Seroprevalence of Anti-Mers-CoV IgG in two groups of Qatar population: Blood donors and case contacts

Since its first isolation in September 2012, Middle East respiratory syndrome coronavirus (MERS-CoV) has diffused across 27 countries infecting more than 1910 individuals with a high case fatality rate. However, MERS-CoV has also been reported to be asymptomatic or to cause influenza-like mild illnesses. In the absence of clear epidemiological view, cross-sectional MERS-CoV antibody surveillances in human populations are of global concern. In this study, we present a comparative serological screening of 4719 blood donors, 135 baseline case contacts and 4 MERS-CoV confirmed patients for the presence of anti-MERS-CoV IgG. Methods: Samples were initially screened using MERS-CoV recombinant spike protein Enzyme linked immunoassay (rELISA) from Euroimmune, Germany. To confirm rELISA results, farther serological testing has been performed for borderline and reactive anti-MERS-CoV IgG samples by indirect immunofluorescent test (full virus IIFT) IgG/M, recombinant spike protein indirect immunofluorescent assay IgG (rIIFA) and pseudovirus neutralizing assay (ppNT). To access cross reactivity, borderline and reactive samples were also tested for presence of IgG to other human coronaviruses (HCoV) using IIFT, rIIFA and/ or in house rELISA. Results: rELISA yielded 3 borderlines (all donors) and 12 reactive (7 donors, 1 case contact and 4 samples collected from 3 MERS-CoV confirmed patients) anti-MERS-CoV IgG results. However, IIFT IgG confirmed only 5 reactive rELISA results (2 blood donors and 3 patients; the reactive case contact was not sufficient to be tested by IIFT IgG). Yet, r-IIFA and ppNT only confirmed the presence of specific anti-MERS-CoV antibodies in patients’ samples. Interestingly, all borderline and reactive tested samples showed reactive titers against recombinant spike proteins of other HCoV. Conclusion: Our findings suggest that MERS-CoV is not heavily circulated among the population of Qatar. This study provides an insight about the epidemiological view for MERS-CoV in Qatar population. It also provides a performance evaluation for the available serologic tests for MERS-CoV in a view of serologic status to other human coronaviruses

Comparative serology techniques for the diagnosis of Trypanosoma cruzi infection in a rural population from the state of Querétaro, Mexico

Immunological diagnostic methods for Trypanosoma cruzi depend specifically on the presence of antibodies and parasitological methods lack sensitivity during the chronic and “indeterminate” stages of the disease. This study performed a serological survey of 1,033 subjects from 52 rural communities in 12 of the 18 municipalities in the state of Querétaro, Mexico. We detected anti-T. cruzi antibodies using the following tests: indirect haemagglutination assay (IHA), indirect immunofluorescence assay (IFA), ELISA and recombinant ELISA (rELISA). We also performed Western blot (WB) analysis using iron superoxide dismutase (FeSOD), a detoxifying enzyme excreted by the parasite, as the antigen. Positive test results were distributed as follows: ELISA 8%, rELISA 6.2%, IFA and IHA 5.4% in both cases and FeSOD 8%. A comparative study of the five tests was undertaken. Sensitivity levels, specificity, positive and negative predictive values, concordance percentage and kappa index were considered. Living with animals, trips to other communities, gender, age, type of housing and symptomatology at the time of the survey were statistically analysed using SPSS software v.11.5. Detection of the FeSOD enzyme that was secreted by the parasite and used as an antigenic fraction in WBs showed a 100% correlation with traditional ELISA tests

Prevalence and molecular identification of Chlamydia abortus in commercial dairy goat farms in a hot region in Mexico

