Showing papers in "Journal of Clinical Laboratory Analysis in 2011"
TL;DR: Serum indoxyl sulfate level was closely associated with p‐cresylsulfate level in CKD patients and there was a positive correlation between indoxy sulfate and p‐ cresylSulfatelevel.
Abstract: Indoxyl sulfate and p-cresylsulfate was associated with poor clinical outcome of uremia. We explored the relationship between the two toxins and renal function in chronic kidney disease (CKD) patients. This study enrolled 103 stable CKD patients (stage 3–5 and hemodialysis (HD) patients). Serum levels of indoxyl sulfate and p-cresylsulfate were measured using ultra performance liquid chromatography. General laboratory results and patient background were also checked. Patients with advanced CKD had higher serum indoxyl sulfate, p-cresylsulfate based on ANOVA test. There were significant correlation between indoxyl sulfate and p-cresylsulfate and serum creatinine after multivariate regression analysis (B=3.59, P<0.01; B=0.93, P=0.04, respectively). In addition, there was a positive correlation between indoxyl sulfate and p-cresylsulfate level (r=0.61, P<0.01). Indoxyl sulfate and p-cresylsulfate level increased gradually while renal function declined and reached the peak at the stage of HD. Serum indoxyl sulfate level was closely associated with p-cresylsulfate level in CKD patients. J. Clin. Lab. Anal. 25:191–197, 2011. © 2011 Wiley-Liss, Inc.
108 citations
TL;DR: It is suggested that MPV might help in the assessment of fibrosis in patients with chronic hepatitis B and should not be considered a stand‐alone test for this use owing to nonspecificity with other diseases.
Abstract: Introduction: Many noninvasive tests have been studied for the diagnosis and determining the liver fibrosis score (LFS). In this study, we aimed to research the correlation of mean platelet volume (MPV) and stage of liver fibrosis in patients with chronic hepatitis B (CHB). Patients and Methods: Fifty-nine patients with CHB were enrolled retrospectively into the study. Age–sex matched 25 healthy subjects were used as control group. The following data were obtained from computerized patient registry database: HBV-DNA level, hepatitis B e-antigen seropositivity, liver enzymes and function tests, white blood cell count, platelet count, hemoglobin, histological activity index, LFS, and MPV. Patients were divided into two groups: patients without significant fibrosis (F0, F1, or F2) (Group 1) and patients with advanced fibrosis (F3, F4) (Group 2). Results: A statistically significant increase in MPV was seen in patients with CHB compared with healthy controls (8.49±0.84 fl vs.7.65±0.42 fl, P<0.001). Receiver operating characteristic curve analysis suggested that the optimum MPV level cut-off points for CHB was 8.0 fl, with sensitivity, specificity, PPV, and NPV of 68, 76, 86, and 50%, respectively. MPV levels were significantly higher in Group 2 (8.91±0.94 fl, P: 0.009) compared with Group 1 (8.32±0.74 fl). ROC curve analysis suggested that the optimum MPV level cut-off points for Group 2 was 8.45 fl, with sensitivity, specificity, positive and negative predictive value of 77, 59, 45, and 85%, respectively. Multivariable logistic regression model, which consisted of HAI, ALT, HBV-DNA, platelet count, and MPV, was performed. We showed that MPV was independently associated with advanced fibrosis (P: 0.031). Conclusion: We suggest that MPV might help in the assessment of fibrosis in CHB. It should not be considered a stand-alone test for this use owing to nonspecificity with other diseases. J. Clin. Lab. Anal. 25:162–165, 2011. © 2011 Wiley-Liss, Inc.
76 citations
TL;DR: Serum GPC3 levels were significantly higher in patients with HCC and cirrhosis compared with control subjects and there was no correlation between G PC3 levels and prognostic parameters.
