Search results for: ELISA test

Authors: Zakieh Rostamzadeh , Nariman Sepehrvand-Zahra Shirmohamadi

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Background: Cytomegalovirus (CMV) infection is one of the most common infectious problems following kidney transplantation. In this study, we are aimed to investigate the CMV infection in the setting of renal transplant recipients in Urmia-Iran, using both ELISA and polymerase chain reaction (PCR) methods. Methods: Ninety-six renal transplant recipients were selected randomly and enrolled in a cross-sectional study. Blood sampling was done via venipuncture, and all sera were investigated for anti-CMV IgM, and the seropositive cases in association with 14 randomly selected seronegative cases were investigated with PCR assay. Results: Thirty-three patients (34.3%) were seropositive for anti-CMV IgM, 3 patients (3.1%) were in borderline range, and 60 patients (62.5%) were seronegative. By considering the patients with borderline anti-CMV IgM levels as seropositive, 37.5% were seropositive for anti-CMV IgM. Among 36 seropositive cases, the CMV infection was confirmed in 19 (52.7%) of them using PCR. Age (P = 0.40), educational status (P = 0.77), history of pre-transplantation dialysis (0.52), history of blood transfusion (P = 0.52), and immunosuppressive regimen were not statistically different among recipients with positive versus negative CMV PCR study results. Conclusion: The seroprevalence of CMV infection was demonstrated to be high in renal transplant recipients of Urmia-Iran. The rate was higher compared to several previous reports in the literature. ELISA method has an appropriate sensitivity to screen the recipients for CMV infection but considering its relatively low specificity, the seropositive cases are better to be confirmed by further PCR study.

Keywords: cytomegalovirus, renal transplantation, ELISA, IgM, PCR

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Authors: Gulnara Azizova, Naila Hasanova, Nazenin Hasanzade

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The aim of study is to investigate the role of osteoactivin as a more sensitive and modern diagnostic biomarker that has a prognostic value in metabolic and repair processes occurring in bone and cartilage tissue in osteoporosis and osteoporotic fractures. Osteoactivin (OA) is a new glycoprotein that is highly expressed during osteoblast differentiation. It was first discovered in an osteopetrotic rat model using mRNA . This study was carried out on patients between the ages of 45-83 from the Department of Traumatology and placed in 3 groups: group I - 14 patients with osteoporosis, group II - 15 patients with non-osteoporotic fractures, group III - 25 patients with osteoporotic fractures. The control group consisted of 14 healthy people. To monitor changes in osteoactivin, blood samples were taken at 3 stages: on day 1 before treatment, on day 10 of treatment, and 1 month after treatment. The concentration of OA in the blood serum was determined by ELISA method on the immunoassay analyzer “Mindray MR- 96A” using a set of reagents from the company Boster ( ELISA Kit PicoKine, USA). The statistical evaluation was performed by using SPSS 22.0 program (IBM SPSS Inc., USA). Compared to the control, osteoactivin concentration increased by 66.2% in patients with osteoporosis, 54.1% in patients with non-osteoporotic fractures, and 80.2% in patients with osteoporotic fractures, indicating that it plays an important role in the pathogenesis of osteoporotic fractures. At 1 month after treatment, osteoactivin concentration increased by 81.6% in patients with non-osteoporotic fractures. The lack of a significant change in osteoporotic fractures is explained by the late healing of these fractures. Based on the sensitivity and specificity indicators, the ROC curve was created and it was determined that osteoactivin is a test with high general diagnostic value, specificity and informativeness in the prognosis of osteoporosis and osteoporotic fractures, and can be used throughout the treatment period.

Keywords: osteoactivin, bone, osteoporosis., cartilage

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Authors: Ibrahim Djelloul Berkane

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A survey was conducted to assess the presence of the virus in Citrus tristeza one of the main citrus regions of Algeria, namely the Chlef region, using the technique of Direct Tissue Print Immunoprinting Assay (DTBIA) and the Double Sandwich ELISA antibodies. A nursery citrus, lumber yards, and commercial orchards, which are the main varieties cultivated citrus were subjected to samples collected samples for laboratory analysis. 0.91% of the plants tested orchards were infected with CTV, while no positive case was detected at the nursery the yard, however, it is reported that an alarming rate of 10,5% of orchards tested at the common Chettia were infected with tristeza virus. The investigation was launched to identify the vector species tristeza revealed the presence of a vector is important Aphis gossypii.

Keywords: aphis, chlef, citrus, DAS-ELISA, DTBIA, tristeza

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Authors: Hiroshi Nakayama, Yuji Ito

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Synthetic cannabinoids have attracted much public attention recently in Japan. 1-pentyl-3-(1-naphthoyl)-indole (JWH-018), 1-pentyl-2-methyl-3-(1-naphthoyl) indole (JWH-015), 1-(5-fluoropentyl)-3- (1-(2,2,3,3- tetramethylcyclopropyl)) indole (XLR-11) and 1-methyl-3- (1-admantyl) indole (JWH-018 adamantyl analog) are known as synthetic cannabinoids and are also considered dangerous illegal drugs in Japan. It has become necessary to develop sensitive and useful methods for detection of synthetic cannabinoids. We produced two monoclonal antibodies (MAb) against synthetic cannabinoids, named NT1 (IgG1) and NT2 (IgG1), using Hybridoma technology. The cross-reactivity of these produced MAbs was evaluated using a competitive enzyme-linked immunosorbent assay (ELISA). In the results, we found both of these antibodies recognize many kinds of synthetic cannabinoids analog. However, neither of these antibodies recognizes naphtoic acid, 1-methyl-indole and indole known as a raw material of synthetic cannabinoid. Thus, the MAbs produced in this study could be a useful tool for the detection of synthetic cannabinoids.

