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Background: The study aimed to assess the filter paper blood sampling technique for sero- monitoring against Hydro-pericardium syndrome (HPS). In view of the fact, dried blood samples don’t require immediate refrigeration, occupy little space and are easily transported. Methods: Whole blood paired with serum samples were collected from 100 broiler chickens on filter paper strips, dried for 2hrs at 37oC, stored in polythene bag and then eluated in normal saline at 4oC for overnight. Antigen of HPS was isolated, purified and confirmed by agar gel precipitation test (AGPT) with raised hyper-immune serum. Eluates of whole blood dried on filter paper with corresponding serum samples were tested for antibody activity by indirect heam-agglutination (IHA) test. Results: ‘The IHA titers of eluates were similar to titers obtained with serum diluted as 1:10. Normal saline and phosphate buffered saline did not influence the antibody stability, when used as eluating fluid. Whole blood dried on filter paper could be stored sealed in plastic bag at 4oC or ambient temperature for at least one week with no appreciable loss of antibody titers. A strong correlation (r = 0.900) exist between the titers obtained with two methods of blood sampling. Conclusion: This study demonstrated that recovery of antibodies from blood dried on filter paper after eluation produces results comparable to those obtained by recovering antibodies from serum. Based on above findings it is concluded that filter paper blood sampling could serve as a cost effective and convenient tool for HPS sero-monitoring.
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The Triângulo Mineiro region from Minas Gerais state, is an important meat-exporting region of Brazil and data about Toxoplasma gondii infection in pigs raised and slaughtered in this area are scarce. Therefore, the aim of this study was to evaluate the occurrence of T. gondii in swine and establish the risk factors associated with the infection. Samples were collected from 600 pigs raised under intensive system in farms located at three different counties (Carmo do Paranaíba, Patrocínio and Perdizes). The samples were submitted to indirect hemagglutination antibody test with dilution of 1:32 and to indirect immunofluorescence antibody test with a cutoff of 1:64. The occurrence of positive pig was 3.3% (n=20) and 51.8% (n=311) respectively. A significant difference was observed between toxoplasmatic infection and factors such as lineage, animal origin, size of the farm, collective raising with others species, presence of rodents and type of water offered (p≤0.05). There was no difference between gender and the farm goals. The results demonstrated an occurrence of anti-T.gondii antibodies higher than expected for intensive pig raising system on the studied area, which could indicate a possible sanitary management problem on the studied proprieties. Improvements on the raising techniques are necessary to reduce T. gondii infection sources.(AU)
A região do Triângulo Mineiro, no estado de Minas Gerais, é uma importante região exportadora de carne do Brasil e pesquisas sobre a infecção por Toxoplasma gondii em suínos criados e abatidos nesta região são escassos. Portanto, o objetivo deste estudo foi avaliar a ocorrência de T. gondii nesses animais e estabelecer os fatores de risco associados com a infecção. Foram coletadas amostras de 600 suínos criados sob sistema intensivo, em fazendas localizadas em três municípios diferentes (Carmo do Paranaíba, Patrocínio e Perdizes). As amostras foram submetidas à Hemaglutinação Indireta com diluição de 1:32 e à Reação de Imunofluorescência Indireta com ponto de corte 1:64. A ocorrência de suínos positivos foi de 3,3% (n=20) e 51,8% (n=311), respectivamente. Foi observada diferença significativa entre a infecção toxoplásmica e fatores como linhagem, procedência dos animais, tamanho das propriedades, criação em conjunto com outras espécies, presença de roedores e tipo de água consumida (p≤0,05). Não houve diferenças estatísticas entre o sexo e finalidade de produção em relação à infecção por T. gondii. Os resultados demonstraram uma ocorrência de anticorpos anti-T. gondii superior à esperada em criações intensivas de suínos na região estudada, o que poderia indicar uma possível falha no manejo sanitário das propriedades estudadas. Melhorias nas técnicas de criação são necessárias para redução das fontes de infecção por T. gondii nos rebanhos.(AU)