Abstract: α-Feto protein (AFP) is the widely used tumor marker in the diagnosis of hepatocellular carcinoma (HCC). The aim of this study was to assess the diagnostic and prognostic validity of a novel marker, serum Glypican-3 (GPC3) and to compare AFP in patients with HCC. One hundred and twenty-eight patients (75 patients with HCC, 55 patients with cirrhosis, and 28 healthy controls) were included in this study. Cut-off value of GPC3 was 3.9 pg/ml. AFP was divided into four subgroups, according to cut-off values with 13, 20, 100, and 200 ng/ml. Sensitivity, specificity, and positive and negative predictive values of GPC3 and AFP13, AFP20, AFP100, AFP200 subgroups and also GPC3+AFP13, GPC3+AFP20 , GPC3+AFP100 , GPC3+AFP200 combinations were compared. Serum GPC3 levels were significantly higher in patients with HCC and cirrhosis compared with control subjects (P<0.05). The median serum GPC3 levels were 3.9 pg/ml in controls, 5.51 pg/ml in patients with cirrhosis, and 5.13 pg/ml in those with HCC. The median serum AFP levels were 1.37 ng/ml in controls, 2.32 ng/ml in cirrhotics, and 50.65 ng/ml in HCC patients. The sensitivity, specificity, and positive and negative predictive values of GPC3 was 61.33, 41.82, 58.97, and 44.43%, respectively. The values for AFP were 68.57, 94.55, 94.12, and 70.27%, respectively. There was no correlation between GPC3 levels and prognostic parameters. GPC3 is not a useful diagnostic and prognostic marker for HCC.
49 citations
TL;DR: It is suggested that the PCR‐RFLP assay with serosity materials punctured from CL patients using Hae III enzyme is useful for the rapid identification of Leishmania species.
Abstract: This study was aimed at identifying the Leishmania species using serosity materials punctured from skin lesions of cutaneous leishmaniasis (CL) patients by using internal transcribed spacer1 (ITS1) polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). We used the PCR-RFLP on 60 parasitological confirmed CL patients who referred to leishmaniasis laboratory from the School of Public Health, Tehran University of Medical Sciences. The PCR-RFLP could correctly detect 51 Leishmania species of the 60confirmed positive specimens, where all the other 10 parasitological (microscopy and culture) negative samples that were prepared from other bacterial- and fungal-infected lesions had negative results. The results also revealed that Leishmania major was the dominant species (53.3%). This study suggests that the PCR-RFLP assay with serosity materials punctured from CL patients using Hae III enzyme is useful for the rapid identification of Leishmania species. J. Clin. Lab. Anal. 25:20–24, 2011. © 2011 Wiley-Liss, Inc.
48 citations
TL;DR: An increased serum Hcy may be a precipitating factor for vitiligo in the predisposed individuals, and its level was high in male patients as compared with female patients.
Abstract: Background: Vitiligo is an acquired depigmenting disorder caused by the destruction of melanocytes. The exact etiopathogenesis and mechanisms of vitiligo are not fully understood. Vitamin B12 and folic acid levels are decreased in vitiligo, which are the important cofactors required in the metabolism of Homocysteine (Hcy). Consequently, Hcy level increases in the circulation. Therefore, it is possible that increased Hcy plays a role in melanocytes destruction. The aim was to study for any association of vitiligo with serum Hcy level. Method: A total of 30 vitiligo patients of both sexes and 30 control subjects were enrolled in this study. Sera from patients and controls were assayed for Hcy by Enzyme immunoassay. The collected data were analyzed by SPSS version-16. Results: The mean serum level of Hcy was significantly higher in patients with vitiligo as compared with healthy controls and its level was high in male patients as compared with female patients. The Hcy level in vegetarian patients was significantly higher as compared with nonvegetarian patients. The Hcy level was also significantly higher in active vitiligo patients as compared with stable vitiligo patients. Conclusion: An increased serum Hcy may be a precipitating factor for vitiligo in the predisposed individuals. Serum Hcy is related to the gender of patients, activity of disease, and dietary habits of vitiligo patients. J. Clin. Lab. Anal. 25:110–112, 2011. © 2011 Wiley-Liss, Inc.