Keywords: ELISA, monoclonal antibody, sensor, synthetic cannabinoid

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Authors: Youn-Sung Kim, Jin-Ho Jo, Mi-Sung Kim, Jae-Kun Lee

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Accelerated life cycle test is applied to various products or components in order to reduce the time of life cycle test in industry. It must be considered for many test conditions according to the product characteristics for the test and the selection of acceleration parameter is especially very important. We have carried out the general life cycle test and the accelerated life cycle test by applying the acceleration factor (AF) considering the characteristics of brushless DC (BLDC) motor for washing machine. The final purpose of this study is to verify the validity by analyzing the results of the general life cycle test and the accelerated life cycle test. It will make it possible to reduce the life test time through the reasonable accelerated life cycle test.

Keywords: accelerated life cycle test, reliability test, motor for washing machine, brushless dc motor test

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Authors: Hassan Waseem

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Cytomegalovirus (CMV) is ubiquitously distributed viral agent responsible for different clinical manifestations that may vary according to the immunologic status of the patient. CMV can cause morbidity and mortality among fetuses and patients with compromised immune system. A cross-sectional study was carried out in Islamabad to investigate the prevalence and risk factors associated with CMV infection among pregnant women. Blood samples of 172 pregnant women visiting Mother and Child Healthcare, Pakistan Institute of Medical Sciences (PIMS) Islamabad were taken. In present study, serum samples of the women were checked for CMV-specific IgG and IgM antibodies by enzyme linked immunosorbent assay (ELISA). Clinical, obstetrical and socio-demographical characteristics of the women were collected by using structured questionnaires. Out of 172 pregnant women included in the study, 171 (99.4%) were CMV specific IgG positive and 30 (17.4%) were found positive for CMV-IgM antibodies. The CMV has taken an endemic form in Pakistan so, routine screening of CMV among pregnant women is recommended.

Keywords: Cytomegalovirus, blood transfusion, ELISA, seroprevalence

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Authors: Xiaohuan Li, Wangmin Yi, Xinghui Wu

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In order to verify the performance of lunar lander structure, landing-impact test is urgently needed. Moreover, the test equipment is necessary for the test. The functions and the key points of the equipment is presented to satisfy the requirements of the test，and the design scheme is proposed. The composition, the major function and the critical parts’ design of the equipment are introduced. By the load test of releasing device and single-beam hoist, and the compatibility test of landing-impact testing system, the rationality and reliability of the equipment is proved.

Keywords: landing-impact test, lunar lander, releasing device, test equipment

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Authors: Youjeong Hwang

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Purpose: Insulin-like growth factor-I (IGF-I) promotes B-cell development, immunoglobulin formation, and interleukin-6 (IL-6) production, then regulate the immune response and inflammation. As IGF-I and their receptor also exist in the periodontal tissue, they may affect the immune response caused by periodontal pathogens in aggressive periodontitis (AgP) patients. The function of IGF is regulated by IGF binding proteins (IGFBPs), and IGFBP-3 is known to most abundant in plasma. The aim of the present study was to assess the concentration of IGF-I and IGFBP-3 in plasma and gingival crevicular fluid (GCF) in AgP patients and to find out their association. Methods: Nine patients with AgP (test group) and nine healthy subjects (control group) were included in this study. None of the subjects had a history of systemic disease, smoking or steroids medication. GCF samples were collected by microcapillary pipettes and plasma samples were obtained by venipuncture. Probing pocket depth (PD), clinical attachment level (CAL) and bleeding on probing (BOP) were recorded. Samples were assayed for IGF-I and IGFBP-3 levels using ELISA. Results: Mean IGF-I level in GCF was higher in the test group than control. Mean IGF-I level in plasma and IGFBP-3 level in GCF and plasma in control group were higher than that of the test group. However, there was no statistical significance (p > 0.05). The mean level of IGF-I and IGFBP-3 in GCF was lower than those in plasma. Mean IGF-I level in plasma showed a negative correlation with PD and CAL (p < 0.05) in both groups. The levels of IGF-I and IGFBP-3 in GCF seemed to be negatively correlated with BOP in the test group (p < 0.05). Conclusions: The difference in the level of IGF-I and IGFBP-3 between AgP and healthy subjects was not significant. Further studies that explain the mechanism of the protective role of IGF-I with more samples are needed.

