Chemiluminescence Immune Assay Research Articles

Hepatitis C virus core antigen: A diagnostic and treatmentmonitoring marker of hepatitis C virus in Indian population.

The diagnosis and treatment monitoring of hepatitis C is quite challenging. The screening test, i.e. antibody assay, is unable to detect acute cases, while the gold standard hepatitis C virus (HCV) reverse transcriptase polymerase chain reaction (RTPCR) assay is not feasible in resource-limited countries such as India due to high cost and infrastructure requirement. European Association for the Study of the Liver and World Health Organization have approved a new marker, i.e. HCV core antigen (HCVcAg) assay, as an alternative to molecular assay. In this study, we have evaluated HCVcAg assay for diagnosis and treatment monitoring follow-up in Indian population infected with hepatitis C. Blood specimen of 90 clinically suspected cases of acute hepatitis C were tested simultaneously for anti-HCV antibody assay via ELISA (enzyme-linked immunoassay), HCVcAg assay by chemiluminescence immune assay (CLIA) and HCV RTPCR VL (viral load) assay. Thirty-four HCV RTPCR positive patients were further enrolled in treatment monitoring group whose blood samples were tested at thebeginning of treatment, twoweeks, fourweeks and 12weeks via HCV core Ag assay and HCV RTPCR Viral Load assay. Considering HCV RTPCR as gold standard, diagnostic performance of HCV core Ag assay and anti-HCV antibody assay was evaluated. The sensitivity and specificity of HCV core Ag assay were higher than that of anti-HCV Antibody assay, i.e. 88.3% and 100% vs. 23.3% and 83.3%, respectively. The overall diagnostic accuracy of HCV core Ag assay was 92.20%. Among treatment follow-up group, HCV core Ag levels correlated well with HCV viral load levels, at the beginning of treatment (baseline) till 12weeks showing highly significant Spearman rank correlation coefficient of > 0.9 with HCV viral load levels. HCV core Ag assay is a cost-effective, practically feasible substitute of HCV RTPCR viral load assay for diagnosis as well as long duration treatment monitoring of hepatitis C infection in resource-limited settings.

SERUM TESTOSTERONE AS A MARKER OF RESPONSE TO ANDROGEN DEPRIVATION THERAPY IN METASTATIC PROSTATE CANCER

Introduction: Prostate cancer (PCa) is the second most widespread cause of cancer and the sixth foremost cause of cancer death among men worldwide. The sustained presence of particular hormones and growth factors stimulates the prostate, such as other sex accessory tissues, in its growth, maintenance, and secretory function. In the current study, we investigated the association of serum testosterone level and prostate-specific antigen (PSA) level before androgen deprivation therapy in metastatic PCa (mPCa) treated with hormone therapy. Methods: This retrospective cohort was conducted from July 2017 to December 2019 in the Department of Urology, Post Graduate Institute of Medical Education and Research, Chandigarh on patients diagnosed with mPCa of age >40 years. Total serum testosterone was measured by Electro chemiluminescent immune assay before the treatment. PSA in serum specimens was evaluated. Results: The mean age was 67.74±8.12 (55–80) years. Serum PSA levels at baseline, 3 months were 530.98±617.28 (ng/dL), and 56.31±89.04 (ng/dL), respectively. The baseline S. testosterone was 288.89±246.53 (ng/mL). Weak negative correlation between the two (p=0.051) As the serum testosterone levels decrease the PSA levels increase and vice-versa. Positive correlation between baseline testosterone and PSA decline (p<0.05). On histopathology, 58.7% (n=71) of the patients had perineural involvement, while 41.3% (n=50) did not have perineural involvement. Conclusion: We conclude that the effect of testosterone might have a possible role in the assessment of treatment response as assessed by PSA. However, the exact implication and its role in disease assessment need to be examined in a larger prospective cohort.

