Enzyme-linked Fluorescent Assay Research Articles

Healthcare-associated Diarrhea due to Clostridioides difficile in Patients Attending a Tertiary Care Teaching Hospital of North India.

Healthcare-associated diarrhea (HCAD) is diarrhea that develops at least after 3 days of hospitalization, with the most common infectious cause being Clostridioides difficile. Over the last decade, there has been a remarkable growth in the frequency and severity of C. difficile infection (CDI), making it one of the most prevalent healthcare-associated infections. This study aimed to analyze the prevalence and risk factors associated with CDI. A total of 107 patients with clinical suspicion of having HCAD were included in this study. Enzyme-linked fluorescent assay (ELFA) technique-based glutamate dehydrogenase (GDH) and toxin A/B assay were used as per the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) for diagnosing CDI. The details about associated comorbidities were retrieved from the hospital information system records. The presence of risk factors was noted. Risk factors associated with CDI were looked for. Out of the 107 stool samples received in the microbiology laboratory from patients with suspected HCAD eight (7.6%) samples were positive for CDI. The most frequent comorbidity observed in these patients was renal illness (acute or chronic kidney disease). In this study, a total of 7/8 cases were on multiple antibiotics most common being carbapenem. The 6-year prevalence of CDI observed in this study was found to be 7.6% risk factors, associated with CDI were kidney disease, diabetes mellitus, malignancy, and exposure to broad-spectrum antibiotics. Raj N, Agarwal J, Singh V, et al. Healthcare-associated Diarrhea due to Clostridioides difficile in Patients Attending a Tertiary Care Teaching Hospital of North India. Euroasian J Hepato-Gastroenterol 2024;14(1):60-64.

Seroprevalence of Toxoplasma gondii and Associated Risk Factors among Pregnant Women in Algeria.

Toxoplasma gondii is an obligate intracellular protozoan parasite affecting all warm-blooded vertebrates, including humans. Infections in humans can lead to severe clinical manifestations in pregnant women and immunocompromised patients. The aim of the present study was to assess seroprevalence of T. gondii infection and to identify the associated risk factors among pregnant women from southern Algeria. A cross-sectional study was conducted from March 2021 to May 2022 among 1,345 pregnant women. A structured questionnaire was used to obtain information on risk factors associated with infection. Participants were screened for anti T. gondii IgG and IgM antibodies using the enzyme-linked fluorescent assay. The overall IgG and IgM seroprevalence was 13.6% and 0.89%, respectively. A significant association was found between seroprevalence of anti T. gondii IgG and history of spontaneous abortion (P = 0.016). Moreover, an increasing trend of seroprevalence was detected in the autumnal season (P = 0.030), and eating raw or undercooked meat was found to be significantly associated with anti-T. gondii IgM seropositivity (P = 0.002). Multivariate analysis showed that women who had experienced spontaneous abortion, regularly ate undercooked meat, and used bottled water in summer were more likely to contract infection with T. gondii. The majority (86.4%) of the studied pregnant women were serologically negative for toxoplasmosis and hence were susceptible to primary acute infection during pregnancy and possible fetal anomalies. Therefore, health education and awareness of the disease and its transmission to women, especially during pregnancy, is imperative.

Carcinoembryonic antigen and cancer antigen 125 simultaneously determined using a fluidics-integrated dual carbon electrode

