Enzyme-linked Fluorescent Immunoassay Research Articles

Biochemical pattern and prevalence of thyroid disorders among adults in a tertiary hospital in North-East Nigeria

Thyroid disorders are a relatively common group of endocrine disorders globally. Thyroid function tests are critical for diagnosing, screening, and prognosticating thyroidal and non-thyroidal diseases, and their accurate interpretation is required for effective and accurate patient management. A retrospective analysis of thyroid function tests - Thyroid Stimulating Hormone (TSH), Free Thyroxine (fT4), and Free Triiodothyronine (fT3) - was carried out at a Nigerian tertiary hospital between January 2017 and January 2024. The tests were conducted using an enzyme-linked fluorescent immunoassay system and interpreted following standard protocols. A preponderance of female patients was investigated for thyroid diseases, with a male-female ratio of 2.2:1. The majority (80.8%) of the analyzed requests showed a euthyroid pattern; primary hyperthyroidism was the most common biochemical pattern of thyroid dysfunction observed, accounting for 10.8% of all the analyzed requests. Secondary hyperthyroidism and hypothyroidism were generally rare. Occasional cases (0.3%) of T3 toxicosis were also observed. Goiters remain a common presentation of thyroid diseases in our environment. Primary hyperthyroidism and primary hypothyroidism are the most common forms of thyroid dysfunction observed.

Detection of Toxoplasmosis in Pregnant Women’s In Sabratha City of Using VIDS

Toxoplasmosis is usually asymptomatic, but can have severe consequences if It occurs in immunodeficient subject or fetuses. The diagnosis of toxoplasmosis during pregnancy is often based on maternal serological testing for IgM and IgG Anti- Toxoplasma antibodies. Persistence of IgM for long periods, posses' problems in distinguishing acute from chronic infection. The evaluation of specific IgG avidity enables more accurate dating, since avidity rises progressively during the course of infection. Test that measures immunoglobulin G (IgG) is used to determine if a person has been infected. If it is necessary to try to estimate the time of infection, which is of particular importance for pregnant women, a test which measures immunoglobulin M (IgM) is also used along with other tests such as an avidity test. Toxoplasma-specific IgG antibodies are detectable 1-3 weeks after infection and remain detectable for the life of the individual. Toxoplasma-specific IgM antibodies are also detectable 1-3 weeks after infection but generally decline to nil by one year after infection. The VIDAS® TOXO IgM (TXM) assay is intended for use on the instruments of the VIDAS family (VITEK® Immunodiagnostic Assay System) as an automated enzyme-linked fluorescent immunoassay (ELFA) for the presumptive qualitative detection of anti-Toxoplasma gondii IgM antibodies in human serum. Seventy-six (76) women in the first 16 weeks of pregnancy were tested for VIDAS IgM, IgG antibodies and VIDAS Toxo-IgG avidity. Low avidity antibodies were demonstrated in 2 (33.3%) of 6 sera positive with IgM assay and 4 (12.12%) of sera positive with IgG assay. Low avidity was also detected in 2 (3.27%) of 61 sera negative with IgM. The low avidity suggesting a recent infection, while high avidity in 3 (50%) of the 6 positive IgM and 24 (72.72%) of 33 positive IgG indicating that the Infection acquired in the distant past. These findings highlight the value of VIDAS IgG avidity when used in combination with the VIDAS IgM and IgG assays to provide a confirmatory evidence of an acute infection with a single serum specimen for pregnant women.

Differences of Hepatitis B Serological Tests in Cirrhosis and Hepatocellular Carcinoma Patients

Cirrhosis and hepatocellular carcinoma (HCC) are the most common complications of chronic hepatitis B. Hepatitis B Virus (HBV) reactivation occurs in the inactive phase, characterized by reappearance of HBsAg or negative HBsAg. The prevalence of occult hepatitis B infection in cirrhosis and HCC ranges from 20% to 60%. This study aimed to analyze the differences in hepatitis B serological tests in patients with cirrhosis and HCC for diagnosis of acute or chronic hepatitis B. The current research was analytical and descriptive with a cross-sectional study design. This study involved 177 patients, including 50 cirrhosis patients and 127 HCC patients. Hepatitis B serological tests (HBsAg, HBeAg, anti-HBe, total anti-HBc) were analyzed using VIDAS instruments by the Enzyme-Linked Fluorescent Immunoassay (ELFA) method. Hepatitis B serological test results were grouped based on the interpretation of serological test results. The data were processed with the statistical test Kolmogorov-Smirnov test, independent T-test, and Chi-Square, and results with p<0.05 were reported as significant. This study found that most males suffer from cirrhosis and HCC due to chronic HBV infection, with a mean age of 57. Chronic hepatitis was most common in patients with cirrhosis and HCC (71.2%). There was no significant difference in the interpretation of the hepatitis B serological test between patients with cirrhosis and HCC, with a p-value of 0.230 (p > 0.05). There was no significant difference in interpreting the hepatitis B serological test between cirrhosis and HCC. Both reactive HBsAg and non-reactive HBsAg can be obtained in cirrhosis and HCC.

