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CD80 expression and infiltrating regulatory T cells in idiopathic nephrotic syndrome of childhood.

CD80 (also known as B7-1) is a costimulatory molecule and was reported to be expressed in biopsies and also excreted in urine from patients with minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS). CD80 is inhibited by CTLA4, which is mainly expressed on regulatory T cells (Tregs). Ineffective circulating Treg response was accused in pathogenesis of nephrotic syndrome. In this study, we aimed to evaluate CD80 expression and infiltrating Tregs in children with MCD and FSGS.

2509 related Products with: CD80 expression and infiltrating regulatory T cells in idiopathic nephrotic syndrome of childhood.

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Development of Multispecies Recombinant Nucleoprotein-Based Indirect ELISA for High-Throughput Screening of Crimean-Congo Hemorrhagic Fever Virus-Specific Antibodies.

Crimean-Congo hemorrhagic fever (CCHF) is a re-emerging zoonotic viral disease prevalent in many parts of Asia, Europe, and Africa. The causative agent, Crimean-Congo hemorrhagic fever orthonairovirus (CCHFV), is transmitted through hard ticks. Tick vectors especially belonging to the species serve as the reservoir and amplifying host. The vertebrate animals including sheep, goat, and bovine act as a short-lasting bridge linking the virus and ticks. CCHFV causes fatal hemorrhagic fever in humans. Humans are usually infected with CCHFV either through the bite of infected ticks or by close contact with infected animals. Immunological assays, primarily enzyme-linked immunosorbent assay (ELISA) using whole viral antigen, are widely used for serosurveillance in animals. However, the whole virus antigen poses a high biohazard risk and can only be produced in biosafety level 4 laboratories. The present study focuses on the development and evaluation of safe, sensitive, and specific IgG indirect enzyme-linked immunosorbent assay (iELISA) using recombinant nucleoprotein (NP) of CCHF virus as an antigen. The codon-optimized NP gene sequence was synthesized, cloned, and expressed in pET28a+ vector. The recombinant NP was purified to homogeneity by affinity chromatography and characterized through Western blot and MALDI-TOF/MS analysis. The characterized protein was used to develop an indirect IgG microplate ELISA using a panel of animal sera. The in-house ELISA was comparatively evaluated vis-à-vis a commercially available ELISA kit (Vector-Best, Russia) with 76 suspected samples that revealed a concordance of 90% with a sensitivity and specificity of 79.4 and 100%, respectively. The precision analysis revealed that the assay is robust and reproducible in different sets of conditions. Further, the assay was used for serosurveillance in ruminants from different regions of India that revealed 18% seropositivity in ruminants, indicating continued circulation of virus in the region. The findings suggest that the developed IgG iELISA employing recombinant NP is a safe and valuable tool for scalable high-throughput screening of CCHFV-specific antibodies in multiple species.

1995 related Products with: Development of Multispecies Recombinant Nucleoprotein-Based Indirect ELISA for High-Throughput Screening of Crimean-Congo Hemorrhagic Fever Virus-Specific Antibodies.

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A new reliable test for crossmatching: microplate hydrogel immunoassay technology.

To provide a novel assay to detect incomplete antibodies in the crossmatching test.

2263 related Products with: A new reliable test for crossmatching: microplate hydrogel immunoassay technology.

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Brucellosis and chlamydiosis seroprevalence in goats at livestock-wildlife interface areas of Zimbabwe.

In Zimbabwe, there have been no chlamydiosis and limited brucellosis studies in goats. This study was conducted to determine the seroprevalence and risk factors of the two diseases in goats at three different livestock-wildlife interface areas: porous, non-porous and non-interface in the south-eastern lowveld of Zimbabwe. Collected sera (n = 563) were tested for Brucella antibodies using the Rose Bengal plate test (RBPT) and the complement fixation test (CFT); and for Chlamydia abortus antibodies using the CFT. All tested goats were negative for Brucella antibodies. Overall, chlamydial seroprevalence was 22%. The porous [c2 = 9.6, odds ratio (OR) = 2.6, p = 0.002] and non-porous (c2 = 37.5, OR = 5.8, p < 0.00001) interfaces were approximately three and six times more likely to be chlamydial seropositive than the non-interface area, respectively. Chlamydial seroprevalence was not associated with sex (c2 = 0.5, OR = 1.2, p = 0.5), abortion history in female goats (c2 = 0.7, OR = 1.3, p = 0.4), keeping goats with cattle (c2 = 0.2, OR = 1.5, p = 0.7) or flock size (c2 = 0.03, OR = 1.4, p = 0.9). Our study provides the first serological evidence of chlamydiosis in goats in Zimbabwe and the results suggest that proximity to wildlife is associated with increased chlamydial seropositivity. Further studies are required to determine the role of chlamydial infection on goat reproductive failure and that of wildlife on C. abortus transmission to domestic ruminants.