Carcinoembryonic antigen (CEA) and cancer antigen 125 (CA125) are important tumor markers for the diagnosis of ovarian cancer at the metastasis stage. Here, CEA and CA125 were simultaneously detected by a label-free electrochemical immunosensor. Using a stencil and carbon ink, a dual carbon electrode (DCE) was fabricated on a poly(vinyl chloride) substrate. Gold particles (AuPs) were electrodeposited on the electrode as a signal amplifier and support for immobilized antibodies. Linear sweep voltammetry was utilized to detect the tumor markers based on the specific binding of antigens to antibodies. Scanning electron microscopy and atomic force microscopy were used to characterize the fabricated DCE. The results were compared to those of a commercial DCE. Using the developed DCE immunosensor, we achieved a linear range of CEA from 0.5 to 25 ng mL−1 and a linear range of CA125 from 2 to 50 U mL−1. The limits of detection of CEA and CA125 were 0.15 ng mL−1 and 0.6 U mL−1, respectively. The reliability of the immunosensor was evaluated using human serum. The results were consistent with those obtained from the reference enzyme-linked fluorescence assay. The immunosensor was integrated with a small plasma membrane to form a fluidic cell (Flu-iDCE) based on acrylic sheet. The membrane had no effect on the detection of the tumor markers with the proposed Flu-iDCE. This first report of a fluidic device integrated with a DCE demonstrates the potential of the device for point-of-care detection of CEA and CA125.

Serum Level of 25-Hydroxyvitamin D in Children with Epilepsy Receiving Long-Term Antiepileptic Treatment

Background: Long-term treatment with antiepileptic drugs (AED) requires monitoring of potential side effects, one of them is the decreasing in serum vitamin D level. Low serum vitamin D level is also a global health problem in healthy children. There are conflicting results regarding the low serum vitamin D level in epileptic children due to treatment with AED. Purpose: This study aimed to compare serum vitamin D level between healthy children and children with epilepsy receiving long-term AED treatment in Soetomo General Hospital. Methods: This was a cross-sectional study conducted in the pediatric neurology clinic of Soetomo general academic hospital from August 2018 to July 2019. Subjects are children ages 2-18 years with epilepsy receiving AED treatment for over 6 months (n=22), with healthy children as control (n=22). Peripheral serum 25-hydroxyvitamin D level were measured using enzyme-linked fluorescence assay (ELFA) method. The differences of those groups were analyzed by comparing the mean of 25-hydroxyvitamin D level of both groups using Anova and T-test (95% CI). Results: Low serum vitamin D level were identified in 27% children with epilepsy and 13% healthy children. The mean of serum 25-hydroxyvitamin D level in children with epilepsy and healthy children was 21.5 ng/ml (SD 8.41) and 34.3 ng/ml (SD 10.09), respectively. There was a significant difference between groups (p<0.05). Conclusion: Children with epilepsy receiving long-term AED treatment have lower level of serum 25-hydroxyvitamin D than healthy children.

Incidence of Clostridium difficile in Patients with Antibiotic Associated Diarrhea

Background: This study was performed to determine the magnitude of Clostridioides difficile infection (CDI) in a tertiary care hospital in patients with antibiotic associated diarrhea (AAD) and to study the risk factors associated with this disease.
 Methods: A descriptive study was conducted in the department of Microbiology in a tertiary care hospital during December 2019 to May 2021. Stool samples were collected from patients with signs and symptoms of AAD who had been consuming antibiotic or anticancer drugs durng six weeks before the sampling. The samples were subjected to C. difficile glutamate dehydrogenase (GDH) enzyme and CD toxin A & B detection by Enzyme Linked Fluorescent Assay (ELFA). Patient’s demographic features and clinical details were noted and statistically correlated with the test results
 Results: Among the total 70 samples tested 20 (28%) were positive for GDH alone and 12 (17%) were positive for both GDH and CD toxin A and B. Fluoroquinolones was a significant risk factor in the study. Sepsis and colitis was found to have significant association with C.difficile infection in our study. The crude mortality rate was 17%.
 Conclusion: Prompt and precise diagnosis and knowledge about the risk factors of CDI helps in effective management and prevention of CDI.