EFFECTS OF DHIKR ON SPIRITUALITY AND CORTISOL LEVELS AMONG PATIENTS WITH HEART FAILURE: A PILOT STUDY

Introduction: Depression and anxiety are common comorbid conditions in patients with heart failure (HF). Spirituality was found to have a positive impact on patients’ mental health status. However, whether the spiritual practice affects physiological modulation is still questionable. Therefore, this study aimed to analyze the effect of dhikr as spiritual practice on spirituality and cortisol levels among patients with HF. Methods: A quasi-experiment was used in this study. A sample size of 18 respondents consists of 9 respondents of the control group and 9 respondents of the treatment group. Dhikr intervention was conducted in three meetings with an interval of two days during patients’ hospitalization. Spirituality was measured using a spirituality questionnaire, whereas the cortisol levels with venous blood sampling, measured by the enzyme-linked fluorescent immunoassay (ELFA) method. Statistics analysis in this study was using t-Test for spirituality, Mann Whitney and Wilcoxon Sign Rank Test for cortisol level with a significance level of α<0.05. Results: The results showed that dhikr improves the spirituality of HF patients (p=0.000) and decreases the level of cortisol (p=0.015) significantly. Dhikr interventions appears to strengthen spirituality in patients with HF by improving stress perception which further modulates the adaptive stress response in the form of decreasing cortisol levels. Conclusions: Dhikr could be one of the spiritual nursing interventions to improve spirituality and stress hormonal modulation in patients with HF.

Analysis and identification of suspected snake venom samples using nano-ultra-high performance liquid chromatography-high resolution mass spectrometry

Snake venom is a complex mixture secreted from the glands of poisonous snakes， which contains proteins， peptides， lipids， nucleosides， sugars， amino acids， amines， metal ions， and other components. According to the toxicological classification， snake venoms can be classified as neurotoxins， anticoagulants and procoagulant toxins， cardiac toxins， other toxin proteins， and enzymes. Proteins and peptides are the key components of snake venom. The establishment of rapid， accurate analysis and identification methods for proteins in snake venom is a prerequisite for snake venom-related forensic identification， intoxication events， and pharmaceutical development. Until now， the classical analysis and identification methods have mainly been biochemical or immunoassays for DNA or proteins， such as polymerase chain reaction， agglutination test， enzyme-linked immunosorbent assay， fluorescent immunoassay， and various biosensing approaches. These methods have some limitations such as a high false-positive ratio， low sensitivity， poor anti-interference ability， and limited species discrimination capability. In recent years， with the rapid development of mass spectrometry （MS） techniques， the proteomics of snake venom has also attracted much attention and has contributed to the identification of snake species， in which non-targeted and targeted proteomics represent two main divisions. However， species identification via proteomics is in its infancy in forensic science. First， the tandem MS spectra of peptide sequences are highly complex， which poses a great challenge for the strict and accurate matching of peptides based on the rational speculation of MS fragmentation rules and theoretical calculations in non-targeted proteomics. Second， for the confirmation and identification of unknown substances， reference substances are commonly needed， but those for snake venom are lacking. Proteomics in snake venom identification is still in progress to improve the identification confidence and clarify the identification rules. In this work， a method based on nano-ultra-high performance liquid chromatography-quadrupole-orbitrap high-resolution mass spectrometry （Nano LC-MS/HRMS） and size exclusion chromatography （SEC） was developed for identifying proteins and their source species， with strict rules for five suspected snake venom samples and their contamination in one case. Three SEC elution peaks were obtained from each of the five samples， which were lyophilized and treated with trypsin in solution， and then separated and analyzed by Nano LC-MS/HRMS. First， the Full MS/dd MS2 mode was used for the non-targeted acquisition of peptide information in the samples， and after submission to the Swiss-Prot database， the protein databases of Serpentes， Colubroidea， Elapidae， Elapinae， and Naja were contracted stepwise and compared. A total of 32 proteins from Naja atra were identified under the conditions of both peptide spectrum match and false discovery rate less than 1%， and number of characteristic peptides greater than or equal to two. All of these were derived from ten families of Naja atra， mainly three-finger toxins， metalloproteinases， and phospholipase A2. Proteins D3TTC2， D5LMJ3， Q7T1K6， Q9DEQ3， and Q9YGI4 were the most common among the five samples. Finally， the parallel reaction monitoring mode was adopted to select two unique peptides for each protein for targeted verification. It was considered that a protein in the samples was truly identified when it met the strict standard "the Δm/z of at least 75% y+ and b+ ions of each unique peptide was less than 5 ppm". After these consequently procedures， we identified that all five samples contained the venom of the Naja atra. Our identification method is a systematic and strict example that can provide effective technical support for the forensic identification of snake venom poisoning， as well as for pharmaceutical development toward snake venoms.