34 citations
TL;DR: This study has objectively shown that the analytical precision of current instruments is being achieved contrary to the known problems surrounding the analytical bias for 25D assays.
Abstract: Background: Measurement of 25-hydroxyvitamin D, (25D) is central in the investigation of pathologies of bone and mineral ion metabolism and in determining a patient's vitamin D status More recently much research interest has lead to investigating the role it can play in decreasing the risk of many chronic illnesses, including common cancers, autoimmune diseases, infectious diseases, and cardiovascular disease Knowledge of the biological variation of an analyte forms an essential part of evaluating a new analyte enabling the objective assessment of the changes in serial results, the utility of reference intervals as well as establishing laboratory quality specifications Methods: This study determined the biological variation of 25D in 20 healthy individuals that was calculated according to the familiar methods outlined by Fraser and Harris Results: The within-subject variation was 121% and the between subject variation was 403% The critical difference for sequential values significant at P<005 was calculated as 384% The within-subject variation forms a relatively small part of the reference interval shown by the low index of individuality of 03 Objective analytical quality goals have also been established which have shown achievable minimum performance for imprecision of ∼6% The desirable analytical bias goal was ∼10% Conclusion: This study has objectively shown that the analytical precision of current instruments is being achieved contrary to the known problems surrounding the analytical bias for 25D assays The limitations of using reference intervals for 25D, both in diagnoses and monitoring are shown J Clin Lab Anal 25:130–133, 2011 © 2011 Wiley-Liss, Inc
30 citations
TL;DR: The results suggest that oxidative–antioxidative balance and collagen turnover are altered by the development of COPD in human lungs, and prolidase activity may reflect disturbances of collagen metabolism in this pulmonary disease.
Abstract: Background: Chronic obstructive pulmonary disease (COPD) is a consequence of an underlying chronic inflammatory disorder of the airways that is usually progressive and causes dysregulation in the metabolism of collagen. Prolidase has an important role in the recycling of proline for collagen synthesis and cell growth. Objective: We measured and compared prolidase activity in healthy individuals with COPD patients to find out that whether its activity might reflect disturbances of collagen metabolism in the patients. We also investigated oxidative–antioxidative status and its relationship with prolidase activity in this disease. Methods: Thirty voluntary patients with COPD and 30 healthy control subjects with similar age range and sex were included into the study. Plasma prolidase activities, total antioxidant capacity (TAC) and lipid peroxidation (LPO) levels were measured in the patient and control groups. Results: Plasma prolidase activity and TAC levels were significantly lower, and LPO levels were significantly higher in the patients than those in the control subjects (P<0.05, P<0.001, and P<0.001, respectively). Significant correlations were detected between plasma prolidase activity and TAC and LPO levels in the patients group (r=0.679, P<0.001; r=−426, P<0.05, respectively). Conclusions: The results suggest that oxidative–antioxidative balance and collagen turnover are altered by the development of COPD in human lungs, and prolidase activity may reflect disturbances of collagen metabolism in this pulmonary disease. Monitoring of plasma prolidase activity and oxidative–antioxidative balance may be useful in evaluating fibrotic processes and oxidative damage in the chronic inflammatory lung disease in human. J. Clin. Lab. Anal. 25:8–13, 2011. © 2011 Wiley-Liss, Inc.
28 citations
TL;DR: The results suggest that according to the criteria of the IDF, the significant decrease observed in serum paraoxonase activity was independent of the metabolic syndrome in patients with mild‐to‐moderate plaque‐type psoriasis, whereas the significant Decrease in serum arylesterase activities was associated with the metabolic Syndrome.