Fetuin-A level in patients with untreated thyroid dysfunction

Fetuin-A, a plasma glycoprotein, has been demonstrated to play an essential role in the pathogene­sis of several metabolic disorders. This study aimed to estimate fetuin-A serum level in patients with newly diagnosed primary hyperthyroidism (PHT) and subclinical hypothyroidism (SCH) and to examine its correlation with thyroid hormones level, age and sex of patients. The study involved 90 patients with untreated thyroid dysfunction verified with thyroid function test (45 with PHT and 45 with SCH) and 90 control subjects. Triiodo­thyronin (T3), tetraiodothyronin (T4), and thyroid stimulating hormone (TSH) serum concentrations were measured with enzyme-linked fluorescent assay (ELFA), fetuin-A concentration was measured with enzyme-linked immunosorbent assay (ELISA). It was demonstrated that the level of fetuin-A was significantly higher in the PHT group as compared with the control group and showed a significant positive correlation with the T3 level. In the SCH group, the level of fetuin-A was significantly lower and showed a negative correlation with TSH level. Fetuin-A level rose with age in the PHT group and was unaffected by sex in all studied groups. The perfect AUC value obtained for fetuin-A in the comparison between PHT and SCH groups suggests its potential use as a reliable diagnostic marker to differentiate between these two thyroid conditions.

Comparative Diagnostic Value of Dengue Infection Using ELFA and Two Commercial Immuno-Chromatography Tests

Dengue Virus (DEN-V) can lead to a broad spectrum of clinical presentations, from mild symptoms to mortality. Based on the presence of antibodies, dengue infection is categorized into primary and secondary dengue. Early diagnosis is crucial for effective treatment of DEN-V infection. Non-Structural Antigen 1 (NS1) and dengue antibodies like immunoglobulin M (IgM) and immunoglobulin G (IgG) anti-dengue are standard assays used for dengue diagnosis. Various immunoassay methods, including Enzyme-Linked Fluorescent Assay (ELFA) and Immuno-Chromatographic Tests (ICT), are employed to detect these antigens and antibodies. This study aimed to compare the diagnostic value between ELFA and two commercial ICTs for detecting NS1 antigen, IgM/IgG anti-dengue. Seventy suspected dengue patients with fever lasting two to seven days at Premier Hospital, Surabaya were enrolled in this study. Blood serum samples from the individuals were tested for NS1 antigen and IgM/IgG anti-dengue using VIDAS® and two commercial ICTs (Boson and SD Bioline). The sensitivity of NS1 antigen assays using ELFA and two ICTs was calculated, as well as the agreement rate between ELFA and both ICTs. ELFA demonstrated high sensitivity (77.97%) for NS1 antigen detection in diagnosing dengue infection compared to two ICTs (76.27% and 45.45%). Using an automated system in ELFA can offer more excellent diagnostic value and objective results and determine the cut-off ratio of IgM/IgG antibodies. All of this comes at a cost comparable to ICT.

Plasma D-Dimer Value Corrected with Some Physiological and Inflammatory Markers (C-Reactive Protein and Ferritin) in Iraqi Patients with COVID-19 Infection