Keywords: aggressive periodontitis, pathogenesis, insulin-like growth factor, insulin-like growth factor binding protein

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Authors: Andrei Contan, Richard Torkar

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Aim: The process of test automation and its practices in industry have to be better understood, both for the industry itself and for the research community. Method: We conducted a quantitative industry survey by asking IT professionals to answer questions related to the area of test automation. Results: Test automation needs and practices vary greatly between organizations at different stages of the software development life cycle. Conclusions: Most of the findings are general test automation challenges and are specific to small- to medium-sized companies, developing software applications in the web, desktop or mobile domain.

Keywords: survey, testing, test automation, status of test automation

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Authors: Ghebremedhin Tsegay, Weldu Tesfagaber, Yuanmao Zhu, Xijun He, Wan Wang, Zhenjiang Zhang, Encheng Sun, Jinya Zhang, Yuntao Guan, Fang Li, Renqiang Liu, Zhigao Bu, Dongming Zhao*

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African swine fever (ASF) is a highly infectious viral disease of pigs, resulting in significant economic loss worldwide. As there is no approved vaccines and treatments, the control of ASF entirely depends on early diagnosis and culling of infected pigs. Thus, highly specific and sensitive diagnostic assays are required for accurate and early diagnosis of ASF virus (ASFV). Currently, only a few recombinant proteins have been tested and validated for use as reagents in ASF diagnostic assays. The most promising ones for ASFV antibody detection were p72, p30, p54, and pp62. So far, three ELISA kits based on these recombinant proteins have been commercialized. Due to the complex nature of the virus and variety forms of the disease, robust serodiagnostic assays are still required. ASFV p22 protein, encoded by KP177R gene, is located in the inner membrane of viral particle and appeared transiently in the plasma membrane early after virus infection. The p22 protein interacts with numerous cellular proteins, involved in processes of phagocytosis and endocytosis through different cellular pathways. However, p22 does not seem to be involved in virus replication or swine pathogenicity. In this study, E.coli expressed recombinant p22 protein was used to generate a monoclonal antibody (mAb), and its potential use for the development of blocking ELISA (bELISA) was evaluated. A total of 806 pig serum samples were tested to evaluate the bELISA. Acording the ROC (Reciever operating chracteristic) analysis, 100% sensitivity and 98.10% of specificity was recorded when the PI cut-off value was set at 47%. The novel assay was able to detect the antibodies as early as 9 days post infection. Finaly, a highly sensitive, specific and rapid novel p22-mAb based bELISA assay was developed, and optimized for detection of antibodies against genotype I and II ASFVs. It is a promising candidate for an early and acurate detection of the antibodies and is highly expected to have a valuable role in the containment and prevention of ASF.

Keywords: ASFV, blocking ELISA, diagnosis, monoclonal antibodies, sensitivity, specificity

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Authors: Ghada A. Gamea, Nabila A. Yaseen, Ahmed A. Othman, Ahmed S. Tawfik

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Infection with Toxoplasma gondii can cause serious complications in pregnant women, leading to abortion, stillbirth, and congenital anomalies in the fetus. Definitive diagnosis of T. gondii acute infection is therefore critical for the clinical management of a mother and her fetus. This study was conducted on 250 pregnant females in the first trimester who were inpatients or outpatients at Obstetrics and Gynaecology Department at Tanta University Hospital. Screening of the selected females was done for the detection of immunoglobulin (IgG and IgM), and all subjects were submitted to history taking through a questionnaire including personal data, risk factors for Toxoplasma, complaint and history of the present illness. Thirty-eight samples, including 18 IgM +ve and 20 IgM-ve cases were further investigated by the avidity and IgE ELISA tests. The seroprevalence of toxoplasmosis in pregnant women was (42.8%) based on the presence of IgG antibodies in their sera. Contact with cats and consumption of raw or undercooked meat are important risk factors that were associated with toxoplasmosis in pregnant women. By serology, it could be observed that in the IgM +ve group, only one case (5.6%) showed an acute pattern by using the avidity test, though 10 (55.6%) cases were found to be acute by the IgE assay. On the other hand, in the IgM –ve group, 3 (15%) showed low avidity, but none of them was positive by using the IgE assay. In conclusion, there is no single serological test that can be used to confirm whether T. gondii infection is recent or was acquired in the distant past. A panel of tests for detection of toxoplasmosis will certainly have higher discriminatory power than any test alone.

Keywords: diagnosis, serology, seroprevalence, toxoplasmosis

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Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

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In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

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Authors: Ibrahim Aly, Waleed Elawamy, Hanan Taher, Amira Matar

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Schistosomiasis is infection with blood flukes of the genus Schistosoma, which are acquired trans-cutaneously by swimming or wading in contaminated freshwater. The present study was proposed to produce ultra-sensitive, field-friendly high-throughput rapid immunochromatography diagnostic device for accurate detection of asymptomatic parasite carriers in schistosomiasis pre-elimination settings.For assessing diagnostic potential of rapid device, 50 blood samples from patients with schistosomiasis mansoni, 29 other proven parasitic diseases and 25 blood samples as negative control were from healthy individuals were used. The sensitivity of Quantitative antigen-capture nano-ELISAwas 82 %, and specificity was 87.1 %, where the sensitivity of Nano Dot- ELISA was 86 % and specificity was 90.7 %. The sensitivity of diagnostic device was 78 % and specificity was 85.2 %, with PPV and NPV of 86.2 % and 83.1 %, respectively.The Point of care device resulted in a good performance for the diagnosis of low-intensity infections, it was able to identify 19 out of 25 (76 %) individuals with ⩽7 eggs, 10 out of 14 individuals (71.4 %) with 11–99 eggs and 100 % of individuals with 100–399 eggs.