A study of analysis on prevalence, serological marker and prognosis of tuberculosis in tertiary care hospital

BackgroundTuberculosis is an infectious disease responsible for a significant cause of ill health. According to the WHO global tuberculosis report 2021. 9.9 million cases fell sick with TB in 2020. Significantly, the prevalence of tuberculosis in India is 25%. ObjectiveTo analyze the prevalence of tuberculosis in the suburban areas of the metropolitan city in South India. To analyze the serological marker and prognosis of tuberculosis among males and females. To determine the importance of molecular testing - PCR confirmation on TB after AFB smear. MethodsA retrospective study to analyze 462 patients enrolled by the respiratory medicine department on suspecting pulmonary- 356 (M−264 & F-92) and extra-pulmonary-106 (M-73&F-33) patients and diagnosed Zhiel-Neelsen staining, Mantoux test, Chip-based RT-PCR test, Erythrocyte sedimentation rate, and analyzed serological test such as C-Reactive Protein, Chemiluminescence immune assay. Results23 patients were positive in Ziehl-Neelsen staining, 65 were positive in molecular True-Nat PCR test, Mantoux skin test induration in 10 patients, 98 TB Positive patients examined in the serological analysis, 1 & 3 patients reacted in HIV/HBsAg, and HBsAg test respectively, by chemiluminescence immunoassay, 8 PTB and 4 EPTB and 47 non-TB patients were positive in C-reactive protein, 46 TB and 94 non-TB patients detected abnormal values out of these 160 patients in ESR test. ConclusionThe Prevalence of tuberculosis is significantly rising, especially in the middle-aged population. The rapid molecular diagnostics to detect TB are highly sensitive and specific. Serological markers are essential for the analysis of disease prognosis and need to focus on the guidance of DOTS and RNTCP to End TB.

Study of Testosterone Levels in type-II Diabetes Mellitus Patients in North Karnataka Population

Background: Type-II DM is a major health problem globally. There are various complications related to diabetes mellitus, among them low testosterone levels in men is very obvious, health problem which affects the reproduction and sexual life.Method: 250 type-II DM patients at different age groups were studied and compared with 150 controlled groups.The clinical investigation included- FBS, PPBS, Blood urea, S. creatinine, HBA1C, lipid profile, Urine for albumin creatinine ratio, S. testosterone was estimated by using chemiluminescence immune assay and HbA1c by HPLC.Results: BMI in type-II DM group was 25.83 (±2.33) and controlled group was 24.8 (±3.22). Mean HBA1C in type-II DM group was 8.81 (±1.93) and 4.80 (±0.402) in controlled group. S. testosterone level was 119.11 (±86.45) in type-II DM group and 402 (± 171.52) in controlled group.Conclusion: This study revealed that higher sugar level is inversely proportion to lower testosterone levels. It will be a guide to physician, endocrinologist to treat such patients efficiently to elevate the testosterone levels

Preparation of the luciferase-labeled antibody for improving the detection sensitivity of viral antigen

BackgroundViral antigen detection test is the most common method used to detect viruses in the field rapidly. However, due to the low sensitivity, it can only be used as an auxiliary diagnosis method for virus infection. Improving sensitivity is crucial for developing more accurate viral antigen tests. Nano luciferase (Nluc) is a sensitive reporter that has not been used in virus detection.ResultsIn this study, we produced an intracellularly Nluc labeled detection antibody (Nluc-ch2C5) and evaluated its ability to improve the detection sensitivity of respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens. Compared with the traditional horse-radish peroxidase (HRP) labeled antibody (HRP-ch2C5), Nluc-ch2C5 was 41 times more sensitive for inactivated SARS-CoV-2 virus by sandwich chemiluminescence ELISA. Then we applied Nluc-ch2C5 to establish an automatic magnet chemiluminescence immune assay (AMCA) for the SARS-CoV-2 viral spike protein, the limit of detection was 68 pfu/reaction. The clinical sensitivity and specificity reached 75% (24/32) and 100% (48/48) using 32 PCR-positive and 48 PCR-negative swab samples for clinical evaluation, which is more sensitive than the commercial ELSA kit and colloid gold strip kit.ConclusionsHere, monoclonal antibody ch2C5 served as a model antibody and the SARS-CoV-2 served as a model pathogen. The Nluc labeled detecting antibody (Nluc-ch2C5) significantly improved the detection sensitivity of SARS-CoV-2 antigen. This labeling principle applies to other viral infections, so this labeling and test format could be expected to play an important role in detecting other virus antigens.

Antibody Response to SARS-CoV-2 in Relation to the Contributing Factors in COVID-19 Patients.