Subject(s)
Animals , Swine , Toxoplasma , Toxoplasmosis, Animal/etiology , Toxoplasmosis, Animal/epidemiology , Serology , Hemagglutination Tests/veterinary , Risk Factors , Fluorescent Antibody Technique, Indirect/veterinaryABSTRACT
Objective To analyze the antibody titer value of indirect haemagglutination test(IHA)in 135 confirmed acute schistosomiasis patients,so as to provide the evidence for improving the diagnosis and treatment of acute schistosomiasis. Meth-ods A total of 135 acute schistosomiasis inpatients were selected from 2001 to 2006. They all received the IHA antibody titer de-tection,and the correlation among the age,incubation period,and hospitalization days was calculated. Results The antibody titers of IHA were higher than 1:320 in all the cases. The percentages of 1:640,1:1280,1:2560,1:5120 and 1:10240 were 1.48%,28.15%,35.56%,20.00%,and 14.81%respectively. The mean age was(47.70 ± 14.58)years,average incuba-tion period was(38.03 ± 4.59)days and mean hospital stay time was(15.08 ± 3.79)days. The antibody titer value had no corre-lation with the age distribution(r=0.109,P>0.05). There was a negatively correlation between the antibody titer value and in-cubation period,(r=-0.558,P<0.01),there was a positive correlation between the antibody titer value and hospitalization time(r=0.791,P<0.01),and there were significant differences among different groups(F=17.07,64.53,both P<0.01). Conclusions The antibody titer of acute schistosomiasis cases detected by IHA is 1:640 and above. There is no correlation be-tween the antibody titer value and age,but the antibody titer value is higher,the incubation period is shorter and hospitalization time is longer.
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Haemorrhagic Septicaemia is one of the most common, fatal and acute bacterial diseases of livestock which causes mortality above 70% and is caused by Pasteurella multocida. The only satisfactory and practical method of control and prevention is timely vaccination of all the healthy and in contact animals. Different types of vaccines are being used for the immunity against this disease. In this project three oil based vaccines were produced. Two single emulsion vaccines were prepared by utilizing Montanide ISA-50 and liquid paraffin with lanolin where as one double emulsion with the help of Montanide ISA-206 was prepared. In house quality control testing and safety testing was performed on swiss albino mice. For immune titre IHA was performed by collecting serum from each and every animal including control animals. The comparison of IHA was done via statistical analysis by using GMT, Single emulsion vaccine prepared from liquid paraffin with lanolin gave maximum immune titre out of all the three vaccines in large animals and in young calves ISA 206 gave a significant titre.
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Objective To comprehensively evaluate the effects of indirect hemagglutination test(IHA),enzyme?linked im?munosorbent assay(ELISA),and dipstick dye method(DDIA)in the diagnosis of schistosomiasis japonica at different preva?lence by using Meta?analysis. Methods Through the literature review according to the inclusion and exclusion criteria,a data?base was established,and by using Meta?disc and R software,the Meta?analysis was performed including the threshold test,het?erogeneity test,weighted by the quantitative effect of merger,SROC curve fitting,etc. Results A total of 60 papers were in?cluded in the final analysis. The sensitivities of IHA were 0.84,0.76 and 0.94 in heavy,medium and low endemic areas,and specificities were 0.73,0.64 and 0.73 respectively;the sensitivities of ELISA were 0.88,0.80 and 0.93 in heavy,medium and low endemic areas,and the specificities were 0.59,0.59 and 0.62 respectively;the sensitivities of DDIA were 0.93,0.81 and 0.93 in the heavy,medium and low endemic areas,and specificities were 0.66,0.69 and 0.59 respectively. The weighted sensi?tivities of IHA,ELISA and DDIA were 0.83,0.87 and 0.90 respectively;the weighted specificities were 0.69,0.60 and 0.62 re?spectively. The areas under the curve of SROC were 0.89,0.96 and 0.92 in IHA,ELISA and DDIA respectively. Conclusions In different prevalence,the effectiveness of different methods for serological diagnosis of schistosomiasis is different. The sensi?tivity and specificity of all diagnostic methods of schistosomiasis need to further improve.