The aim of this study was to determine the seroprevalence and presence of Chlamydia abortus in Saanen breed female goats from commercial dairy goat farms under intensive production in the municipality of Guanajuato, Mexico. Sera were collected to determine the prevalence of anti-C. abortus IgG antibodies using recombinant enzymelinked immunosorbent assay (rELISA) and cell culture. Polymerase chain reaction (PCR) was used to prove the presence of the pathogen in swab samples collected from the vagina and rectum of selected animals. Additionally, foetal tissue samples from a sudden abortion were collected. C. abortus prevalence in female goats of commercial milking farms sampled in Guanajuato, Mexico, was 4.87 % (n=246). Seropositive animals were found in six out of nine (66.6 %) dairy goat farms sampled, and prevalence among animals in individual farms ranged between 3.44 and 13.51 %. C. abortus was detected using PCR in spleen tissue from the aborted foetus. PCR-based detection, as well as isolation from vaginal and rectal swabs, was not possible in the present study. Isolation through cell culture was also unsuccessful from aborted foetal tissue samples. In conclusion, the results from rELISA and PCR show that C. abortus is present in dairy goat farms in the state of Guanajuato, Mexico

Allergen-Specific Immunotherapy Follow-Up by Measuring Allergen-Specific IgG as an Objective Parameter

The clinical efficacy of the allergen‐specific immunotherapy (AIT) has been well‐documented using inhalant or hymenoptera‐derived allergens in atopic patients with corresponding specific IgE antibodies. AIT is considered as the unique treatment that is capable of modifying the natural course of the allergic disease because it induces a variety of immunological mechanisms, with emphasis in the production of blocking IgG antibodies by IL‐10‐stimulated B cells due to the generation of Treg, Breg, or even Th2 cells. Thus, the measurement of specific IgG subclasses, particularly IgG4, to the crude extract or more importantly to allergen components, might be a useful and potential tool to follow‐up objectively the patients undergoing AIT in addition to clinical parameters. In this chapter, the authors have emphasized a very sensitive and highly specific reverse ELISA, developed by them, to measure IgG subclasses directed to clinically relevant natural allergens that are undoubtedly better when compared to those obtained with recombinant counterparts. Such a technique may produce more authentic results taking into account the IgG subclass binding capacity to a particular allergen and might be a valuable and alternative method for monitoring activation of tolerance‐inducing mechanisms in patients under AIT

Feline hemoplasmas : evaluation of specific antibodies and the molecular and cytological diagnostic