Human health has always been challenged by variety of viral infections, but severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has surpassed all previous viral diseases and emerged as a major health challenge around the globe. Real-time polymerase chain reaction (PCR) is the gold standard for the diagnosis of SARS-CoV-2 and serological assay provides a compliment to diagnosis after second week of infection. The aim of the study is the characterization of antibody response to SARS-CoV-2 in the blood sample of diagnosed coronavirus disease 2019 (COVID-19) patients, and its potential association with factors such as age, gender, time, and symptoms. Serum from 248 confirmed SARS-CoV-2 patients was investigated for antibodies. Elecsys anti-SARS chemiluminescent immune assay was performed for the detection of nucleocapsid-specific antibodies. Association of antibody response with gender, age, and time after onset of symptoms was analyzed. Among 248 PCR positive SARS-CoV-2 patients, 214 (86.3%) have virus-specific antibody signals. Antibodies positivity rate was higher in male patient patients as compared with female patients (90.8% vs. 79.2%, p = 0.009). Patients aged 30-40 years had the highest antibody positivity rate as compared with other groups (89.10%, p = 0.04). Patients age group >60 years had a lower positivity rate (75%, p = 0.04). The increasing trend in the antibodies detection with time was observed, maximum positive antibodies response rate observed at 8 weeks. Patients were categorized on the basis of clinical symptoms into asymptomatic, mild, and moderate; 17.7% were asymptomatic, 60.5% showed mild symptoms, and 21.8% showed moderate symptoms of the disease. Males were seen to be more asymptomatic as compared with females (i.e., 59.1% to 40.9%). The serological test for SARS-CoV-2 has a high sensitivity at >2 weeks after the positive PCR result or onset of illness. In addition, the serological response differs among patients based on gender, age, as well as time between the onset of symptoms or PCR confirmation and sample collection for the study of antibody response.

A study on periconceptional folic acid supplement intake and serum folic acid levels in pregnant mothers

BackgroundSurvey-based studies have examined the timing of receiving periconceptional folic acid supplementation. To assess the impact of the periconceptional folic acid supplementation, a postulate that multigravida mothers are more likely to have received the supplementation and the level of serum folic acid in them assayed during the first trimester is likely to be higher than primigravida mothers was put forth. Serum folic acid levels were measured in primigravida and multigravida mothers during the first trimester. MethodsOne hundred twenty primigravida and multigravida mothers registered at antenatal clinic of a tertiary care referral centre were included. Serum folic acid assay from samples collected during the first trimester was carried out by chemiluminescence immuneassay. The mothers were followed up during subsequent OPD visits, during admission for delivery and through mobile phones for assessing the delivery outcomes. World Health Organization cutoff values for serum folic acid were used to analyse the results. ResultsNone of the mothers received folic acid supplement before conception. Mean interval from last menstrual period to receiving the first dose of folic acid supplementation was 71.2 days in primigravida and 67.6 days in multigravida mothers. Overall, 21/120 (17.5%) of primigravida mothers and 34/120 (28.3%) of multigravida mothers had serum folic acid values less than 6 ng/ml (deficiency and possible deficiency). ConclusionNone of the mothers received folic acid supplements before conception. Significant proportion of mothers, particularly the multigravida having less than normal levels serum folic acid indicates correctable lacunae amenable for preventive intervention.

Correlation between vitamin D deficiency and fasting blood glucose levels in obese children.

This study aimed to determine the relationship between Deficiency of vitamin D and hyperglycemia in children with obesity. This cross-sectional study was carried out between February and April of 2020. This study's subject population consisted of high school students aged 11 to 17 who were obese. The 25 (OH) level of Vitamin D was determined using the Chemiluminescence Immune Assay method. Subjects were divided into 4 groups; vitamin D deficiency with hyperglycemia, vitamin D deficiency without hyperglycemia, normal vitamin D with hyperglycemia, and normal vitamin D without hyperglycemia. The prevalence of hyperglycemia was 28 (54.9%) in the vitamin D deficiency group and 17 (378%) in the normal vitamin D group, respectively. Statistical analysis revealed no statistically significant difference between the two groups (p=0.093). With p=0.031, there was a significant difference in the mean value of fasting blood sugar levels between the two groups. The frequency of hyperglycemia was 54.9 percent in the vitamin D deficiency group and 37.8 percent in the normal vitamin D group. Fasting blood sugar levels were lower in obese children with vitamin D deficiency than in children with normal vitamin D. Fasting blood sugar levels were correlated with vitamin D levels in obese children, but clinically insignificant. However, the research is ongoing, so, it is critical to conduct early screening for Vitamin D and fasting blood sugar levels in order to reduce the risk of metabolic and cardiovascular diseases.