Correlation of Triglyceride/HDL-Cholesterol Ratio and Visceral Adiposity Index with 25(OH)D in Obese Female

Obesity is a condition of excess body fat mass with cardiometabolic complications. The levels of serum 25-hydroxyvitamin D (25(OH)D) decrease in obesity because it is stored in adipose tissue compartments and is related to dyslipidemia conditions. A high triglyceride/HDL-cholesterol (TG/HDL-C) ratio in obesity is related to dyslipidemia. Visceral Adiposity Index (VAI) is one of the parameters indicating central obesity related to visceral fat distribution in dyslipidemia conditions. This study aimed to prove a correlation between TG/HDL-C ratio and VAI with 25(OH)D levels in obese females. Observational study with cross-sectional design in 66 female patients. HDL-C examinations and triglyceride using the enzymatic colorimetry method. The TG/HDL-C ratio was calculated by dividing TG by HDL. The equation obtained visceral adiposity index (WC/(36.58+(1.89xBMI)) x (TG/0.81) x (1.52/HDL). 25(OH)D examinations used the Enzyme Linked Fluorescent Immunoassay (ELFA) method. Relationship status used the spearman rank test (p<0.05). Median 25(OH) levels were 9.75 (8-18.6)ng/mL. There was a weak negative correlation between TG/HDL-C ratio with 25(OH)D levels in obese females (p=0.020; r= -0.287) and VAI with 25(OH)D in obese females (p=0.019; r= 0.287). There was a weak negative correlation between TG/HDL ratio and VAI with 25(OH)D in obese female patients.

Role of <i>Toxoplasma gondii</i> in thyroiditis in pregnant women

Toxoplasmosis (acute and latent) is the most prevalent parasitic infection worldwide and can be associated with some problems in pregnant women. Thyroid diseases are the most common endocrine disorders secondary to diabetes among pregnant women. Previous studies proposed a relationship between latent еoxoplasmosis (LT) and autoimmune thyroiditis diseases (AITDs). This study intended to investigate the frequency and correlation between toxoplasmosis and AITD in pregnant women. In this cross-sectional study, the statistical population included 1248 pregnant women at the gestational age of 916 weeks and in Tehran. The Toxoplasma IgM and IgG tests were assessed with enzyme-linked immunosorbent assay (ELISA). The diagnostic criteria for toxoplasmosis were abnormal IgG and IgM titers. In addition, FT4, TPO Ab, and TSH were evaluated using enzyme-linked fluorescence immunoassay (ELFA). TPO Ab was used to distinguish thyroid patients with autoimmune origin from those with other thyroiditis diseases. The analysis showed no significant relationship between keeping a house cat and acute toxoplasmosis. Acute and latent toxoplasmosis represented 3.4% and 29.6%, respectively. The frequency of thyroid diseases was 18.8% (hypothyroidism 15.8% versus hyperthyroidism 3%). The frequency of autoimmune thyroiditis diseases (AITDs) was 5.5%, and 27.9% of subjects with latent toxoplasmosis (LT) had a thyroiditis disease, but 13.8% of pregnant women with LT had only AITD with a significant correlation (p 0.001). Results show that Toxoplasma IgG+ can increase the risk of AITD by 10.39-fold and a higher TPO Ab titer in people with LT. It seems Toxoplasma gondii may cause thyroiditis in pregnant women likely because antigenic similarity of Toxoplasma and thyroperoxidase leads to cross-reactivity in the immune system, potentially causing AITD. It might be said that the high prevalence of LT among pregnant women may have a potential role in the stimulation of the immune system to the development of autoimmune diseases, such as AITD. So future studies could be conducted with a focus on discovering molecular similarities between thyroperoxidase and Toxoplasma antigens.