Dissertação de Mestrado Integrado em Medicina VeterináriaMycoplasma haemofelis, Candidatus Mycoplasma haemominutum and Candidatus Mycoplasma turicensis are three hemoplasmas responsible for feline hemolytic anemias. The diagnosis of the infection can be performed by observation of the agents on blood smears or by their identification and quantification using PCR techniques, presently considered the gold standard method. A recombinant antigen for Mhf (rDnaK) has been identified, characterized, produced and then successfully applied in Western blot and ELISA assays (rELISA) to detect antibodies in samples from experimentally induced infections. To evaluate this application under field conditions, this rELISA was used in samples collected from naturally infected cats, in parallel with quantitative PCR and cytologic examination. Within the scope of the TNR program implemented by the Lisbon City Council, blood samples were collected from 104 cats. After sample collection, blood was immediately used to prepare blood smears which were then stained with Giemsa (Parasitology Lab, FMV/Lisbon University). The remaining blood was split into two aliquots: 1 – Plasma separation, shipped to Vetsuisse Faculty, Zurich University, for antibody testing with rELISA; 2 – Total DNA extraction for identification and quantification of the three species of hemoplasma using qPCR (Virology Lab, FMV/Lisbon University). Out of 22.1% (N=23) samples where the microorganism was identified by qPCR, mycoplasmas were also identified on the blood smears of 15.4% (N=16). 4.8% (N=5) of samples tested as seropositive and 2.9% (N=3) revealed bordeline results. From 77.9% (N=81) of samples that were negative by qPCR, only 44.2% (N=46) were also negative on cytologic examination. 60.6% (N=63) of samples were considered seronegative and 6.7% (N=7) were borderline. Sensitivity for cytology was 69.6% and specificity was 56.8%. Sensibility of rELISA was 25% and its specificity was 85.1%. Cohen’s Kappa (k) was calculated to assess agreement between PCR-Cytology (k=0.1836) and PCR-rELISA (k=0.1093). Given the low agreement, PCR was found to be the most appropriate diagnostic method. Further studies are necessary to characterize the response of the immune system and the role of different antigens in these infections in order to improve the suitability of rELISA in a clinical settingRESUMO - HEMOPLASMAS FELINOS: AVALIAÇÃO DE ANTICORPOS ESPECÍFICOS E CORRESPONDÊNCIA COM O DIAGNÓSTICO MOLECULAR E CITOLÓGICO - Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum e Candidatus Mycoplasma turicensis são três hemoplasmas responsáveis por anemias hemolíticas em felinos. O diagnóstico desta infecção pode ser feito pela observação dos agentes em esfregaços de sangue ou através da sua identificação e quantificação pelas técnicas de PCR, sendo este ultimo considerado o método gold standard. Um antigénio recombinante do Mhf (rDnaK) foi identificado, caracterizado, produzido e posteriormente aplicado com sucesso, em técnicas de Western blot e rELISA, para a deteção de anticorpos em amostras colhidas de gatos infetados experimentalmente. De forma a avaliar a sua aplicação numa realidade clínica, esta rELISA foi utilizada em amostras colhidas de gatos naturalmente infectados, em paralelo com o PCR quantitativo e com o exame citológico. No âmbito do programa CED efetuado pela Câmara Municipal de Lisboa, foi realizada a recolha de sangue a 104 gatos. Após a colheita das amostras, o sangue foi utilizado para preparar esfregaços, corados de seguida com coloração Giemsa (Laboratório de Parasitologia, FMV/ULisboa), e dividido em duas alíquotas: 1- Separação do plasma, que foi enviado para a Faculdade Vetsuisse, Universidade de Zurique, onde realizaram a pesquisa de anticorpos com a rELISA; 2 – Extracção do ADN total para a identificação e quantificação das três espécies de hemoplasmas por qPCR (Laboratório de Virologia, FMV/ULisboa). Das 23 (22.1%) amostras PCR positivas, foram igualmente encontrados micoplasmas em 16 esfregaços (15.4%). 5 (4.8%) amostras foram consideradas seropositivas e 3 (2.9%) apresentaram resultados inconclusivos. Das 81 (77.9%) amostras negativas ao PCR, apenas 46 (44.2%) mostraram-se negativas ao exame citológico. 63 (60.6%) amostras foram consideradas seronegativas e 7 (6.7%) apresentaram valores não conclusivos. A sensibilidade da citologia foi de 69.6% e a especificidade foi de 56.8%. Por sua vez, a rELISA apresentou uma sensibilidade de 25% e uma especificidade de 85.1%. O Cohen’s Kappa (k) foi calculado para avaliar a concordância entre o PCR e a Citologia (k=0.1836) e o PCR e a rELISA (k=0.1093). Tendo em conta as baixas concordâncias, o nosso estudo confirma que o PCR é o método de diagnóstico mais adequado. Mais estudos são necessários para caracterizar a resposta imunitária e o papel dos vários antigénios destes agentes, de forma a melhorar a sua aplicação no rELISA e esta poder ser usada no contexto clínico.N/

Chagas Disease among the Latin American Adult population attending in a primary care center in Barcelona, Spain

Background/Aims: The epidemiology of Chagas disease, until recently confined to areas of continental Latin America, has undergone considerable changes in recent decades due to migration to other parts of the world, including Spain. We studied the prevalence of Chagas disease in Latin American patients treated at a health center in Barcelona and evaluated its clinical phase. We make some recommendations for screening for the disease. Methodology/Principal Findings: We performed an observational, cross-sectional prevalence study by means of an immunochromatographic test screening of all continental Latin American patients over the age of 14 years visiting the health centre from October 2007 to October 2009. The diagnosis was confirmed by serological methods: conventional in-house ELISA (cELISA), a commercial kit (rELISA) and ELISA using T cruzi lysate (Ortho-Clinical Diagnostics) (oELISA). Of 766 patients studied, 22 were diagnosed with T. cruzi infection, showing a prevalence of 2.87% (95% CI, 1.6-4.12%). Of the infected patients, 45.45% men and 54.55% women, 21 were from Bolivia, showing a prevalence in the Bolivian subgroup (n = 127) of 16.53% (95% CI, 9.6-23.39%). All the infected patients were in a chronic phase of Chagas disease: 81% with the indeterminate form, 9.5% with the cardiac form and 9.5% with the cardiodigestive form. All patients infected with T. cruzi had heard of Chagas disease in their country of origin, 82% knew someone affected, and 77% had a significant history of living in adobe houses in rural areas. Conclusions: We found a high prevalence of T. cruzi infection in immigrants from Bolivia. Detection of T. cruzi¿infected persons by screening programs in non-endemic countries would control non-vectorial transmission and would benefit the persons affected, public health and national health systems