Correlation Between Serum Ferritin and Gonadotrophins and Sex Hormones in Patients with Transfusion Dependent â-Thalassemia

Background: Patients with transfusion dependent beta thalassemia major develop a number of complications of whichhypogonadism is the most common. In addition, raised serum ferritin level has been associated with hypogonadism.Objective: To determine a correlation between serum ferritin, gonadotrophins and sex hormones in patients with transfusiondependent â-thalassemia syndrome patients.Material and methods: This cross sectional study was designed and conducted at the Institute of Basic Medical Sciences (IBMS),Khyber Medical University (KMU) Peshawar and Fatimid Foundation, blood bank and hematological services Peshawar. Betathalassemia syndrome patients enrolled in the study were 97. Blood samples collected from the study subjects were analyzed atIBMS KMU using chemiluminescence immune assay for determination of serum ferritin and gonadal hormones such as FSH, LH,Estradiol and Testosterone.Results: The mean±SD age of patients was 18.93±3.6 years with a range of 15-32 years. There were 55 (56.7%) males and 42(43.3%) females. Overall hypogonadism was diagnosed in 39 (40.21%) patients. Although we did not find any association betweenserum ferritin level and gonadal function in our study, hypogonadism having males had a higher serum ferritin level (p = 0.02). Astatistically significant negative correlation was found between serum ferritin and serum level of LH, FSH and testosterone in males(p <0.05).Conclusion: The effect of iron overload on serum level of gonadotrophins and gonadal hormones is more pronounced in males ascompared to female thalassemic patients.Keywords: Beta Thalassemia syndrome, Chelation, Ferritin, Hypogonadism, Gonads, Sex Hormones

Dopamine agonists as genital endometriosis target therapy

Objective The present study was to find a pathogenic evidence for dopamine agonist application in patients with endometriosis associated pain syndrome. Patients and technique The study involved 227 patients of reproductive age with histologically confirmed genital endometriosis (GE) of I–III degree according to ASRM classification. The control group included 12 women with no laparoscope detected gynecologic pathology. The levels of prolactin (PRL), peripheral blood (PB), and peritoneal fluid (PF) were evaluated by chemiluminescence immune assay. The pain syndrome was measured by McGill visual analogue scale. Statistica10 program (StatSoft, Inc., Tulsa, OK) was applied for obtained data processing. Results A correlation was established between GE rate and levels of PRL and PB (Rs = 0.28, p < .05) as well as a correlation of PRL in PB and PF (Rs = 0.29, p < .05). Patients receiving cabergoline combined with hormone therapy standard schemes manifested considerable pain syndrome relief. Conclusions PRL involvement in GE pathogenesis and more intense therapeutic impact on pain syndrome in case of combined administration of dopamine and standard hormone therapy prove cabergoline application in clinical practice.

Impact of Mushroom Pleurotus tuber-regium (Rumph. ex) Fr. Extract on Lipid Profile and Testosterone of Rat

Pleurotus tuber-regium has been used as fodder and traditionally used as medicinal supplement. The aim of this study was to screen in vitro proximate biochemical composition and nutritional potentiality of P. tuber-regium extract for further in vivo analysis of impact of the extract on serum lipid and testosterone profile of rats. Total cholesterol, HDL cholesterol, LDL cholesterol were measured photometrically. Testosterone was measured by chemiluminescence immune assay. P. tuber-regium extract contains different biochemicals such as proteins, carbohydrates, tannins, flavonoids, phenols etc. and showed highly quantified calorific value (297.89±1.92 calori/100g). Acute toxicity test showed no mortality and toxic behavioural symptoms. Significant decrease in total cholesterol (75.52±0.39mg/dL), LDL cholesterol (34.58±1.69mg/dL) and triglyceride (81.31±1.25) were observed in rat group treated with 200mg/kg dose of extract. 400mg/kg dose of extract showed higher efficacy than 200mg/kg dose of extract and significantly decreased total cholesterol (72.25±1.11mg/dL), LDL cholesterol (26.37±1.21mg/dL) and triglyceride (69.42±0.72). The extract showed testosterone enhance efficacy. 400mg/kg dose of extract showed high level testosterone (178.96±0.68 ng/dL) enhance efficacy than 200mg/kg dose (174.64±0.64ng/dL). Due to hypocholesterolaemia and testosterone elevating properties of P. tuber-regium extract, it can be said that, mushroom P. tuber-regium can be consumed as healthy diet supplement to maintain cholesterol level and to maintain good male reproductive health.