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Objective To understand the changes of positive rates of IHA detections of outpatients in schistosomiasis clinic. Results The data of IHA detections of outpatients in schistosomiasis clinic in Hubei Provincial Center for Disease Control and Prevention were collected and analyzed statistically from 2005 to 2014. Results A total of 7 113 outpatients were detected by IHA test,and 547 of them were positives with a positive rate of 7.69%. The positive rate of IHA test was on an upward slope be?fore 2008,and the rate reached 14.85% in 2008,which was significantly higher than that in 2005(5.81%)( χ2 = 47.40,P<0.01),then it was on a declined stage after 2008,and the positive rate decreased to 3.76 in 2014,which was significantly lower than that in 2008( χ2 = 12.29,P<0.01). The positive rate of outpatients in the 10~<30 years age group was higher than those in other age groups(all P < 0.012 5),and the male positives were more than the female ones. Conclusions The schisto?somiasis endemic situation has been significantly decreased in Hubei Province. The male and people in 10~<30 age group are the high risk groups,so the targeted health education should be strengthened.
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Objective To evaluate the value of indirect hemagglutination test(IHA)in schistosomiasis diagnosis. Meth?ods The literature concerned schistosomiasis diagnosis with IHA in the databases of Medline,CNKI,VIP and Wanfang Data from 1982 to 2014 was collected and evaluated. Results Totally 21 articles which were satisfied with the research criteria were analyzed with the Meta?analysis method. The IHA method had high value in schistosomiasis diagnosis,the AUCSROC of IHA in laboratory evaluation was 0.990 6,while in filed evaluation was 0.832 9,and the difference between them was significant(Z=4.50,P<0.05). Conclusion The diagnosis value of IHA in field evaluation is less than that in laboratory. In the process of the elimination of schistosomiasis,developing a new and higher sensitive reagent in schistosomiasis diagnosis is needed.
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Objective To explore the cut?off value of the indirect haemagglutination test(IHA)method for schistosomiasis japonica diagnosis in different endemic areas. Methods Totally 55 nature villages of the lake?type endemic counties,Yugan and Xinzi,in Poyang Lake Region of Jiangxi Province were chosen as the study fields,and all the villagers over 5 years old were parallelly examined by Kato?Katz method+miracidial hatching test and IHA method. The detection data were analyzed by the correlation analysis,and the threshold values of the IHA method in different endemic areas were decided by the receiver operat?ing characteristic(ROC)curve. Results The positive rate of stool examinations of the villagers was correlated with the distribu?tion trend of the antibody level of whole population(r=0.588,P0.05). The antibody level of stool?negative population during the period of 2008 to 2011 detect?ed by IHA method dropped year by year,and the annual difference was statistically significant(F=3.650,P0.05). When the positive rates were5%,the specificity of diagnosis could be improved when 1∶80,1∶20 and 1∶10 were used as the cut?off val? ues of IHA correspondingly. Conclusion The different threshold values for diagnosis of schistosomiasis japonica should be con?sidered while using IHA method to screen out patients in different endemic areas.
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Objective To establish written and electronic archives of Schistosoma japonicum antibody indirect hemagglutina-tion(IHA)tests. Methods In the process of schistosomiasis screening by IHA,the written records,electronic records,and se-rum sample bank were combined to make comprehensive archives. Results The S. japonicum antibody IHA test archives can pre-serve the schistosomiasis screening data in the long term and even can trace the source of experiments,and the operation was sim-ple. Conclusion The archives of S. japonicum antibody IHA tests are simple and useful,and worth of popularization.
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Objective To discuss the test efficiency of three methods for detecting Toxoplasma IgG antibody. Methods To-tally 304 specimens were detected parallelly for Toxoplasma IgG antibody by using the gold marked method,indirect hemagglutina-tion test(IHA),and enzyme-linked immunosorbent assay(ELISA),and the sensitivity,specificity and Youden index of these methods were compared. Results The detection sensitivities of gold marked method,IHA,and ELISA for Toxoplasma IgG anti-body were 85.5%,89.8%and 91.9%respectively(χ2=4.12,P>0.05);the specificities were 92.4%,96.6%and 97.5%respec-tively(χ2=4.06,P>0.05). The detection efficiency and Youden index of ELISA were 94.1%and 0.89 respectively,being high-er than those of IHA and gold marked method. Conclusion The sensitivity and specificity of the ELISA method for Toxoplasma IgG antibody are higher,and in addition,it can be automated. Therefore,it is suitable for large-scale Toxoplasma IgG antibody screening.