Abstract: Objective: Psoriasis is a chronic immune-mediated inflammatory skin disease associated with metabolic syndrome, which is made up of a cluster of disorders, including obesity, diabetes mellitus, dyslipidemia, and cardiovascular disease. The aim of this study was to investigate serum paraoxonase-1 activities and oxidative status parameters in patients with plaque-type psoriasis with or without metabolic syndrome. Methods: In this study, patients with plaque-type psoriasis with (n=25) or without (n=27) metabolic syndrome, according to the criteria of the International Diabetes Federation (IDF), were matched for age and sex to an equally sized control group (n=25). Results: In patients without metabolic syndrome, serum paraoxonase and arylesterase activities showed mean decreases of 29 and 6%, respectively, whereas in patients with metabolic syndrome, the mean decreases in the enzymes' activities were 35 and 11%, respectively, compared with those in the control group. Serum total antioxidant capacity and total oxidant status were not statistically significant in any of the three groups. Multiple linear regression analysis revealed that HDL cholesterol and log-transformed triglyceride were independent variables for serum arylesterase activity and that fasting glucose and diastolic blood pressure were independent variables for serum paraoxonase activity. Conclusions: These results suggest that according to the criteria of the IDF, the significant decrease observed in serum paraoxonase activity was independent of the metabolic syndrome in patients with mild-to-moderate plaque-type psoriasis, whereas the significant decrease in serum arylesterase activity was associated with the metabolic syndrome. J. Clin. Lab. Anal. 25:289–295, 2011. © 2011 Wiley-Liss, Inc.
27 citations
TL;DR: It was determined that S. aureus treated with agarose, methanol, and lysozyme could be detected with FISH and the 1 hr protocol is a useful alternative to conventional FISH.
Abstract: To detect with whole-cell fluorescence in situ hybridization (FISH), Staphylococcus aureus is typically permeabilized with lyozyme and lysostaphin. We tested whether it was feasible to detect S. aureus and differentiate it from Staphylococcus epidermidis with lysozyme-only permeabilization. We compared lysozyme permeabilizationto S. aureus permeabilized with lysozyme in combination with lysostaphin. It was determined that S. aureus treated with agarose, methanol, and lysozyme could be detected with FISH. The 1 hr protocol is a useful alternative to conventional FISH. J. Clin. Lab. Anal. 25:142–147, 2011. © 2011 Wiley-Liss, Inc.
25 citations
TL;DR: A flexible bead‐based immunoassay to measure total PSA (tPSA) and fPSA simultaneously and is rapid, sensitive, and less expensive, which allows both single sample and high‐throughput measurement of tPSA and f PSA over a wide range of concentrations.
Abstract: Objective: Prostate-specific antigen (PSA) is the most important biochemical tumor marker for the early detection of prostate cancer; however, its diagnostic specificity is low. Therefore, free PSA (fPSA) test is recommended as an adjunct to increase the specificity. However, all the current technology only allows detecting one biomarker at one time. In this study, we reported a flexible bead-based immunoassay to measure total PSA (tPSA) and fPSA simultaneously. Materials and Methods: We used the Luminex xMAP bead array technology to measure tPSA and fPSA at one time, employing two mouse monoclonal anti-PSA antibodies (5G6 and 8A6) for coating and another mouse monoclonal anti-PSA antibody (5A6) for detection. Then wecompared the data of Luminex assay with that of the conventional enzyme-linked immunosorbent assay (ELISA). Results: The assay was fast with a wide dynamic range. The lower detection limit for tPSA and fPSA were 2.3 and 1.3 pg/ml. The inter-assay coefficients for tPSA and fPSA were between 5.64 and 7.65%, and the intra-assay coefficients for tPSA and fPSA were between 4.15 and 5.89%. A close correlation between the new assay and the conventional ELISA was observed. Conclusions: The bead-based platform is rapid, sensitive, and less expensive, which allows both single sample and high-throughput measurement of tPSA and fPSA over a wide range of concentrations. J. Clin. Lab. Anal. 25:37–42, 2011. © 2011 Wiley-Liss, Inc.
24 citations
TL;DR: A significant promise was shown for the use of whole saliva and oral fluid together with the modified commercial EIA for Hepatitis B virus infection surveillance and there was excellent reproducibility.