Comparison of the 25(OH)D Levels Between Sarcopenia and Frailty in Elder Women: A Cross-Sectional Observation Analytic Study in Elderly Community in Surabaya

The prevalence of vitamin D deficiency in the elderly, specifically in post-menopausal women, are very high. Deficiency of 25(OH)D has a direct impact on decreasing muscle strength and mass that can lead into sarcopenia and frailty. This study aimed to compare the difference levels of 25(OH)D between sarcopenia and frailty in elderly community. Methods: This was a cross-sectional design study. The blood sample were collected to evaluate the levels of 25(OH)D using The Chemiluminescent Immune Assay (CLIA) method. Results: Most of the subjects were at deficiency of 25(OH)D with mean of 16.1 (range level of 6.2-32.7). Subjects with deficiency level of 25(OH)D were 20 (71.4%) with sarcopenia and 23 (85.1%) with frailty. Subjects with an insufficiency level of 25(OH)D were 6 (21.4%) with sarcopenia and 23 (85.1%) with frailty. Subjects with sufficiency level of 25(OH)D were 2 (7.1%) with sarcopenia and 1 (3.7%) with frailty. The median 25(OH)D levels were 17.1 and 13.2 for sarcopenia and frailty, respectively. There was significant differences in levels of 25(OH)D in both groups (p = 0.014). Conclusions: The levels of 25(OH)D of subjects with sarcopenia was significantly higher than those with frailty.

Performance Evaluation of the CLIA and ECLIA for Anti-TP Screening in Blood Donors

To evaluate the performance of the chemiluminescence immune assay (CLIA) and the electro-chemiluminescence immuneoassay(ECLIA) for Treponemapallidum antibody（anti-TP） screening in blood donors. The sero-panel samples from NCCL were tested with ELISA, CLIA and ECLIA assays synchronously to evaluate their performances respectively. The sensitivity and the negative predictive value of the CLIA were 100%, which were the same as one kind of ELISA, and better than the other ELISA; The specificity of the CLIA was 88.46%, the accuracy rate was 97.02%, the positive predictive value was 96.13%, which were higher than both ELISA. Due to the significant interference of sample heat inactivation in ECLIA detection, the result can not demonstrate the true performance of ECLIA in this study. The preliminary result was as follows: the sensitivity was 98.93%, the negative predictive value was 96.75%, and the accuracy rate, specificity and positive predictive value of ECLIA were 97.02%, 91.54% and 97.10% respectively. Compared with ELISA, the CLIA has higher sensitivity and specificity and can be used for Treponemal antibody screening in blood bank. Unfortunately, the data in this study cannot come to a conclusion for ECLIA and needs more testing.

High efficiency of the aldosterone-to-renin ratio in precisely detecting primary aldosteronism.

The aldosterone-to-renin ratio (ARR) is extensively used for primary aldosteronism detection. Chemiluminescence immune assay (CLIA) is newly applied in aldosterone and renin detection for calculating the aldosterone-to-renin ratio. The performance of new ARR in aldosteronism detection is poorly evaluated. We aim to estimate the diagnostic value of this new aldosterone-to-renin ratio by highly standardized and clinically based protocol. Four hundred and forty-two patients were enrolled in our retrospective study. They went to the first affiliated hospital of Sun Yat-sen University with difficult-to-control hypertension. Primary aldosteronism diagnosis was based on clinical criteria, including a saline infusion test and other necessary inspections. ARR was calculated from plasma aldosterone and renin measured by CLIA. The cutoff value was determined and the diagnostic value was evaluated. The cutoff value of ARR for primary aldosteronism diagnosis was 28.3, with a sensitivity of 87.6%, specificity of 100%, negative-predictive value of 96.4%, and positive-predictive value of 100%. Then, we found that Age was weakly correlated with ARR. The cutoff values of ARR for primary aldosteronism diagnosis in 26-45-, 46-65-, and 66-85-year-old patients were, respectively, 29.45, 27.95, and 28.4, with sensitivities of 87.5%, 87.7%, and 87.5%, specificities of 100% for all, negative-predictive values of 97.7%, 94.3%, and 96.3%, and positive-predictive values of 100% for all. ARR generated by CLIA is a good diagnostic test for primary aldosteronism without making a false-positive diagnosis. Although ARR is correlated with age, ARR cutoff values for different ages are not more efficient than that for total sample in primary aldosteronism diagnosis.