Keywords: schistosomiasis, immunochromatography, naon-dot-ELISa, diagnostis device

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Authors: Supachai Supmak, Yachai Limpiyakorn

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Mutation testing can be applied for the quality assessment of test cases. Prioritization of mutation test generation has been a critical element of the industry practice that would contribute to the evaluation of test cases. The industry generally delivers the product under the condition of time to the market and thus, inevitably sacrifices software testing tasks, even though many test cases are required for software verification. This paper presents an approach of applying a social network centrality measure, PageRank, to prioritize mutation test generation. The source code with the highest values of PageRank will be focused first when developing their test cases as these modules are vulnerable to defects or anomalies which may cause the consequent defects in many other associated modules. Moreover, the approach would help identify the reducible test cases in the test suite, still maintaining the same criteria as the original number of test cases.

Keywords: software testing, mutation test, network centrality measure, test case prioritization

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Authors: Rungrawee Samawathdana, Aim-Utcha Wattanaburanon

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Test specifications for open book or notes exams provide the essential information to identify the types of the test items with validity of the evaluations process. This article explains the purpose of test specifications and illustrates how to use it to help construct the approach of open book or notes exams. The complication of the course objectives is challenging for the test designing.

Keywords: course curriculum, environment for health, internationalization, test specifications

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Authors: Ziyan Zhang

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Although the Synthetic Control Method (SCM) is now widely applied, its most commonly- used inference method, placebo test, is often problematic, especially when the treatment is not uniquely assigned. This paper discusses the problems with the placebo test under the multivariate treatment case. And, to improve the power of inferences, I further propose an Andrews-type procedure as it potentially solves some drawbacks of the placebo test. Simulations are conducted to show the Andrews’ test is often valid and powerful, compared with the placebo test.

Keywords: Synthetic Control Method, Multiple treatments, Andrews' test, placebo test

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Authors: Sidra Islam, Moin Uddin, Mir A. Rouf

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A wide variety of pathological disorders owing to hyperglycemic conditions involves structural rearrangements and condensations of proteins. The implication of methylglyoxal (MG) modified immunoglobulin G (IgG) in the onset and progression of diabetes type 2 (T2DM) is studied in the present study. Using biophysical and biochemical approaches MG was found to perturb the structure of IgG, effect its microenvironment and leads to aggregate formation. Furthermore, MG-IgG was found to be highly immunogenic inducing high titre antibodies in female rabbits. Clinical studies revealed the presence of circulating anti-MG-IgG antibodies as analyzed by direct binding ELISA. The circulating auto antibodies were highly specific for MG-IgG as revealed by inhibition ELISA. Thus it can be concluded that MG is a powerful agent with a high damaging potential. To IgG. It is highly capable of generating immune response that contributes to the immunopathology associated with diabetes. Dicarbonyl adducts may emerge as potential biomarkers for T2DM.

Keywords: immunogenicity, Immunoglobulin G, methylglyoxal, Type 2 Diabetes Mellitus

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Authors: Monika Dmitrzak-Weglarz, Aleksandra Rajewska-Rager, Maria Skibinska, Natalia Lepczynska, Piotr Sibilski, Joanna Pawlak, Pawel Kapelski, Joanna Hauser

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Epidermal growth factor (EGF) is a well-known neurotrophic factor that involves in neuronal growth and synaptic plasticity. The proteomic research provided in order to identify novel candidate biological markers for mood disorders focused on elevated EGF serum level in patients during depression episode. However, the EGF association with mood disorder spectrum among adolescents and young adults has not been studied extensively. In this study, we aim to investigate the serum levels of EGF in adolescents and young adults during hypo/manic, depressive episodes and in remission compared to healthy control group. In our study, we involved 80 patients aged 12-24 years in 2-year follow-up study with a primary diagnosis of mood disorder spectrum, and 35 healthy volunteers matched by age and gender. Diagnoses were established according to DSM-IV-TR criteria using structured clinical interviews: K-SADS for child and adolescents, and SCID for young adults. Clinical and biological evaluations were made at baseline and euthymic mood (at 3th or 6th month of treatment and after 1 and 2 years). The Young Mania Rating Scale and Hamilton Rating Scale for Depression were used for assessment. The study protocols were approved by the relevant ethics committee. Serum protein concentration was determined by Enzyme-Linked Immunosorbent Assays (ELISA) method. Human EGF (cat. no DY 236) DuoSet ELISA kit was used (R&D Systems). Serum EGF levels were analysed with following variables: age, age under 18 and above 18 years old, sex, family history of affective disorders, drug-free vs. medicated. Shapiro-Wilk test was used to test the normality of the data. The homogeneity of variance was calculated with Levene’s test. EGF levels showed non-normal distribution and the homogeneity of variance was violated. Non-parametric tests: Mann-Whitney U test, Kruskall-Wallis ANOVA, Friedman’s ANOVA, Wilcoxon signed rank test, Spearman correlation coefficient was applied in the analyses The statistical significance level was set at p<0.05. Elevated EGF level at baseline (p=0.001) and at month 24 (p=0.02) was detected in study subjects compared with controls. Increased EGF level in women at month 12 (p=0.02) compared to men in study group have been observed. Using Wilcoxon signed rank test differences in EGF levels were detected: decrease from baseline to month 3 (p=0.014) and increase comparing: month 3 vs. 24 (p=0.013); month 6 vs. 12 (p=0.021) and vs. 24 (p=0.008). EGF level at baseline was negatively correlated with depression and mania occurrence at 24 months. EGF level at 24 months was positively correlated with depression and mania occurrence at 12 months. No other correlations of EGF levels with clinical and demographical variables have been detected. The findings of the present study indicate that EGF serum level is significantly elevated in the study group of patients compared to the controls. We also observed fluctuations in EGF levels during two years of disease observation. EGF seems to be useful as an early marker for prediction of diagnosis, course of illness and treatment response in young patients during first episode od mood disorders, which requires further investigation. Grant was founded by National Science Center in Poland no 2011/03/D/NZ5/06146.