Abstract: In this study, a modified enzyme immunoassay (EIA) was evaluated for the Hepatitis B surface antigen (HBsAg) among whole saliva and oral fluid samples. Specimens were collected from 115 individuals who gave serum and oral fluid using Salivette (Sarstedt, Numbrecht, Germany) and whole saliva. Saliva specimens were tested following a modified ELISA, and the results were compared with paired serum specimens that were tested according to the supplier's instructions. Transport buffer for the oral fluids, sample volume for assay, incubation period of sample with conjugate as well as cut-off values were evaluated to optimize the assay. The highest sensitivity and specificity were obtained by increasing the incubation of sample and conjugate to 16 hr and using the area under the receiver operating characteristic curve to calculate cut-off values. HBsAg was detected in 40 oral fluids and 44 whole saliva samples out of 47 paired positive serum specimens and not detected in 64 oral fluids and 63 whole saliva samples out of 68 matched negative sera samples by the ELISA assay. There was excellent agreement between the results for the serum and saliva specimens kappa value (κ): 0.80 for oral fluid and κ: 0.87 for whole saliva and there was excellent reproducibility. Using an optimized protocol, the sensitivities of whole saliva and oral fluid were 93.6 and 85.1%, respectively, whereas specificities of whole saliva and oral fluid were 92.6 and 94.1%, respectively. Our data showed a significant promise for the use of whole saliva and oral fluid together with the modified commercial EIA for Hepatitis B virus infection surveillance.
TL;DR: The microscopic cell counter with microchip performed well with high precision, linearity, and efficient running time than both the manual trypan blue and the flow cytometry methods.
Abstract: The cell viability test is an essential tool in any laboratory, performing cell-based studies and clinical laboratory tests. The trypan blue exclusion method is the most popular assay for its simple concept among various diagnostic tools. However, several disadvantages include time-consuming and labor-intensive steps with low precision. In this study, we evaluated a new technique for the automatic cell viability measurement with microscopic cell counter and microchip. Upon blood draw from 11 healthy volunteers, Mononuclear cells were separated immediately from the heparinized whole blood, and the viable cells were diluted from 100 to 1%. The cell viability tests were performed simultaneously with following three methods: the conventional manual trypan blue exclusion method; the flow cytometry measurement with propidium iodide stain; and the newly developed microscopic cell counter with microchip. Linearities, precisions, and correlations from three methods were analyzed and compared. The correlations data from the microscopic cell counter were in good agreement with both the conventional trypan blue method (r=0.99, P<0.05) and the flow cytometry (r=0.99, P<0.05), respectively. The precision (2.0-6.2%) and linearity from the microscopic cell counter method with microchip were superior in comparison with the conventional method. The microscopic cell counter with microchip performed well with high precision, linearity, and efficient running time than both the manual trypan blue and the flow cytometry methods.
TL;DR: The results suggested that the novel fusion protein antigen successfully constructed by gene SOEing provided the improved diagnostic performance for TB, and other mycobacterial multiepitope fusion proteins may also be worthy of investigation for further enhancing the detection sensitivity.
Abstract: The detection of Mycobacterium tuberculosisspecific antibodies in human sera has been a rapid and important diagnostic aid for tuberculosis (TB) control and prevention. However, any single antigen is not enough to be used to cover the antibody profiles of all TB patients. In this study, a novel fusion protein was constructed using gene splicing by overlap extension (SOEing), and then the antibody level against it in 171 TB patients and 86 controls was evaluated by enzyme-linked immunosorbent assay. Compared with the three individual antigen (16kDa: sensitivity 19.9%, specificity 96.5%; MPT64: sensitivity 75.4%, specificity 34.9%; 38kDa: sensitivity 33.3%, specificity 83.7%), the fusion protein antigen (sensitivity 42.1%, specificity 89.5%) gave the best diagnostic performance with the largest receiver operating characteristic curve area 0.656 (95% confidence interval [CI], 0.590‐0.721; Po0.01). These results suggested that the novel fusion protein antigen successfully constructed by gene SOEing provided the improved diagnostic performance for TB, and other mycobacterial multiepitope fusion proteins may also be worthy of investigation for further enhancing the detection sensitivity. J. Clin. Lab. Anal.