Effects of Short Term Alendronate Administration on Bone Mineral Density in Patients with Chronic Kidney Disease.

Abstract Background: Osteoporosis is highly prevalent in CKD patients and is characterized by low bone mass leading to decreased bone strength. It is associated with an increased risk of fracture, thus increasing morbidity and mortality. Bisphosphonate administration decreases fracture risk in postmenopausal females with osteoporosis. There are limited studies showing effects of short term alendronate administration on BMD in predialysis osteoporotic patients with CKD. Methods: This study was conducted on fifty adult patients with chronic kidney disease. Patients were divided into two groups. Group A consisted of seventeen patients with CKD stage 3 (eGFR 45-30 ml/min/1.73m2) and Group B comprised thirty three patients with CKD stage 4 (eGFR 30-15 ml/min/1.73m2). The study included male patients between age 18-75 years and premenopausal non pregnant females older than 18 years of age. All the patients were osteoporotic having T score &lt; −2.5 on DEXA scan. The patients were administered 70 mg alendronate tablet once a week for 6 weeks. Renal parameters, parathyroid hormone, calcium, phosphorous and alkaline phosphatase levels were assayed at baseline for 6 months. Serum (iPTH) level (pg/ml) was measured by chemiluminescent immune assay (CLIA) method and serum 25 Hydroxy Vitamin D level (ng/ml) was measured by enzyme linked immunosorbent assay (ELISA) method. Bone Mineral Density (BMD) was measured at baseline for 6 months, by dual energy x-ray absorptiometry at lumbar spine and neck of femur and lowest values were included. The results were obtained for T score, Z score and bone mineral density (g/cm2). Results: The BMD, T score and Z score increased in both groups after 6 months with a statistically significant difference in the treatment group. In Group A, T score, Z score and BMD (g/cm2) increased from −2.60±0.086, −2.13±0.28, and 0.80±0.008 at baseline to −2.57±0.097, −2.11±0.26 and 0.81±0.008 after six months. In Group B, the T score, Z score and BMD (g/cm2) increased from −3.17±0.24, −2.82±0.33 and 0.738±0.03 to −3.16±0.25, −2.66±0.95 and 0.743±0.03 after six months with a statistically significant difference. eGFR decreased in both groups but the difference was statistically non-significant (P&gt;0.05). The serum iPTH levels increased after 6 months in both groups with a statistically insignificant difference. There was an increase in serum calcium and decrease in serum phosphate levels after six months, however the difference was statistically insignificant (p&gt;0.05). The SAP values decreased in both groups after six months with a statistically insignificant difference. The main side-effect in the alendronate group was the occurrence of gastroesophageal reflux symptoms in two subjects. Conclusion: Low-dose alendronate, administered for a limited duration, appears to be well tolerated in CKD patients. The BMD increased in both groups suggesting a bone-preserving effect of alendronate.