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Molecular and seroepidemiological survey on Crimean-Congo Hemorrhagic Fever Virus in Northeast of Iran.

Crimean-Congo hemorrhagic fever (CCHF) is a prevalent tick-borne disease in different regions of Iran. This molecular and serologic study was performed to investigate the Crimean-Congo hemorrhagic fever virus (CCHFV) in collected ticks and in blood samples of some domestic animals in North Khorasan, Northeast of Iran. In this cross sectional study, 136 blood samples from domestic animals (sheep, goats, and cows) collected in the Northeast region in Iran were examined using IgG ELISA assay. Ticks (n = 1478) were collected from sheep, goats, and cows. Out of all collected ticks, 62 specimens were investigated for CCHF virus genome using RT-PCR technique. The data were descriptively presented by median and 95% confidence interval (CI). CCHFV infection rate was 8.1% in studied ticks. Two species of ticks, Hy. anatolicum (n=3; 15%, 95% CI 9.41-20.59) and Rh. sanguineus (n=2; 6.9%, 95%CI 4.33-8.58), were infected with CCHFV genome and were probable vectors of CCHF virus in the area. Infection rate was 15.4% for CCHFV in tested domestic animals. Serologic tests detected CCHFV specific IgG antibodies in 16.2% (95% CI 13.49-18.83) (99/16) and 19.2% (95% CI 13.26-25.20) (26/5) of sheep and goats, respectively. The present study showed that domestic animals and ticks were infected with Crimean-Congo hemorrhagic fever virus and that the disease was endemic in North Khorasan province, Iran. However, further surveillance and prevention programs are recommended.

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Seroepidemiological study of Q fever, brucellosis and tularemia in butchers and slaughterhouses workers in Lorestan, western of Iran.

Most zoonoses are occupational diseases. Q fever, brucellosis and tularemia are major zoonotic diseases for butchers and slaughterhouse workers. However, little information is available about these infectious diseases in such professional populations in western of Iran. The aim of this study was to investigate the seroprevalence and risk factors associated with these three zoonoses among butchers and slaughterhouse workers in the Lorestan province of Iran. In 2017, 289 individuals (144 butchers or slaughterhouse workers, and 145 people from the general population) were enrolled in 11 different counties of this province. Collected serum samples were tested by ELISA for detection of IgG antibodies against Coxiella burnetii, Brucella spp. or Francisella tularensis antigens. The seroprevalence of Q fever, brucellosis and tularemia among all participants were 23.5%, 31.8% and 3.8%, respectively. The seroprevalence of brucellosis and Q fever among butchers and slaughterhouse workers (43.7% and 29.8%, respectively) were significantly higher (p < 0.05) than those of the general population (20% and 17.2%, respectively). A contact history with small ruminants (sheep and goats) was associated with a higher risk of positive serology for all three studied zoonoses. The high seroprevalence for Q fever and brucellosis we found among butchers and slaughterhouse workers suggests that both diseases are common in these populations of the Lorestan province. Since these two infectious diseases are clinically unspecific, they must be systematically included in the etiological diagnosis of infectious diseases occurring in these at-risk populations. In addition, we recommend specific training programs as well as the use of personal protective equipment in these occupational groups to reduce the occurrence of these zoonotic diseases.

2018 related Products with: Seroepidemiological study of Q fever, brucellosis and tularemia in butchers and slaughterhouses workers in Lorestan, western of Iran.

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Toxoplasma gondii in naturally infected goats: Monitoring of specific IgG levels in serum and milk during lactation and parasitic DNA detection in milk.