Characterization of Rotavirus Infection in Hospitalized Children under 5 with Acute Gastroenteritis 5 Years after Introducing the Rotavirus Vaccines in South Korea.

We herein characterized rotavirus infection in hospitalized children under 5 years of age with gastroenteritis after introducing rotavirus vaccines in South Korea from 20 February 2012, to 31 March 2013. Enzyme-linked fluorescent immunoassay was performed to detect rotavirus antigens. G and P genotyping was performed using nested multiplex PCR. For the failed PCR samples, sequencing was conducted. We performed a test-negative case-control study to estimate vaccine effectiveness. Vaccine effectiveness was measured using a multivariate logistic regression model. Rotavirus was detected in 16 (13.2%) of the 121 patients, with a seasonal peak in April 2012. The dominant genotypes detected were G3P[8] (33.3%) and G4P[6] (26.7%), and vaccine effectiveness against rotavirus hospitalization was 84.9% [95% CI: 23.2-97.0] in the complete vaccinated group. A higher prevalence of rotavirus infection was observed among children with siblings than those without siblings (p < 0.001). Also, the presence of siblings was significantly associated with a history of nonvaccination (p < 0.001). In conclusion, the prevalence of rotavirus followed a decreasing trend, and there was no evidence of emergences of nonvaccine-type strains. Vaccine effectiveness against rotavirus hospitalization was 84.9%. Although children with siblings were more susceptible to rotavirus infection, they were less likely to receive vaccination against rotavirus.

Utilization of a Novel Immunofluorescence Instrument Prototype for the Determination of the Herbicide Glyphosate.

An enzyme-linked fluorescent immunoassay (ELFIA) method has been developed for the quantitative analytical determination of the herbicide active ingredient glyphosate in environmental matrices (surface water, soil, and plant tissues). Glyphosate, as a ubiquitous agricultural pollutant, is a xenobiotic substance with exposure in aquatic and terrestrial ecosystems due its extremely high worldwide application rate. The immunoassay developed in Project Aquafluosense is part of a fluorescence-based instrumentation setup for the in situ determination of several characteristic water quality parameters. The 96-well microplate-based competitive immunoassay method applies fluorescence signal detection in the concentration range of 0–100 ng/mL glyphosate. Application of the fluorescent signal provides a limit of detection of 0.09 ng/mL, which is 2.5-fold lower than that obtained with a visual absorbance signal. Beside the improved limit of detection, determination by fluorescence provided a wider and steeper dynamic range for glyphosate detection. No matrix effect appeared for the undiluted surface water samples, while plant tissues and soil samples required dilution rates of 1:10 and 1:100, respectively. No cross-reaction was determined with the main metabolite of glyphosate, N-aminomethylphosphonic acid, and related compounds.

Frequency of natural antibodies and concordance analysis for anti-TOXOPLASMA IgM tests in Colombian sera of pregnant women

Interpretation of IgM anti-Toxoplasma can be problematic given the phenomena of “natural” IgM. We analyzed 1,503 sera obtained during prenatal care program, and we established natural and false-positive results by doing follow-up. In 101 samples the concordance between enzyme linked immunosorbent assay (ELISA) and two semi-automatized systems: electro-chemiluminescence immunoassay (ELECSYS) and vitek immunodiagnostic assay system (VIDAS) was calculated. In 1,503 serum, 71 (4,7%) had ELISA IgG negative and ELISA IgM positive results and in 44 of these had a second sample 4 weeks later. In second samples, 27 (61,3 %) were IgM and IgG negative (false positive result in the first sample) and 13 (29,5%) were ELISA IgM positive and IgG negative (natural IgM). ELISA assay had a poor concordance with enzyme-linked fluorescent immunoassay as well as ELECSYS tests, contrarily, enzyme-linked fluorescent immunoassay and ELECSYS had optimal concordance, with 100 of 101 sera obtaining the same result by both tests. We recommended to use automatized assays to measure IgM anti-Toxoplasma.

Diagnostic value of different interleukins and procalcitonin in critically ill patients admitted with suspected sepsis.