Cardiac troponin T as a marker of myocardial injury in a group of asphyxiated African neonates

Background: In caucasian newborns, troponin T (cTnT) is a specific marker for myocardial injury in perinatal asphyxia. This is the first such study in negroid neonates.Aims: To evaluate myocardial injury in a group of asphyxiated African newborns and determine the influence of mode of delivery on cTnT levels.Methods: Serum cTnT and clinical parameters in 40 asphyxiated and 40 healthy negroid neonatal controls were measured within the first 72 hours of life by chemiluminescence immune-assay. Perinatal asphyxia was assessed by APGAR score. The infants were followed up until discharge or death.Results: Mean (SD) cTnT values were significantly higher in asphyxiated infants [0·03 (0·04) ng/ml] than in healthy controls [0·01 (0·006) ng/ml, P = 0·002]. Asphyxiated infants delivered by forceps and vacuum extraction had significantly higher levels of cTnT [0·04 (0·018) ng/ml] than those born by caesarian section [0·02 (0·008) ng/ml] and by normal delivery [0·03 (0·01) ng/ml, P = 0·003]. cTnT levels were higher in infants who died (P = 0·037).Conclusion: In the asphyxiated infants, mean cTnT levels were significantly higher than in controls. They were also significantly higher in those born by vacuum and forceps delivery and asphyxiated infants who died.

Correlation of free fetal DNA with beta-human chorionin gonadotropin in circulation in pregnant women with high risk of Down's syndrome

To investigate significance and correlation of free fetal DNA (fDNA) and beta-human chorionic gonadotropin (beta-hCG) in circulation in pregnant women with high-risk of Down's syndrome (DS). Pregnant women with a male fetus at second trimester screening for Down's syndrome were chosen, including 5 women with a trisomy 21 fetus (DS group), 21 women with DS high-risk pregnant women (DS high-risk group) matched with 22 normal pregnant women as control group. Free fDNA in maternal plasma were extracted. Male DYS14 gene was labled as fDNA, real-time PCR was used to detect fDNA expression. The concentration of beta-hCG in maternal serum was detected by chemiluminescence immune assay. The relationship between level of free fDNA and beta-hCG concentration was analyzed by Pearson correlation analysis. (1) The mean level of free fDNA was (127 +/- 58) GE/ml in DS group, which was significantly higher than (78 +/- 28) GE/ml in DS high-risk group and (48 +/- 21) GE/ml in control group,respectively (P < 0.01). When compared the level of free fDNA between DS high-risk group and control group, it reached statistical difference (P < 0.01). (2) The mean concentration of beta-hCG was (97 +/- 43) kU/L in DS group, which was significantly higher than (58 +/- 25) kU/L in DS high-risk group and (38 +/- 19) kU/L in control group, respectively (P < 0.01). The level of beta-hCG in DS high-risk group was also significantly higher than control group (P < 0.01). (3) The positive relationship between the level of free fDNA in maternal plasma and beta-hCG concentration in maternal serum was observed among three groups (r = 0.83, P < 0.05; r = 0.76,P < 0.01; r = 0.86,P < 0.01). Free fDNA in maternal plasma might be a candidate marker used for prenatal DS screening. However, its clinical value need to be evaluated because of positive correlation between free fDNA and beta-HCG in maternal circulation.

Individuazione di un nuovo marker da impiegare per una corretta stadiazione dell’infezione da EBV

Epstein Barr Virus (EBV), also classified as Human Herpes Virus 4, infects the vast majority of adults worldwide and establishes both non-productive (latent) and productive (lytic) infection. Classical EBV diagnosis includes quantitative determination of viral DNA and serological analysis, based on the determination of IgG and IgM responses against the viral capsid antigen (VCA) and the IgG response against the EBV nuclear antigen-1 (EBNA-1). EBV-serology can be misleading in some cases, such as acute infections with low or undetectable VCA IgM, convalescent patients with persistent or reactivated VCA IgM and negative anti- EBNA-1 IgG, due to a loss of this marker during immunosuppression. In all these cases avidity determination of IgG is helpful to prevent false diagnosis. Avidity represents the stability of the antigen-antibody interaction. Its value increases during the infection, so high avidity never associates with a primary acute infection. We studied the importance of avidity determination of p18 (VCA)-IgG to achieve unequivocal interpretation of serological results. The amount of IgG and IgM is determined by Chemiluminescent Immune Assay (CLIA), a rapid and highly sensitive method suitable for automation. The intensity of luminous signal produced by antibody-antigen recognition is expressed as Relative Light Unit (RLU). p-18 IgG is determined using a recombinant p18 antigen expressed in E. coli. Avidity index is determined in CLIA by the ratio between denaturated and not denaturated IgG specific antibodies expressed in RLU. These results demonstrate that avidity determination represents an important additional marker particularly in cases with aberrant serological profile.