Since the beginning of the COVID-19 pandemic it has been observed that patients have elevated plasma levels of D-dimer and some inflammatory markers (ferritin, interleukine 6, C-reactive protein (CRP) or fibrinogen). Some studies point to the existence of a certain correlation between those markers and D-dimer. CRP is a protein discovered in the 1930s by Tillett and Francis and is an acute phase reactant. It is a pentameric protein which is synthesized by the liver under the action of cytokine interleukin 6 (IL-6). D-dimers are multiple peptide fragments produced as a result of degradation of crosslinked fibrin, mediated by plasmin. A total of 60 patients were recruited and categorized into :1- group1 (controls), 2- group 2 (COVID-19 patients). 5 ml of blood was obtained from each patient by vein puncture, using 5 ml disposable syringes, then centrifuged at 3000 rpm for 10 minutes to collect the serum. D- dimer, C-reactive protein were measured by using (Roche Diagnostics GmbH, Mannheim, Germany). At the same time, the Ferritin was assessed by using a miniVIDAS analyzer for the fluorescent enzymatic detection of β2-microglobulin (β2M) using the technique. Enzyme Linked Fluorescent Assay (ELFA) (BioMerieux). Our results showed that there was a non- significant difference in the P-values between control and patients males and females. The Mean ± SE of age in control group was 45.90 ± 3.34, while the Mean ± SE of age in patients was 45.35 ± 2.52. There was a non- significant difference between the two groups, the Mean ± SE of CRP in control group was 5.04 ±0.81, while the Mean ± SE of CRP in COVID-19 patients was 37.16 ±3.24, there was a highly Significant differences between them (P≤0.01). The CRP of COVID-19 patients were compared with those of control patients, the results shows significant increased CRP in covid-19 patients group as a compression with the control group, the findings of the study is similar to Jacob Lentner, etal findings. In response to infections, the liver synthesizes significant quantities of acute-phase proteins (APPs), such as CRP. This acute inflammatory protein is a highly sensitive biomarker for inflammation, tissue damage, and infection. It has been shown that CRP levels are correlated with levels of inflammation. CRP levels can promote phagocytosis and activate the complement system. In other words, CRP binds to microorganisms and promotes their removal through phagocytosis. D-dimers are one of the fragments produced when plasmin cleaves fibrin to break down clots. Our study showed that the serum D-dimer concentrations in patients significantly higher than those in control group which is similar to Mamta Soni, etal, 2020 findings. Elevated D-dimer levels have emerged as a consistent finding in severely ill COVID-19 patients, Multiple studies have identified an association between higher D-dimer levels and an increased risk of mortality in the COVID-19 patient population. Ferritin is an iron-storing protein; its serum level reflects the normal iron level and helps the diagnosis of iron deficiency anemia. Circulation ferritin level increases during viral infections and can be a marker of viral replication. Our study showed a significant increase in ferritin level in covid-19 patients group compared to control group, which consider similar to [] results. Although the exact cause for elevated ferritin in COVID-19 infection is unknown, it could be influenced by cytokine release or cellular damage that results in the leakage of intracellular ferritin. It has been previously shown that ferritin is a direct indicator of cellular damage suggestive of an association between organ damage and ferritin production. This could later cause cell death, known as ferroptosis. It is suggested that inflammation associated with sepsis could alter iron metabolism and deficiency to facilitate the immune system, which could be an early sign of COVID.

The Relationship Between D-Dimer Levels and the Value of Prothrombin Time and Activated Partial Thromboplastin Time in Patients with COVID-19 At Rsup Dr. M. Djamil Padang

Covid-19 spread widely resulting in a global pandemic that continues to this day. People who are confirmed to have severe COVID-19 will cause thrombosis which results in increased levels of D-dimer prolonging Prothrombin Time or Activated Partial Thromboplastin Time, increased fibrinogen, and thrombocytopenia. one of the parameters that determine the presence of coagulation can be seen from the levels of D-dimer, PT and APTT. The purpose of this study was to determine the relationship between D-dimer levels and the value of prothrombin time and activated partial thromboplastin time in Covid-19 patients. This research is analytic observational with a cross sectional study design. The sample in this study amounted to 30 respondents. The examination carried out was D-dimer levels using the ELFA (Enzyme Linked Fluorescent Assay) method using Vidas, Prothrombin Time and Activated Partial Thromboplastin Time using the Flouroscence Flowcytometry method using the Sysmex Cs 2500 tool. The data were analyzed using the Spearman’s rho correlation test. The results of the study on Covid-19 average levels of D-dimer 1696.23 g/ml ±1567.448 SD, the value of Prothrombin Time 11.103±1.1898 seconds and Activated Partial Thromboplastin Time 32.643±17.3796 seconds. Conclusion There is a significant relationship between D-dimer levels with the Prothrombin Time P value of 0.003 and with Prothrombin Time and Activated Partial Tromboplastin Time P value of 0.031. There was no significant relationship between D-dimer levels and Activated Partial Tromboplastin Time P value 0.110. High D-dimer levels are accompanied by a prolonged prothrombin time.

Comparison of ECL, ELISA and ELFA immunoassays

The aim of these study was the evaluation of electrochemiluminescence assay (ECL), enzyme-linked immunosorbent assay (ELISA) and enzyme-linked fluorescent assay (ELFA), for an early diagnosis of Cytomegalovirus infections in pregnant women. In medical diagnostics, it is necessary to determine the most sensitive techniques for the detection of this pathogen, due to it’s multiple fetal infections during pregnancy. The ECL technique resulted with the highest sensitivity and specificity values. However, for diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history and other clinical examinations.