: Many biomarkers have now been studied such as C-reactive Protein (CRP), procalcitonin (PCT), etc., and are widely used for the diagnosis of sepsis in clinical practice which may determine the appropriate antibiotic treatment. A flowcytometric cytokine bead array (CBA) assay has now been used to determine multiple interleukins (IL), simultaneously. The aim of this study was to determine the cytokine (IL2, IL4, IL6, IL10, TNFα, INFγ, and IL17) profiles of interleukins in plasma of sepsis patients by using multiplex Flowcytometric CBA array assay. s: A total of 99 consecutive patients admitted with the suspected sepsis were studied. PCT concentrations were measured by using the enzyme-linked fluorescent immunoassay (ELFA) technique and flow cytometry-based BD™ CBA Cytokine Kit was used to evaluate levels of 7 cytokines [IL-2, IL-4, IL-6, IL-10, Tumour Necrosis Factor (TNF), Interferon- γ (IFN-γ), and IL-17A]. Microbiologically defined infection (MDI) demonstrated a positive culture report in 79/99 (79.7%) of patients. The IL6 [1873.7 (4-5000)] and IL10 [(154.7 (0-1764)] levels were significantly higher in septic patients than those in the negative MDI IL6 [901 (4-5000)] and IL10 [110.4 (4-1372)] levels. The AUROC value of IL6 [0.66 (0.53-0.79)] was found to be the highest among all followed by IL10 [0.65 (0.51-0.79)], IFNγ [0.63 (0.51-0.77)], PCT [0.61 (0.48-0.75)], and TNFα [0.55 (0.42-0.69)]. Our study suggests that that IL6 is substantially more economical and can reduce the investigation cost to half as compared with the procalcitonin assay.

Elevated serum matrix metalloproteinase-2 levels in heart failure patients with reduced ejection fraction and Cheyne-Stokes respiration.

Cheyne-Stokes respiration (CSR), a kind of central sleep apnea, is referred to as a poor prognostic factor in heart failure patients with reduced ejection fraction (HFrEF). Matrix metalloproteinase (MMP) and B-type natriuretic peptide (BNP) play important roles in HFrEF patients and are markers of poor prognosis. However, there is no literature mentioning the changes in MMP and BNP in HFrEF patients with CSR. From June 2018 to June 2019, 41 adult patients with stable heart failure and left ventricular ejection fraction < 50% were enrolled from the cardiology clinic. After history-taking and medication review to exclude possible central nervous system- or medication-related central sleep apnea, an overnight polysomnography study was performed, and CSR was identified. The morning serum MMP-2, MMP-9, and BNP levels were determined using enzyme-linked immunosorbent assay and fluorescence immunoassay techniques. A positive airway pressure device was applied to 7 patients for 3 months. The serum MMP-2 and BNP levels were significantly higher in HFrEF patients with CSR than in patients without CSR. In addition, elevated serum MMP-2 levels correlated well with the severity of sleep apnea and intermittent hypoxia, which were represented as the apnea-hypopnea index and the oxygen desaturation index. No positive correlation was found between those markers and left ventricular ejection fraction. Finally, the treatment of sleep apnea with continuous positive airway pressure for 3 months tended to reduce the elevated serum MMP-2 levels. Higher serum MMP-2 and BNP levels were found in HFrEF patients with CSR. Elevated MMP-2 levels were correlated with the severity of sleep apnea and intermittent hypoxia. Chuang L-P, Pang J-HS, Lin S-W, etal. Elevated serum matrix metalloproteinase-2 levels in heart failure patients with reduced ejection fraction and Cheyne-Stokes respiration. J Clin Sleep Med. 2022;18(5):1365-1373.

&lt;i&gt;Toxoplasma gondii&lt;/i&gt; Infection in Pregnant Women: Seroprevalence and Associated Risk Factors in Ouagadougou, Burkina Faso