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Objectives To explore the correlation between the population characteristics of sero-reactivity with quantitative antibody based-IHA and the transmission parameters,such as epidemic situation,transmission status or infection trend in population.Methods The residents in one endemic administrative village were simultaneously examined by Kato-Katz technique for parasitological stool examination,as well as by immuno-diagnostic technique IHA for detection of IgG antibody against soluble egg antigen for two consecutive years.The results of examination were analyzed and compared on the diagnostic parameters of IHA,the correlation of the changes of positive rates and antibody levels of IHA with the changes of infection trend in population and the distribution of antibody levels between 'the true negative' and 'the true positive'.Results When Kato-Katz technique based on 2 stool samples,each read in 3 thick smears,was used as the reference,the overall sensitivity of IHA was high (from 77.27% to 85.48%) with a relatively poor specificity of lower than 60%,the negative predict value (NPV) was excellent of higher than 94%.The specificity of IHA decreased with the increase of the age in different age-groups of population,showing the highest among the younger less than 15 years old.The distribution trends of positive rates of antibody in different age groups by IHA showed similar to that of egg positive rate detected by Kato-Katz although the positive rates of IHA were higher than these by Kato-Katz,which showed that a higher false positive (from 41.90% to 44.56%) and a certain false negative (from 14.52% to 22.73%)existed in IHA.The positive rate of antibody decreased slowly among the individuals with S.japonicum infection,who received treatment.There was an overlap in the distribution of antibody levels between ' the true negative' and ' the true positive'.Conclusions Under the current schistosomiasis epidemic situation in China,IHA is valuable in the epidemiologic surveys.It should be of further deliberation applying IHA as the screening approaches for identification of target individuals for treatment or determination of the infection rate in community and IHA needs to be combined with the parasitological examination.
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To establish a definite diagnosis for pulmonary hydatid disease, combination of radiology and serology is useful. In this study, 19 preoperative sera from patients with surgically confirmed pulmonary hydatidosis, 40 sera from patients with other parasitosis and pulmonary diseases, and 20 sera from healthy donors were evaluated using 4 different serological tests, i.e., the commercial ELISA (ELISA-kit) test, the ELISA (ELISA-lab) test prepared in our laboratory, the commercial indirect hemagglutination assay kit (IHA-kit) test, and the IHA test using sensitized sheep red blood cells with tannic acid (IHA-TA). The ELISA-kit was the most sensitive (84.2%) and the most specific test (100.0%). The ELISA-kit also demonstrated the highest positive (100.0%) and negative (95.2%) predictive values. The sensitivity of the ELISA-lab test, that we prepared, was found to be 73.6%, whereas the IHA-kit test and the IHA-TA test were found to be 73.6% and 68.4%, respectively. The specificity of these tests was 96.6%, 98.3%, and 83.3%, respectively. When all 4 tests were assessed together, it was found that the sensitivity had risen to 94.7%. When the ELISA-kit was assessed with the IHA-kit and IHA-TA together, it was found that the sensitivity was 89.5% and 84.2%, respectively. Likewise, the combination of the ELISA-lab and IHA-kit or IHA-TA allowed us to achieve a sensitivity of 84.2% in cases of pulmonary echinococcosis. In conclusion, the diagnosis would be imminent if least 2 tests were applied together.
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Humans , Echinococcosis, Pulmonary/diagnosis , Enzyme-Linked Immunosorbent Assay/methods , Hemagglutination Tests/methods , Predictive Value of Tests , Sensitivity and Specificity , Serologic Tests/methodsABSTRACT
Objective To investigate the family clustering of Toxoplasma infections. Methods The enzyme-linked immunosorbent assay (ELISA) and indirect hemagglutination test (IHA) were performed to detect antibodies to To.xoplasma in sera of 1 436 people from 487 families with clinical cases. The data were analysed with G test of homogeneous in statistical binomial distribution. Results Three hundred and forty-two people of 237 families showed sera positive, with rates of 48.7% of family and 23.8% of population. A saliva trophozoite in one of 30 rats was found and so were two typical saliva parasites in one of 10 cases, Conclusion There is a significant evidence of family clustering of Toxoplasma infections (T=6.48,P