Comparison of three immunoassays for detection of anti-toxoplasma IgM antibodies in pregnant women

Abstract Aim of investigation Comparison of an electrochemiluminescence technique (ECL, applied in Cobas 6000 instrument), an enzyme-linked immunosorbent assay (ELISA, applied in CHORUS instrument) and an enzyme-linked fluorescent assay (ELFA, applied in MINI-VIDAS instrument) for early diagnosis of Toxoplasma gondii infection in pregnant women. Medical diagnosis is required to determine the most sensitive techniques to diagnose this pathogen, in the framework of which is developed this scientific work. This is very important due to the multiple fetal infections during pregnancy. Methods ECL, ELISA and ELFA techniques are used to detect anti-toxoplasma IgM antibodies in pregnant women, during the first trimester of pregnancy. 200 samples were analyzed with each technique and sensitivity and specificity are evaluated for each of them. Results ECL technique has resulted in higher sensitivity (100%) and specificity (99.3%), while ELISA technique has resulted in lower sensitivity (87.5%) and specificity (97.3%). Conclusion Analysis of the results confirmed the usefulness of ECL technique for an early diagnosis of Toxoplasma gondii infections in pregnant women. Anyway, for diagnostic purposes, the results should always be assessed in conjuction with the patient’s medical history and other clinical examinations.

Serum Copper, Iron, and Total Iron Binding Capacity in Hypothyroidism: A Case Control Study

Background: Thyroid hormone metabolism is linked to iron metabolism. Thyroperoxidase, a key enzyme in the biosynthesis of thyroid hormones, is iron-dependent. Thus, iron deficiency might be the primary cause of hypothyroidism. Copper is another trace element that has been linked to thyroid status. Copper regulates excessive thyroxine (T4) absorption and reduces cell damage during thyroid hormone synthesis. The present study clarified the possible correlations between iron and copper levels as well as total iron binding capacity (TIBC) and triiodothyronine (T3), T4, as well as thyroid-stimulating hormone (TSH) levels in healthy and hypothyroid subjects.Materials and methods: Thirty-five healthy subjects and 35 hypothyroid subjects were included in this study. Serum T3, T4, and TSH levels were measured using the enzyme linked fluorescence assay. Serum iron levels and TIBC were estimated using ferrozine/magnesium carbonate method, while serum copper levels were estimated using colorimetric method.Results: Copper levels were not significantly different between healthy and hypothyroid subjects. Iron and T4 levels were significantly lower in hypothyroid subjects compared with those in healthy subjects, while TIBC and TSH levels were significantly higher. There was no significant correlation between copper levels and T3, T4, and TSH levels.Conclusion: There were inverse correlations between TIBC and T4 as well as iron levels, and there was no significant correlation between copper levels and all thyroid function parameters. Routine examination of iron levels and thyroid function is highly recommended for early diagnosis and therapy of hypothyroidism.Keywords: total iron binding capacity, iron, copper, hypothyroidism

Correlation of serum ferritin with parathyroid hormone level in patients with transfusion-dependent thalassemia