Toxoplasma gondii (T. gondii) infection can lead to abortion in pregnant women. Unfortunately, very limited information is available concerning the seroprevalence and associated risk factors in pregnant women in Burkina Faso. Therefore, the present study aimed to estimate the seroprevalence of T. gondii among pregnant women attending for antenatal care. A cross-sectional study was conducted containing 579 pregnant in Ouagadougou, Burkina Faso. Serological patterns were assessed by enzyme-linked fluorescent immunoassay and the study utilized univariate analysis to identify the potential risk factors for T. gondii infection. Of the 579 pregnant women investigated, 29.71% were tested as T. gondii-seropositive, with 25.91% seropositive for T. gondii IgG antibodies, 1.9% positive for IgM and 1.9% positive for both IgM and IgG. We found firstly that pregnant women consuming unpasteurized cow's milk had a significantly higher seroprevalence than individuals who did not consuming (30.15% vs 06.67%; p=0.0039). Then, age was associated with an increased risk of being seropositive for T. gondii, seroprevalence increases significantly with age, ranging from 18% for 15-25 year olds to 45% for those over 35 years old, pT. gondii (p=0.0065 and 0.023, respectively). The other risk factors like contact with soil, drinking water source, contact with cats and eating some under-cooked meat were not significantly associated with T. gondii infection. Altogether, our study showed that seroprevalence of T. gondii was mainly related to consumption of unpasteurized cow's milk, to patient age, to the number of pregnancies and to living children. The findings will provide key and baseline data for prevention and control of toxoplasmosis among pregnant women and other people.

Predictors of CMV Infection in CMV-Seropositive Kidney Transplant Recipients: Impact of Pretransplant CMV-Specific Humoral Immunity.

BackgroundAlthough cytomegalovirus (CMV)-seropositive solid organ transplant recipients have a relatively lower risk of CMV infection than CMV-seronegative recipients who receive allograft from CMV-seropositive donors, some patients remain at risk of CMV infection after transplant. We investigated the pretransplant CMV-specific humoral immunity (CHI) and other CMV infection predictors in CMV-seropositive kidney transplant (KT) recipients.MethodsThis retrospective study was conducted on adult CMV-seropositive KT recipients during 2017 and 2018. The cumulative incidence of CMV infection was estimated using the Kaplan-Meier method. CHI, measured with an enzyme-linked fluorescent immunoassay and other predictors for CMV infection, was analyzed using Cox proportional hazards models.ResultsOf the 340 CMV-seropositive KT recipients (37% female; mean ± SD age, 43 ± 11 years), 69% received deceased-donor allograft and 64% received induction therapy. During a mean follow-up of 14 months, the cumulative incidence of CMV infection was 14.8%. In multivariate analysis, low pretransplant CHI (defined as anti-CMV immunoglobulin [IgG] titer <20 AU/mL) was significantly associated with CMV infection (hazard ratio [HR], 2.98; 95% CI, 1.31–6.77; P = .009). Other significant predictors of CMV infection included older donor age (HR, 1.03; 95% CI, 1.01–1.06; P = .005), antithymocyte induction therapy (HR, 2.90; 95% CI, 1.09–7.74; P = .033), and prolonged cold ischemic time (HR, 1.06; 95% CI, 1.02–1.10; P = .002).ConclusionsA low pretransplant CHI is independently associated with post-transplant CMV infection in CMV-seropositive KT recipients. A quantitative anti-CMV IgG assay could potentially stratify CMV-seropositive patients at risk of CMV infection after KT.

A study on the role of mid trimester serum beta Human Chorionic Gonadotropin as a predictor of hypertensive disorders of pregnancy

Objective: To determine whether increase in serum beta human chorionic gonadotropin between 12 to 20 weeks of gestation is associated with increase in incidence of gestational hypertension, pre-eclampsia and eclampsia later in pregnancy. Materials and Method: A prospective observational study was undertaken in the department of O&G, SCB Medical College, Cuttack for a period of one year. One hundred pregnant women were enrolled between 12 to 20 weeks of gestation for the study. Their serum beta human chorionic gonadotropin was measured by enzyme linked fluorescence immuno assay and all were followed up. Those who developed hypertension or pre-eclampsia or eclampsia during follow up were included in the hypertensive disorders of pregnancy group and rest were included in normal group. Both the groups were compared and analyzed. Results: Out of the one hundred study samples 14% women developed hypertensive disorders later in pregnancy and the rest 86% remained normotensive. The beta human chorionic gonadotropin levels of the women in the hypertensive group was found to significantly higher than women in normal group. Conclusion: Quantitative estimation of serum beta human chorionic gonadotropin in mid trimester is a very useful screening tool for the prediction of hypertensive disorders of pregnancy. It should be adopted in the routine antenatal care so that there can be a drastic reduction in the maternal mortality and morbidity. Keywords: Hypertensive disorders of pregnancy human chorionic gonadotropin maternal mortality

Development of an Immunofluorescence Assay Module for Determination of the Mycotoxin Zearalenone in Water.