Keywords: biological marker, epidermal growth factor, mood disorders, prediction

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Authors: Serkan Yesil

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Cucurbits (the Cucurbitaceae family) include 119 genera and 825 species distributed primarily in tropical and subtropical regions of the world. The major cultivated cucurbit species such as melon (Cucumis melo L.), cucumber (Cucumis sativus L.), squash (Cucurbita pepo L.), and watermelon (Citrullus lanatus (Thunb) Matsum.&Nakai) are important vegetable crops worldwide. Squash is grown for fresh consuming, as well as its seeds are used as a snack in Turkey like some Mediterranean countries and Germany, Hungary, Austria and China. Virus diseases are one of the most destructive diseases on squash which is grown for seeds in Aksaray province. In this study, it was aimed to determine the virus infections in major squash growing areas in Aksaray province. Totally 153 plant samples with common virus symptoms like mosaic, curling, blistering, mottling, distortion, shoestring, stunting and vine decline were collected from squash plants during 2014. In this study, DAS-ELISA method is used for identifying the virus infections on the plant samples. According to the results of the DAS-ELISA 84.96 % of plant samples were infected with Zucchini yellow mosaic Potyvirus (ZYMV), Watermelon mosaic Potyvirus-2 (WMV-2), Cucumber mosaic Cucumovirus (CMV), Papaya ringspot Potyvirus-watermelon strain (PRSV-W) and Squash mosaic Comovirus (SqMV). ZYMV was predominant in the research area with the ratio of 66.01 %. WMV-2 was the second important virus disease in the survey area, it was detected on the samples at the ratio of 57.51 %. Also, mixed infections of those virus infections were detected commonly in squash. Especially, ZYMV+WMV-2 mixed infections were common. Cucumber green mottle mosaic Tobamovirus (CGMMV) was not present in the research area.

Keywords: Aksaray, DAS-ELISA, edible seed squash, WMV-2, ZYMV

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Authors: Farnaz Fotovvatikhah, Javad Akbari

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Test time of a test architecture is an important factor which depends on the architecture's delay and test patterns. Here a new architecture to store the test results based on network on chip is presented. In addition, simple analytical model is proposed to calculate link test time for built in self-tester (BIST) and external tester (Ext) in multiprocessor systems. The results extracted from the model are verified using FPGA implementation and experimental measurements. Systems consisting 16, 25, and 36 processors are implemented and simulated and test time is calculated. In addition, BIST and Ext are compared in terms of test time at different conditions such as at different number of test patterns and nodes. Using the model the maximum frequency of testing could be calculated and the test structure could be optimized for high speed testing.

Keywords: test, nano on-chip network, JTAG, modelling

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Authors: Gulyas Erzsebet

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The multimerisation of proteins is a common feature of many cellular processes; however, it could also impair protein functions and/or be associated with the occurrence of diseases. Thus, development of a research tool monitoring the appearance/presence of multimeric protein forms has great importance for a variety of research fields. Such a tool is potentially applicable in the ante-mortem diagnosis of certain conformational diseases, such as transmissible spongiform encephalopathies (TSE) and Alzheimer’s disease. These conditions are accompanied by the appearance of aggregated protein multimers, present in low concentrations in various tissues. This detection is particularly relevant for TSE where the handling of tissues derived from affected individuals and of meat products of infected animals have become an enormous health concern. Here we demonstrate the potential of such a multimer detection approach in TSE by developing a facile approach. The Biospiral-Detect system resembles a traditional sandwich ELISA, except that the capturing antibody that is attached to a solid surface and the detecting antibody is directed against the same or overlapping epitopes. As a consequence, the capturing antibody shields the epitope on the captured monomer from reacting with the detecting antibody, therefore monomers are not detected. Thus, MDS is capable of detecting only protein multimers with high specificity. We developed an alternative system as well, where RNA aptamers were employed instead of monoclonal antibodies. In order to minimize degradation, the 3' and 5' ends of the aptamer contained deoxyribonucleotides and phosphorothioate linkages. When compared the monoclonal antibodies-based system with the aptamers-based one, the former proved to be superior. Thus all subsequent experiments were conducted by employing the Biospiral -Detect modified sandwich ELISA kit. Our approach showed an order of magnitude higher sensitivity toward mulimers than monomers suggesting that this approach may become a valuable diagnostic tool for conformational diseases that are accompanied by multimerization.