Introduction: Thalassemia is a rare hereditary condition that causes hemoglobin synthesis deficiency. Recurrent blood transfusion in transfusion-dependent thalassemia (TDT) individuals results in iron overload and deposition in body tissues, leading to endocrine malfunctions, including parathyroid. This study aimed to determine the correlation between the level of serum ferritin and parathyroid hormone level in TDT patients. Methods: A cross-sectional study was conducted among adult TDT patients receiving blood transfusions at Dr. Soetomo General Academic Hospital, Surabaya, Indonesia from January to June 2023. Serum ferritin was measured using an enzyme-linked fluorescence assay (ELFA), while parathyroid hormone was quantified using an enzyme-linked immunosorbent assay (ELISA). The Spearman correlation analysis was used to identify the correlation between the level of serum ferritin with parathyroid hormone. Results: A total of 46 TDT patients were enrolled in the study, with a median age of 25 years old and represented predominantly by males (56.5%). The median levels of serum ferritin and parathyroid hormone were 5,672.90 ng/mL (min-max: 596.80–24,014.58 ng/mL) and 29.80 pg/mL (min-max: 10.61–97.01 pg/mL), respectively. The Spearman correlation analysis suggested a statistically insignificant negative correlation between serum ferritin and parathyroid hormone in TDT patients (r = -0.166; p=0.270). Conclusion: Serum ferritin was inversely, but not significantly, correlated with parathyroid hormone in TDT patients, suggesting that impaired parathyroid function might be associated with the increment of plasma iron.

Evaluation of the Diagnostic Performances of the SD-Bioline®HBeAg Rapid Test Used Routinely for the Management of HBV-Infected Individuals in Burkina Faso.

Hepatitis B e antigen (HBeAg) is a marker of wild-type hepatitis B virus replication. In resource-limited countries where access to enzyme-linked immunosorbent assay (ELISA) remains a challenge, rapid diagnostic tests (RDT) constitute a good alternative. The HBeAg status is employed to evaluate eligibility for antiviral therapy and to prevent the transmission of hepatitis B from mother to child (PMTCT). The objective of this study was to assess the diagnostic performance of the SD-Bioline®HBeAg RDT commonly used for detecting HBeAg in laboratories in Burkina Faso. The sample panel used was collected from HBsAg-positive patients received in the laboratory for the detection of HBeAg with the rapid test. The samples were retested for HBeAg using the VIDAS HBe/Anti-HBe enzyme-linked fluorescent assay (ELFA) (Gold standard). Then, the viral load (VL) of HBV DNA was determined using the GENERIC HBV CHARGE VIRLAE kit (GHBV-CV). The diagnostic performances of the SD-Bioline®HBeAg and its agreement with the gold standard were calculated with their 95% confidence intervals. Overall, 340 sera obtained from HBsAg-positive patients were included in this evaluation Compared to the VIDAS HBe/Anti-HBe ELFA test, the sensitivity (Se) and specificity (Sp) of the SD-Bioline®HBeAg test were 33.3% and 97.9%, respectively. The concordance between the two tests was 0.42. Depending on the viral load, the Se and Sp varied from 8.8% and 98.3% for a VL < 2000 IU/mL to 35.5% and 98.4% for a VL > 2,000,000 IU/mL. The results showed a low sensibility of the SD-Bioline®HBeAg RDT test, indicating that its use is inappropriate for the clinical management of HBV-infected patients. They also highlight the urgent need to develop HBeAg rapid tests with better sensitivities.

Analysis of total antibody levels in university hospitals health workers vaccinated against COVID-19 in Abidjan (Côte d’Ivoire)

Discovered in China in December 2019, coronavirus disease-19 (COVID-19) has confronted the world with an unprecedented crisis. Healthcare workers, the first line of defense against this pandemic, have been severely affected. Clinical trial results of the emergency vaccines showed that they all produced IgG antibodies against severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) with high rates of seroconversion. While immunization against natural challenge (COVID-19 infection) and artificial challenge (vaccination) in health care workers is relatively well described in the West, the issue is not well understood in Sub-Saharan Africa, particularly in Côte d’Ivoire, where populations are genetically distinct from Caucasians. Our aim was to investigate the magnitude of post-vaccination IgG responses to SARS-CoV-2 in healthcare workers in our African epigenetic context. A cross-sectional, multicenter, analytical study was conducted from March to May 2022 among health workers employed at the University Hospital of Abidjan and vaccinated against COVID-19. The study included 77 health workers. IgG immunoassays were performed with an enzyme-linked fluorescent assays. Data were analyzed using SPSS version 22.0 software, with a p-value ˂ 0.05 considered as a significant difference. All enrolled subjects developed anti-SRAS-Cov-2 IgG, of which 88.3% had a strong response (titer ≥ 250 Binding Antibody Units/ml). IgG titers varied significantly by gender (p=0.04). Vaccine type and number of doses did not affect IgG titers. However, a history of COVID-19 infection was associated with a 5-fold greater likelihood of developing a strong IgG response after vaccination. In conclusion, humoral IgG responses developed after vaccination against SARS-CoV-2 were robust and would be influenced by a variety of factors..