Project Aquafluosense is designed to develop prototypes for a fluorescence-based instrumentation setup for in situ measurements of several characteristic parameters of water quality. In the scope of the project an enzyme-linked fluorescent immunoassay (ELFIA) method has been developed for the detection of several environmental xenobiotics, including mycotoxin zearalenone (ZON). ZON, produced by several plant pathogenic Fusarium species, has recently been identified as an emerging pollutant in surface water, presenting a hazard to aquatic ecosystems. Due to its physico-chemical properties, detection of ZON at low concentrations in surface water is a challenging task. The 96-well microplate-based fluorescence instrument is capable of detecting ZON in the concentration range of 0.09–400 ng/mL. The sensitivity and accuracy of the analytical method has been demonstrated by a comparative assessment with detection by high-performance liquid chromatography and by total internal reflection ellipsometry. The limit of detection of the method, 0.09 ng/mL, falls in the low range compared to the other reported immunoassays, but the main advantage of this ELFIA method is its efficacy in combined in situ applications for determination of various important water quality parameters detectable by induced fluorimerty—e.g., total organic carbon content, algal density or the level of other organic micropollutants detectable by immunofluorimetry. In addition, the immunofluorescence module can readily be expanded to other target analytes if proper antibodies are available for detection.

Evaluation of MicroRNA-122 as a Biomarker for Chronic Hepatitis C Infection and as a Predictor for Treatment Response to Direct-Acting Antivirals.

BackgroundTreatment response to antiviral drugs is a challenging issue in patients with chronic hepatitis C virus (HCV) infection. Although microRNA-122 represents the majority of the microRNA content in hepatic tissues, few studies have evaluated its role in the treatment response, so we aimed to study its role in chronic HCV patients and in predicting the treatment response to direct-acting antivirals (DAAs).MethodsThe study included 125 chronic HCV patients (89 naïve and 36 with a prior failed peginterferon/ribavirin response) and 50 apparently healthy subjects. Complete blood count, liver function, α-fetoprotein, lipid profiles, serum creatinine, abdominal ultrasound, and FibroScan® were assessed. Viral markers, HCV antibodies, and hepatitis B surface antigen were measured by enzyme-linked fluorescent immunoassay, with quantitative estimation of HCV RNA and microRNA-122 levels by real-time PCR.ResultsThe microRNA-122 level in HCV patients (those with a sustained virologic response 12 weeks after finishing therapy [SVR12] and non-responders) was significantly increased compared with controls and expressed more in non-responders versus SVR12 (p=0.042). ROC curve analysis of microRNA-122 for differentiating HCV patients from healthy controls revealed that a cut-off point of >1.45 had a sensitivity of 67.20%, specificity of 94.0%, AUC=0.861, and p<0.001; and for predicting response to treatment a cut-off point ≤5.66 could significantly (p=0.022) predict the occurrence of SVR, with a sensitivity of 60.34%, specificity of 66.67%, and AUC=0.729. Logistic regression analysis showed significant values for microRNA-122 in multivariate and univariate analysis for the prediction of response to DAAs.ConclusionThe results demonstrated the possible function of microRNA-122 as an indicative tool for distinguishing chronic HCV patients from controls and in the assessment of the therapeutic reaction to DAAs.

Differential expression of long noncoding RNA in hepatocellular carcinoma on top of chronic HCV and HBV infections.

Aim of the studyThe task of long noncoding RNAs (lncRNAs) as a prospective goal for hepatocellular carcinoma (HCC) is a candidate for research. Several lncRNAs are involved in signal transduction, directing gene expression and epigenetic alteration in normal and cancer cells. Dysregulation of diverse lncRNAs has been involved in the pathogenesis and progression of different cancers including HCC. We aimed to investigate the differential expression of lncRNAs (aHIF, hPVT1, ANRIL) in HCC on top of chronic hepatitis C virus (HCV) and hepatitis B virus (HBV) infections.Material and methods182 participants were included: 85 patients with HCC in addition to 50 patients with cirrhosis on top of chronic HCV or HBV, and 47 healthy subjects as controls. HCC was diagnosed by triphasic computed tomography (CT). Detection of α-fetoprotein (AFP) and serological markers of HCVAb and HBsAg by enzyme-linked fluorescent immunoassay (ELFA) and quantitation of lncRNAs by real time PCR were applied.ResultsUpregulation of ANRIL and hPVT1 and downregulation of aHIF were observed in patients with HCC on top of HCV and HBV vs. controls. Circulating aHIF could be of major diagnostic importance to discriminate HCC on top of HCV from cirrhotic patients with sensitivity 86.67% and specificity 91.89% whereas circulating hPVT1 had sensitivity 85.0% and specificity 84.62%; moreover ANRIL had AUC 0.902 and could discriminate HCC on top of HBV from cirrhotic patients.ConclusionsThe differential expression of lncRNAs (ANRIL, hPVT1 and aHIF) might be of major worth in predicting the occurrence of HCC in cirrhotic patients related to chronic viral hepatitis and could be beneficial in the early management.