The zoonotic protozoa Toxoplasma gondii is one of the major abortive pathogens in small ruminants. Nevertheless, data on T. gondii infection in goats during lactation and on the presence of T. gondii in goat milk are lacking. A longitudinal study was planned in a T. gondii naturally infected dairy goat farm with the aim of (i) evaluating the variation of anti-T. gondii antibodies in blood and milk during the lactation; (ii) identifying the optimal phase during lactation for T. gondii monitoring; (iii) detecting the presence of T. gondii DNA in the milk. From March to July 2017, 30 goats in a farm were fortnightly visited seven times and sampled for blood and, when in lactation, for milk. Individual data regarding age, reproductive disorders, and the day of lactation were recorded. For the detection of anti-T. gondii antibodies in blood and milk a commercial ELISA kit was used. Milk samples (n = 63) of selected nine seropositive animals were also molecularly analysed to amplify a sequence within the ITS1 region of T. gondii. The seroprevalence of T. gondii infection was 63.3% (19/30); a high agreement was obtained between serum and milk results (Spearman's coefficient = 0.793 and Kendall's tau = 0.624), particularly between the 15 and the 60 day of lactation. In the statistical analysis, performed with generalized linear mixed models (GLMMs), the variable "phase of lactation" was strongly associated to ELISA values obtained in both serum and milk (p-value = 0.0001, F = 5.197, and p-value = 0.016, F = 2.755, respectively). Finally, molecular analyses revealed the presence of parasitic DNA in 20.6% (13/63) of milk samples, with a discontinuous parasite excretion; statistical analyses did not reveal any association among the parasite excretion and the considered variables. Milk could be considered as a valid alternative to blood for monitoring T. gondii infection in goat herds. Moreover, the detection of T. gondii DNA in milk enhanced the possibility for raw goat's milk consumption to be considered as a risk to public health.

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Immuno-analytical method development for detection of transgenic Cry1Ac protein and its validation.

Bacillus thuringiensis (Bt) synthesizes Cry1Ac protein, which is toxic to many lepidopteran pests, and the cry1ac gene has been expressed in several transgenic crop plants. The Cry1Ac protein has been isolated from Bt kurstaki HD73 and purified to homogeneity. Polyclonal antibodies were raised against purified Cry1Ac in rabbits and goat. Sandwich ELISA was developed for Cry1Ac using goat IgG as a coating antibody, and affinity-purified rabbit IgG as the primary antibody.

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Combined Use of Indirect ELISA and Western Blotting with Recombinant Hepatocellular Carcinoma-Associated Antigen 59 Is a Potential Immunodiagnostic Tool for the Detection of Prepatent Infection in Goat.

is recognized as one of the important health problems in small ruminants, leading to reduced production and economic loss for farmers worldwide. Prepatent diagnosis of infection is crucial to improve control strategies as this helminth may remove up to one-fifth of total erythrocytes and may cause anemia, edema, diarrhea, and ultimately death in young animals. In this study, one of the excretory and secretory products, rHc-HCA59, was purified and used as antigen to detect specific antibodies in infected goats during prepatent stage of infection using indirect enzyme linked immunosorbent assay (ELISA) as screening test. All goats ( = 38) were housed indoor, experimentally infected with 8000 infective larvae (L3) of , and serum samples were collected prior to infection and at 14th day of infection. Immunoblotting was performed to confirm the results of indirect ELISA, evaluate the cross reactivity against rHc-HCA59 in sera of most common co-infecting parasites and rectify the false negative samples. Furthermore, three different batches of rHc-HCA59 were produced to evaluate the repeatability of ELISA. No eggs were detected in feces of all goats collected at 7th and 14th day of infection but, eggs were detected at 21 days post infection in the feces. Indirect ELISA performed in this study showed 87% sensitivity and 100% specificity. The western blot analysis confirmed immunoreactivity in serum samples which scored positive in indirect ELISA and recognized the samples as negative which had OD lower than negative cut-off value in indirect ELISA. Furthermore, all false negative sera ( = 5) that had OD value between positive and negative cut-off value in rHc-HCA59 based ELISA were clearly positive in western blot. Moreover, no cross-reactivity was detected in ELISA and western blotting against rHc-HCA59 in positive sera of , and The results of this study concluded that combined use of indirect ELISA and western blotting with rHc-HCA59 is a potential immunodiagnostic tool for the detection of infection during prepatent period in goats.

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PTH immunoassay interference due to human anti-mouse antibodies in an obese subject with normal parathyroid function.

Immunoassay interference is most often found for Prolactin measurement. Only few data exist on immunoassay interference for other hormones.

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