosus, 2 E. multilocularis) who presented with cystic masses affecting the liver, lung, soft tissue, and spleen. The initial diagnosis was established or suggested by intraoperative evaluation of cyst fluid or cyst wall histopathology. Organism parts were found in 5 cases; 1 patient had a hyalinized cyst wall with a positive IgG (enzyme-linked immunosorbent assay), and 1 had a positive Western blot and a 30-year history of treatment. Treatment included cyst resection, hepatic lobectomy, intraoperative cyst injection, and benzimidazole therapy. There was 1 death. In a nonendemic area, the awareness of the pathologist to the possibility of this disease will exclude a neoplasm and facilitate prompt antihelminthic treatment.

ELISA Serology for Antibodies Against Chlamydia trachomatis in Crohn's Disease.

BackgroundRecently we reported IgA anti-Chlamydia antibodies in patients with Crohn’s disease (CD), in particular in four patients from a single family of six with CD.MethodsWe studied sera from four cohorts from the north of France. These were identified as: EPIMAD (80 pediatric onset CD and 20 pediatric onset ulcerative colitis), MINOTOR (148 adult onset sporadic CD and 50 adult onset ulcerative colitis), Grande Famillies (50) and matched controls for the Grande Famillies cohort (49). Sera were tested using commercial anti-Chlamydia trachomatis (LGV2:434) IgG and IgA human enzyme-linked immunosorbent assay (ELISA) kits. Cutoff for positivity was 11.0 standard units.ResultsPatients with sporadic CD, unaffected first degree relatives from multiplex families and ulcerative colitis patients had no greater serologic reactivity than controls. However, multiplex families’ patients had twice as many positives as the other groups: for IgG 20% vs. 8%; for IgA 20% vs. 10%.ConclusionsThough not attaining statistical significance, the data showed that familial CD patients had greater exposure to C. trachomatis than sporadic CD patients, supporting our earlier results from one family from the north of France. More specific serologic tests based on outer membrane proteins will need to be employed against the various Chlamydia species with zoonotic potential.

Investigation of zoonotic infections in risk groups in Ordu University Hospital, Turkey.

Zoonotic diseases, which are a major public health problem in our city, have a negative impact on public health and also cause economic losses due to yield losses of animals and deaths. This study was carried out to determine the seroprevalence of tularemia, bartonellosis, brucellosis, Q fever, and cystic echinococcosis in the risk groups for zoonotic infection. Ninety serum samples were taken from people in the risk groups in covering veterinarian, butchers, farmers and examined with the following tests: Microagglutination test for tularemia, indirect fluorescent antibody test (IFAT) for bartonellosis, standard tube agglutination test for brucellosis, IFAT IgG for Q fever, and enzyme-linked immunosorbent assay IgG test for cystic hydatid. The Chi-square analysis was used to assess, and the logistic regression analysis was used to identify the risk factors. The analyzed all serum samples were found to be seronegative for tularemia, bartonellosis, and hydatid cyst antibodies. When analyzed for Coxiella burnetii with IgG antibody titers, it was determined that 23 samples (25.6%) were seropositivity. When brucellosis was analyzed with serological tests for Brucella, it was positive in seven samples (7.8%). In this study, examined in the risk groups in which it is located along black sea coast of Turkey for tularemia, bartonellosis, and hydatid cysts, seropositivity was not found. When Brucella was tested, 7.8% was found to be positive, and when analyzed in terms of Q fever, 25.6% of people were determined to be seropositive. In conclusion, in our region, Q fever seropositivity was found to be higher in the risk groups. Therefore, most of the zoonotic disease look like not so common in the region, out of tularemia.

Hepatitis E Virus in Wild Boar in Northwest Poland: Sensitivity of Methods of Detection.

In northwest Poland, 163 blood and 53 fecal samples of wild boars were collected in winter 2012/13 and 2013/14. All blood samples were tested for the presence of hepatitis E virus (HEV) ribonucleic acid (RNA) by two reverse transcription-polymerase chain reaction (RT-PCR) based methods and by anti-HEV IgG enzyme-linked immunosorbent assay (ELISA). About 17.2% of blood samples were seropositive. One-step nested RT-PCR turned out to be too insensitive (11.6% were positive). Therefore a two-step nested RT-PCR was applied where 25.8% of the blood samples were tested positive for HEV RNA. About 50.0% of blood samples positive in ELISA were also positive in two-step nested RT-PCR. The prevalence of HEV RNA in feces was 9.4%. Based on the results of blood (ELISA, PCR) and fecal (PCR) tests, the overall prevalence of HEV in wild boars in northwest Poland was 36.8%. There was no correlation between the ELISA results and the presence of HEV RNA in plasma or in feces. According to the sequencing results of 348 bp PCR products of HEV, there were four different subtypes identified. Reports on the prevalence of HEV in wild boar populations are varying due to different sensitivities of the detection methods. However, this study reveals based on a highly sensitive method that HEV is widely spread in wild boar populations in the northwestern region of Poland and posing a potential risk to the consumer of game meat.