1198. Measurement of Pre-transplant Anti-cytomegalovirus (CMV) Immunoglobulin G Titer to Predict Risk of CMV Infection in CMV-seropositive Kidney Transplant Recipients

BackgroundAlthough cytomegalovirus (CMV)-seropositive solid organ transplant recipients have a lower risk of CMV infection compared with CMV-seronegative recipients, some patients remain at risk of CMV infection after transplant. Low pre-transplant anti-CMV immunoglobulin G (IgG) titer has been reported as a predictor of CMV infection in CMV-seropositive liver and heart transplant recipients, but this association in CMV-seropositive kidney transplant (KT) recipients has not been explored. We investigated the pre-transplant anti-CMV IgG titer and other CMV infection risk factors in CMV-seropositive KT recipients.MethodsThis retrospective study was conducted on CMV-seropositive KT recipients aged >18 years old at Ramathibodi Hospital during 2017 and 2018. The cumulative incidence of CMV infection was estimated with Kaplan–Meier methodology. The pre-transplant anti-CMV IgG titer was measured with an enzyme-linked fluorescent immunoassay. Risk factors for CMV infection were analyzed with Cox proportional hazards models.ResultsOf the 340 included CMV-seropositive KT recipients (37% female; age [mean±SD]: 43±11 years), 69% and 64% received deceased-donor allograft and induction therapy, respectively. The anti-CMV IgG titer was < 20 and >20 AU/ml in 7.1% and 92.9% of patients, respectively. During a mean follow-up of 14 months, the cumulative incidence of CMV infection was 14.8%, including both asymptomatic CMV infection (69%) and tissue-invasive disease (31%). A pre-transplant anti-CMV IgG titer of < 20 AU/ml was significantly associated with CMV infection in both the univariate analysis (HR, 2.70; 95%CI, 1.21–6.05, [p=0.02]) and the multivariate analysis (HR, 2.98; 95% CI, 1.31–6.77, [p=0.009]). Other significant risk factors of CMV infection included older donor age (HR, 1.03; 95% CI, 1.01–1.06, [p=0.005]), anti-thymocyte induction therapy (HR, 2.90; 95% CI 1.09–7.74, [p=0.033]), and prolonged cold ischemic time (HR, 1.06; 95% CI, 1.02–1.10, [p=0.002]).ConclusionA low pre-transplant CMV-specific humoral immunity is independently associated with post-transplant CMV infection in CMV-seropositive KT recipients. The universally available anti-CMV IgG titer test could potentially stratify those at risk and target preventive strategy appropriately.Disclosures All Authors: No reported disclosures

Development of a modified enrichment method for the rapid immunoassay of Escherichia coli O157 strains in fresh cut vegetables

Escherichia coli O157:H7 is a pathogen that colonizes human intestinal epithelial membrane and causes a characteristic lesion and severe syndromes, including diarrhea, hemorrhagic colitis, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura. It is difficult to isolate and identify this pathogen. The purpose of this research was to establish a two-stage enrichment method for the detection of E. coli O157:H7 in fresh cut vegetables. Fresh cut vegetable samples were first enriched in modified EC broth with novo-biocin at 42°C for 6 hr, and then transferred to sorbitol MacConkey broth containing cefixime and tellurite at 35°C for 18 hr. Finally, the enriched broth was verified for the existence of E. coli O157 by using an automated enzyme-linked fluorescence immunoassay (mini-VIDAS) or an immunostrip test. This two-stage enrichment method would decrease the false-positive E. coli O157 tested by immunostrip or mini-VIDAS. The pathogen in food sample could be as low as 10 CFU/g and selectively enriched to 105 or 107 CFU/mL required for the detection by mini-VIDAS or immunostrip. The presence of E. coli O157:H7 in enriched broth was confirmed by multiplex polymerase chain reaction (PCR). Thus, the two-stage enrichment method can be used to rapidly screen food samples for E. coli O157.