Keywords: diagnosis, ELISA, Prion, TSE

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Authors: Muhammad Suhail Ibrahim, Ahmad Mujtaba

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Background: The accuracy of the most frequently used tests for diagnosing Helicobacter pylori is always under consideration in clinical settings. A reliable diagnosis is crucial to confirm the success of therapy. Objective: The aim of this research was to study the isolation frequency of H. pylori from patients compatible with gastritis or gastric ulcer and to compare some feasible non-invasive and invasive methods for the diagnosis of infection. Materials and Methods: Ninety-six gastric biopsy and blood samples were obtained with various gastroduodenal symptoms after obtaining informed consent. The biopsies were analyzed and compared using the culture, microscopic examination, histopathology, Rapid urease RUT), serology, biochemical, antibiotic susceptibility test and molecular method. Results: A number of 40 (41.67%) were considered H. pylori positive in both histopathology and RUT. On the other hand, 46 patients were positive against anti IgA and IgG by ELISA. Eighteen biopsies were positive according to the culture test. This was further confirmed by endoscopic examination, urease, catalase and oxidase tests. A high percentage of resistance to polymyxin B, amoxicillin, and kanamycin was observed (100, 88.89, and 77.78%, respectively). A gene (Cag A) was also detected by using molecular technique which appeared positive in 16 patients. The sensitivity/specificity (%) of diagnostic method was 95/77 for histology, 100/83.5 for rapid urease, 85.7/90 for gram staining, 100/66.6 for IgG serology, 100/79.5 for IgA serology, 100/75.0 for PCR, 100/79.04 for combination of RUT and IgG serology and 100/92.4 for combination of RUT, gram staining and IgG serology. Conclusion: In view of the result obtained, PCR appeared to be the most reliable test. However, higher sensitivity and specificity were also recorded for other tests. So, for more accurate results, it is advisable not to rely solely on a single method for detection.

Keywords: helicobacter pylori, isolation, detection, culture, urease, polymerase chain reaction, antibiotic susceptibility test, dyspeptic patients

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Authors: Mohammad Alkhatatbeh, Nehad Ayoub, Nizar Mhaidat, Nesreen Saadeh, Lisa Lincz

Abstract:

CD36 is involved in the development of atherosclerosis by enhancing macrophage endocytosis of oxidized-low density lipoproteins and foam cell formation. Soluble CD36 (sCD36) was found to be elevated in type 2 diabetic patients and was supposed to act as a marker of insulin resistance and atherosclerosis. In young subjects, sCD36 was associated with cardiovascular risk factors including obesity and hypertriglyceridemia. This study was conducted to further investigate the relationship between plasma sCD36 and cardiovascular risk factors among middle-aged patients with metabolic syndrome (MetS) and healthy controls. SCD36 concentrations were determined by enzyme-linked immunosorbent assays (ELISA) for 41 patients with MetS and 36 healthy controls. Data for other variables were obtained from patients' medical records. SCD36 concentrations were relatively low compared to most other studies and were not significantly different between the MetS group and controls (P-value=0.17). SCD36 was also not correlated with age, body mass index, glucose, lipid profile, serum electrolytes and blood counts. SCD36 was not significantly different between subjects with obesity, hyperglycemia, dyslipidemia, hypertension or cardiovascular disease and those without these abnormalities (P-value > 0.05). The inconsistency between results reported in this study and other studies may be unique to the study population or be a result of the lack of a reliable standardized method for determining absolute sCD36 concentrations. However, further investigations are required to assess CD36 tissue expression in the study population and to assess the accuracy of various commercially available sCD36 ELISA kits. Thus, the availability of a standardized simple sCD36 ELISA that could be performed in any basic laboratory would be more favorable to the specialized flow cytometry methods that detect CD36+ microparticles if it was to be used as a biomarker.

Keywords: metabolic syndrome, CD36, cardiovascular risk, obesity, type 2 diabetes mellitus

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Authors: Zahrasadat Hosseini, Jie Yuan

Abstract:

Point-of-care (POC) diagnostic devices based on lab-on-a-chip (LOC) technology have the potential to revolutionize medical diagnostics. However, the development of an ideal microfluidic system based on LOC technology for diagnostics purposes requires overcoming several obstacles, such as improving sensitivity, selectivity, portability, cost-effectiveness, and prototyping methods. While numerous studies have introduced technologies and systems that advance these criteria, existing systems still have limitations. Electrochemical enzyme-linked immunosorbent assay (e-ELISA) in a LOC device offers numerous advantages, including enhanced sensitivity, decreased turnaround time, minimized sample and analyte consumption, reduced cost, disposability, and suitability for miniaturization, integration, and multiplexing. In this study, we present a novel design and fabrication method for a microfluidic diagnostic platform that integrates screen-printed electrochemical carbon/silver chloride electrodes on flexible printed circuit boards with flexible, multilayer, polydimethylsiloxane (PDMS) microfluidic networks to accurately manipulate and pre-immobilize analytes for performing electrochemical enzyme-linked immunosorbent assay (e-ELISA) for multiplexed quantification of blood serum biomarkers. We further demonstrate fast, cost-effective prototyping, as well as accurate and reliable detection performance of this device for quantification of interleukin-6-spiked samples through electrochemical analytics methods. We anticipate that our invention represents a significant step towards the development of user-friendly, portable, medical-grade, POC diagnostic devices.

Keywords: lab-on-a-chip, point-of-care diagnostics, electrochemical ELISA, biomarker quantification, fast prototyping

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Authors: Zahrasadat Hosseini, Jie Yuan

Abstract:

Point-of-care (POC) diagnostic devices based on lab-on-a-chip (LOC) technology have the potential to revolutionize medical diagnostics. However, the development of an ideal microfluidic system based on LOC technology for diagnostics purposes requires overcoming several obstacles, such as improving sensitivity, selectivity, portability, cost-effectiveness, and prototyping methods. While numerous studies have introduced technologies and systems that advance these criteria, existing systems still have limitations. Electrochemical enzyme-linked immunosorbent assay (e-ELISA) in a LOC device offers numerous advantages, including enhanced sensitivity, decreased turnaround time, minimized sample and analyte consumption, reduced cost, disposability, and suitability for miniaturization, integration, and multiplexing. In this study, we present a novel design and fabrication method for a microfluidic diagnostic platform that integrates screen-printed electrochemical carbon/silver chloride electrodes on flexible printed circuit boards with flexible, multilayer, polydimethylsiloxane (PDMS) microfluidic networks to accurately manipulate and pre-immobilize analytes for performing electrochemical enzyme-linked immunosorbent assay (e-ELISA) for multiplexed quantification of blood serum biomarkers. We further demonstrate fast, cost-effective prototyping, as well as accurate and reliable detection performance of this device for quantification of interleukin-6-spiked samples through electrochemical analytics methods. We anticipate that our invention represents a significant step towards the development of user-friendly, portable, medical-grade POC diagnostic devices.

Keywords: lab-on-a-chip, point-of-care diagnostics, electrochemical ELISA, biomarker quantification, fast prototyping

Procedia PDF Downloads 85

Authors: Zheng Sun, Weiwei Cui, Xiaodong Ma, Hongxin Jin, Dongpao Hong, Jinsong Yang, Jingyi Sun

Abstract:

With the improvement of the performance and complexity of modern equipment, automatic test system (ATS) becomes widely used for condition monitoring and fault diagnosis. However, the conventional ATS mainly works in a serial mode, and lacks the ability of testing several equipments at the same time. That leads to low test efficiency and ATS redundancy. Especially for a large majority of equipment under test, the conventional ATS cannot meet the requirement of efficient testing. To reduce the support resource and increase test efficiency, we propose a method of design for the parallel test based on the embedded system in this paper. Firstly, we put forward the general framework of the parallel test system, and the system contains a central management system (CMS) and several distributed test subsystems (DTS). Then we give a detailed design of the system. For the hardware of the system, we use embedded architecture to design DTS. For the software of the system, we use test program set to improve the test adaption. By deploying the parallel test system, the time to test five devices is now equal to the time to test one device in the past. Compared with the conventional test system, the proposed test system reduces the size and improves testing efficiency. This is of great significance for equipment to be put into operation swiftly. Finally, we take an industrial control system as an example to verify the effectiveness of the proposed method. The result shows that the method is reasonable, and the efficiency is improved up to 500%.

Keywords: parallel test, embedded system, automatic test system, automatic test system (ATS), central management system, central management system (CMS), distributed test subsystems, distributed test subsystems (DTS)

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Authors: Bela Siska Afrida, Wisnu Barlianto, Desy Wulandari, Ery Olivianto

Abstract:

Background: Allergic asthma, caused by IgE-mediated allergic reactions, remains a global health issue with high morbidity and mortality rates. Immunotherapy is the only etiology-based approach to treating asthma, but no standard biomarkers have been established to evaluate the therapy’s effectiveness. This study aims to determine the correlation between the ratios of serum levels of HDM-specific IgE/total IgE and Asthma Control Test (ACT) score as a biomarker of the response to immunotherapy in pediatric allergic asthma patients. Patient and Methods: This retrospective cohort study involved 26 pediatric allergic asthma patients who underwent HDM-specific subcutaneous immunotherapy for 14 weeks at the Pediatric Allergy Immunology Outpatient Clinic at Saiful Anwar General Hospital, Malang. Serum levels of HDM-Specific IgE and Total IgE were measured before and after immunotherapy using Chemiluminescence Immunoassay and Enzyme-linked Immunosorbent Assay (ELISA) method. Changes in asthma control were assessed using the ACT score. The Wilcoxon Signed Ranked Test and Spearman correlation test were used for data analysis. Results: There were 14 boys and 12 girls with a mean age of 6.48 ± 2.54 years. The study showed a significant decrease in serum HMD-specific levels before immunotherapy [9.88 ± 5.74 kuA/L] compared to those of 14 weeks after immunotherapy [4.51 ± 3.98 kuA/L], p = 0.000. Serum Total IgE levels significant decrease before immunotherapy [207.6 ± 120.8IU/ml] compared to those of 14 weeks after immunotherapy [109.83 ± 189.39 IU/mL], p = 0.000. The ratios of serum HDM-specific IgE/total IgE levels significant decrease before immunotherapy [0.063 ± 0.05] compared to those of 14 weeks after immunotherapy [0.041 ± 0.039], p = 0.012. There was also a significant increase in ACT scores before and after immunotherapy (each 15.5 ± 1.79 and 20.96 ± 2.049, p = 0.000). The correlation test showed a weak negative correlation between the ratios of HDM-specific IgE/total IgE levels and ACT score (p = 0.034 and r = -0.29). Conclusion: In conclusion, this study showed that a decrease in HDM-specific IgE levels, total IgE levels, and HDM-specific IgE/total IgE ratios, and an increase in ACT score, was observed after 14 weeks of HDM-specific subcutaneous immunotherapy. The weak negative correlation between the HDM-specific IgE/total IgE ratio and the ACT score suggests that this ratio can serve as a potential biomarker of the effectiveness of immunotherapy in treating pediatric allergic asthma patients.

Keywords: HDM-specific IgE/total IgE ratio, ACT score, immunotherapy, allergic asthma

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Authors: Nagmani Lnu

Abstract:

Test automation is a vital requirement of any organization to release products faster to their customers. In most cases, an organization has an approach to developing automation but struggles to maintain it. It results in an increased number of Flaky Tests, reducing return on investments and stakeholders’ confidence. Challenges grow in multiple folds when automation is for UI behaviors. This paper describes the approaches taken to identify the root cause of automation instability in an extensive payments application and the best practices to address that using processes, tools, and technologies, resulting in a 75% reduction of effort.

Keywords: automation stability, test stability, Flaky Test, test quality, test automation quality

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Authors: J. Carlos Kuri, Varun Vij, Raymond J. Turner, Orly Yadid-Pecht

Abstract:

Celiac disease (CD) is an immune system disorder that is triggered by ingesting gluten. As a gluten-free (GF) diet has become a concern of many people for health reasons, a gold standard had to be nominated. Enzyme-linked immunosorbent assay (ELISA) has taken the seat of this role. However, multiple limitations were discovered, and with that, the desire for an alternative method now exists. Nucleic acid-based aptamers have become of great interest due to their selectivity, specificity, simplicity, and rapid-testing advantages. However, fluorescence-based aptasensors have been tagged as unstable, but lifespan details are rarely stated. In this work, the lifespan stability of a fluorescence-based aptasensor is shown over an 8-week-long study displaying the accuracy of the sensor and false negatives. This study follows 22 different samples, including GF and gluten-rich (GR) and soy sauce products, off-the-shelf products, and reference material from laboratories, giving a total of 836 tests. The analysis shows an accuracy of correctly classifying GF and GR products of 96.30% and 100%, respectively when the protocol is augmented with molecular sieves. The overall accuracy remains around 94% within the first four weeks and then decays to 63%.

Keywords: aptasensor, PEG, rGO, FAM, RM, ELISA

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Authors: Rostika Flora, Muhammad Zulkarnain, Syokumawena

Abstract:

Background: Aerobic physical exercises are recommended to keep body fit and healthy although physical exercises themselves can increase body metabolism and oxygen and can lead into tissue hypoxia. Oxygen pressure can serve as Vascular Endhothelial Growth Factor (VEGF) regulator. Hypoxia increases gene expression of VEGF through ascendant regulation of HIF-1. VEGF is involved in regulating angiogenesis process. Aerobic physical exercises can increase the concentration of VEGF in brain and enables angiogenesis process. We have investigated the influence of aerobic physical exercise to the VGEF concentration of wistar rat’s brain. Methods: This was experimental study using post test only control group design. Independent t-test was used as statistical test. The samples were twenty four wistar rat (Rattus Norvegicus) which were divided into four groups: group P1 (control group), group P2 (treatment group with once-a-week exercise), group P3 (treatment group with three time-a-week exercise), and group P4 (treatment group with seven time-a-week exercise). Group P2, P3, and P4 were treated with treadmil with speed of 20 m/minute for 30 minutes. The concentration of VEGF was determined by ELISA. Results: There was a significant increase of VEGF in treatment group compared with control one (<0.05). The maximum increase was found in group P2 (129.02±64.49) and the minimum increase was in group P4 (96.98±11.20). Conclusion: The frequency of aerobic physical exercises influenced the concentration of Vascular Endhothelial Growth Factor (VEGF) of brain tissue of Rattus Norvegicus.

Keywords: brain tissue, hypoxia, physical exercises, vascular endhothelial growth factor

Procedia PDF Downloads 491