Effects of Progesterone and Other Gonadal Hormones on Glutamatergic Circuits in the Retina.

Gonadal hormones function in the retina; however, their targets have not yet been identified. Therefore, the present study examined the effects of progesterone and other gonadal hormones on glutamatergic circuits in the retina. Extracellular glutamate concentrations, which correspond to the amount of glutamate released, were examined using an enzyme-linked fluorescent assay system. The activity of glutamatergic synapses between bipolar cells and ganglion cells was investigated using a patch clamp technique. Changes in retinal thickness during pregnancy were assessed using optical coherence tomography (OCT) images. Progesterone and pregnenolone sulfate increased extracellular glutamate concentrations, whereas estrogen and testosterone did not. Progesterone increased the activity of glutamatergic synapses between bipolar cells and ganglion cells. A temporal decrease in the thickness of the peripheral retina was observed in the 1st trimester. Progesterone, but not estrogen or testosterone, activated glutamate release in the mouse retina. Increases in the concentration of progesterone during pregnancy did not induce any detectable change in retinal thickness.

Application of Enzyme-Linked Fluorescence Assay (ELFA) to Obtain In Vivo Matured Dog Oocytes through the Assessment of Progesterone Level.

Successful dog cloning requires a sufficient number of in vivo matured oocytes as recipient oocytes for reconstructing embryos. The accurate prediction of the ovulation day in estrus bitches is critical for collecting mature oocytes. Traditionally, a specific serum progesterone (P4) range in the radioimmunoassay (RIA) system has been used for the prediction of ovulation. In this study, we investigated the use of an enzyme-linked fluorescence assay (ELFA) system for the measurement of P4. Serum samples of estrus bitches were analyzed using both RIA and ELFA, and the measured P4 values of ELFA were sorted into 11 groups based on the standard concentration measured in RIA and compared. In addition, to examine the tendency of changes in the P4 values in each system, the P4 values on ovulation day (from D - 6 to D + 1) in both systems were compared. The ELFA range of 5.0-12.0 ng/mL was derived from the RIA standard range of 4.0-8.0 ng/mL. The rates of acquired matured oocytes in RIA and ELFA were 55.47% and 65.19%, respectively. The ELFA system successfully produced cloned puppies after the transfer of the reconstructed cloned oocytes. Our findings suggest that the ELFA system is suitable for obtaining in vivo matured oocytes for dog cloning.

Thyroid Hormones and Autoantibodies in Iraqi Hyperthyroid Patients

Hyperthyroidism can be defined as the status of over production of thyroid hormones: tri-iodothyronine (T3) and thyroxine (T4) as a results of thyroid gland dysfunction. The aim of the current study was to investigate the association between the abnormalities of thyroid hormones (T3 and T4) and serum levels of thyroid auto-antibodies: anti-thyroid peroxidase (TPO-Ab) and anti-thyroid stimulating hormone receptor (TR-Ab) and the state of hyperthyroidism. Blood samples were collected from 75 Iraqi individualswere early diagnosed as hyperthyroidism patients in addition to 25 Iraqi individuals as a healthy control group. Enzyme Linked Fluorescent Assay (ELFA) used to determine the levels of thyroid hormones (T3 and T4) and the level of thyroid stimulating hormone (TSH), Enzyme Linked Immuno-Sorbent Assay (ELISA) was used to determine the levels of auto-antibodies (TPO-Ab and TR-Ab). The results show that T3 levels and T4 levels in hyperthyroidism patients (2.07 nmol/L and 14.02 μg/dl respectively) increased significantly (푃&lt;0.01)compared to healthy control levels (1.33 nmol/L and 6.4 μg/dl respectively). While the levels of thyroid stimulating hormone (TSH) in hyperthyroidism patient (0.17 μIU/Ml) decreased significantly (푃&lt;0.01)when compared with healthycontrol group (1.84 μIU/Ml). The results also showed that levels of TPO-Ab and TR-Ab in hyperthyroidism patients (235.29 IU/mL and 32.05 IU/mL respectively) increased significantly (푃&lt;0.01)when compared to healthy control group (20.76 IU/mL and 0.3 IU/mL respectively). This study revealed that there is an association between thyroid hormones disorders and the positivity of thyroid auto-antibodies in hyperthyroidism patient