Kajian pendanaan pendidikan pusat dan daerah

Dalam rangka memenuhi kebutuhan sarana prasarana pendidikan yang memadai, Pemerintah telah menyalurkan dana baik yang bersumber dari Pemerintah Pusat maupun Pemerintah Daerah. Penyaluran dana dari Pemerintah Pusat dilakukan melalui alokasi langsung ke sekolah berupa Bantuan Sosial/Bantuan Pemerintah (Bansos/Bantah), sedangkan penyaluran dana melalui transfer daerah adalah dalam bentuk Dana Alokasi Khusus (DAK). Sejauh ini pemanfaatan DAK dinilai masih belum optimal terbukti belum terserapnya sebagian besar DAK yang disalurkan, sementara itu masih banyak sekolah yang belum memiliki sarana prasarana yang memadai untuk penyelenggaraan proses pembelajaran. Selain itu, penyaluran dana berupa Bansos/Bantah masih belum sepenuhnya tepat sasaran. Oleh karena itu, pada tahun 2017 Pusat Penelitian Kebijakan Pendidikan dan Kebudayaan, Balitbang melaksanakan Kajian Pendanaan Pendidikan Pusat dan Daerah, khususnya dalam hal penyaluran Bansos/Bantah dan DAK pada jenjang pendidikan dasar dan menengah, yaitu SD, SMP, SMA, dan SMK. Tujuan dilakukannya kajian ini adalah untuk memperoleh gambaran tentang perkembangan alokasi dana Bansos/Bantah dan DAK, pemanfaatannya di sekolah, kendala yang dihadapi serta upaya yang dilakukan sekolah untuk mengatasi kendala yang ada. Berdasarkan hasil kajian dikembangkan opsi kebijakan untuk memperbaiki mekanisme penyaluran Bansos/Bantah dan DAK yang lebih efektif dan tepat sasaran

PSIKOEDUKASI KETERAMPILAN REGULASI EMOSI PADA SISWA SMP

Pada masa perkembangan remaja individu memasuki masa storm and stress. Kemampuan regulasi emosi yang berlangsung tidak tepat, akan membuat remaja cenderung mengikuti emosinya dalam berbagai tindakan yang menyebabkan hal-hal yang tidak diinginkan terjadi, seperti terjadinya perkelahian yang merugikan lingkungan sekitar. Keterampilan psikologis akan membantu siswa untuk meningkatkan kesehatan mental mereka. Pengabdian kepada masyarakat yang berupa psikoedukasi dilakukan pada siswa Sekolah Menengah Pertama. Sebanyak 47 siswa mengikuti kegiatan ini. Hal ini ditunjang dari asesmen kebutuhan melalui wawancara dan menyebarkan kuesioner berupa google form dilakukan untuk mengidentifikasi masalah yang di alami siswa. Psikoedukasi memberikan pengetahuan dan keterampilan (role play) metode regulasi emosi dengan Expressive Writing dan Breathing Exercise. Berdasarkan analisis yang dilakukan dengan menggunakan uji paired sample t-test menunjukkan peningkatan dari nilai pre-test sebesar 33.000 menjadi 33.979 dalam perhitungan post-test pemahaman tentang psikoedukasi regulasi emosi. Sedangkan dalam pre-test Visual Scale Analoge (VAS) untuk mengukur emosi yang semulanya memiliki rata-rata 3.340 memiliki penurunan menjadi 3.149. Hasil ini menggambarkan bahwa psikoedukasi keterampilan regulasi emosi mampu menyelesaikan persoalan-persoalan yang dihadapkan siswa SMP terutama untuk meningkatkan keterampilan meregulasi emos