English

Toxoplasma gondii is an obligatory intracellular parasite that causes a zoonotic disease capable of infecting nearly all warm-blooded hosts, including humans. However, reports on the molecular prevalence of T. gondii in humans are rare in Gabon. The present study aimed to evaluate the serological and molecular prevalence of T. gondii among apparently healthy rural populations in four regions of Gabon. This study included six hundred blood samples from the Interdisciplinary Center for Medical Research (CIRMF) bank, including 300 women and 300 men living in 111 villages. Blood samples were screened using enzyme-linked fluorescent assay (ELFA), while buffy coat samples were analyzed using PCR analyses. Of the 600 samples screened, 548 (91.3%) showed IgG antibodies against T. gondii; 11 (2%) had both IgG and IgM. Among the 548 positive samples, 155 (28%) had higher IgG titers (&gt;300 UI/ml), and 49 of them (31.6%) were detected with T. gondii DNA. The present findings on human toxoplasmosis in Gabon suggest that at an older age, reactivation of old infections seems more frequent than new infections, as indicated by the presence of T. gondii using PCR among elevated IgG subjects without IgM. Further studies should be performed to identify the genotypes of T. gondii that infect humans in Gabon. Key words: Toxoplasma gondii, humans, DNA, Gabon.

Physiological effects of prolactin on early pregnancy loss

Abortion generally presented as two types; the spontaneous and the induced abortion. Spontaneous one is the termination of pregnancy before fetal viability which occurs spontaneously. About 80% of spontaneous abortion losses occur in the first trimester, the incidence decreases with advance in pregnancy. Prolactin is a polypeptide hormone that is synthesized in and secreted from specialized cells of the anterior pituitary gland, the lactotrophs. The major action of prolactin is lactogenesis (milk production). It is the major hormone stimulating the production of milk. Prolactin seems to play an important role in implantation and subsequent placentation in the human endometrium, and it is recognized as a crucial signal for the initiation and maintenance of decidualization. In addition to that, prolactin inhibits the secretion of gonadotropin releasing hormone by the hypothalamus, and this inhibition lead to decreased secretion of luteinizing hormone (LH), which normally initiates ovulation, therefore with hyperprolactinemic state ovulation, does not occur (anovulation). The aim of this study is to study the effects of prolactin hormone in the incidence of abortion cases in first trimester pregnancy in women in Tikrit city. Serum Prolactin was measured by using kit made in France. VIDAS Prolactin (Made in Italy), for the enzyme immunoassay determination of human prolactin in human serum or plasma by using the technique of Enzyme Linked Fluorescent Assay (ELFA). One hundred and thirty four pregnant women were participated &amp; finished the whole study; 56 normal pregnant women in first trimester as a control group and 78 women with different types of abortion in first trimester who are diagnosed by clinical examination and/or by ultrasonography as aborted group. Seventy eight aborted women were participating in the present study. Threatened abortion was the highest one among aborted women (44.9%). Followed by missed abortion (19.23%), incomplete abortion (14.1%), complete (9%), inevitable (7.69%) &amp; the last is recurrent (2.57%). Regarding serum prolactin, there is a significant difference between normal pregnant women (47.1 ± 25.4 ng/ml) as compare with aborted women (69.4 ± 48.8 ng/ml). In another word, aborted women have a serum prolactin concentration than that of normal pregnant women (p≤0.01).