TL;DR: ErythrocyTosis and severe microcytosis, together with a high percentage of microcytes and a moderate increase in IRF, is the profile of β‐thalassemia carriers, whereas anisocytosis and the hypochromic subset correlates with the severity of the anemia in iron‐deficient patients.
Abstract: (23.4%) than in mild cases (12.4%), Po0.0001. %MicroR was more increased in thalassemia (38.6 %) than in the mild IDA (16.5%, Po0.001) and in severe IDA (21.6%, Po0.001). Immature reticulocyte fraction (IRF) mean values in the groups were statistically different; the thalassemia group had an intermediate value (8.7%) between healthy (4.4%) and IDA (16.7 and 12.9%). Conclusions: Erythrocytosis and severe microcytosis, together with a high percentage of microcytes and a moderate increase in IRF, istheprofileofb-thalassemia carriers, whereas anisocytosis and the hypochromic subset correlates with the severity of the anemia in iron-deficient patients. J. Clin. Lab. Anal. 25:223‐228, 2011. r 2011 Wiley-Liss, Inc.
TL;DR: Evaluating the relations of ADMA and NO with the obesity‐linked peptides, such as ghrelin, leptin, and adiponectin in postmenopausal women free of hormone replacement therapy found that estrogen deficiency alone may not cause an increase in ADMA levels unless the women are prone to disturbances in energy homeostasis.
Abstract: Background: It has been reported that estrogen deficiency after menopause might cause a decrement in nitric oxide (NO) bioavailability by increasing the level of asymmetric dimethylarginine (ADMA), a major endogenous nitric oxide synthase inhibitor, thus leading to abnormalities in endothelial function. Because NO plays an important role on feeding behavior, ADMA may be involved in the pathogenesis of obesity, too. This cross-sectional study aimed to evaluate the relations of ADMA and NO with the obesity-linked peptides, such as ghrelin, leptin, and adiponectin in postmenopausal women free of hormone replacement therapy. Methods :A diponectin, ghrelin, leptin, ADMA, and NOx (total nitrite/ nitrate) were measured in 22 obese (BMI: 30‐47kg/m 2 ) and 19 normal weight (BMI: 21.5‐26kg/m 2 ) postmenopausal women. Anthropometric measurements (height, weight, BMI, waist, and hip circumferences) were recorded. Statistics were made by the Mann‐Whitney U-test. Results: Ghrelin and adiponectin levels were significantly lower (Po0.001), whereas ADMA and leptin levels were higher in obese women than in normal weight controls (Po0.01 and 0.001, respectively). BMI was correlated negatively with adiponectin and ghrelin and positively with ADMA and leptin levels. No correlation existed between ADMA and NO. Conclusion: Estrogen deficiency alone may not cause an increase in ADMA levels unless the women are prone to disturbances in energy homeostasis. In spite of the high ADMA levels, the unaltered NO levels in plasma may be owing to ongoing inflammatory conditions. J. Clin. Lab. Anal. 25:174‐178, 2011. r 2011 Wiley-Liss, Inc.
TL;DR: It is found that lung cancer patients produced higher TGF‐β1 levels than healthy individuals, and this study proposes the measurement of serum T GF‐ β1 levels as a complementary diagnostic test in lung cancer detection.
Abstract: Lung cancer is a malignant disease with increasing mortality rates. Cytokines play a role in normal cell growth regulation and differentiation and are also implicated in malignant disease. Among these cytokines, Transforming Growth Factor β type 1 (TGF-β1) acts as a tumor promoter in malignant cells. Several clinical studies have found high levels of TGF-β1 in various cancer types. The aim of this study was to establish a TGF-β1 cut-off point as a complementary diagnostic tool in lung cancer detection. Therefore, 72 clinically well-characterized individuals were studied, 41 lung cancer patients and 31 healthy subjects. Serum TGF-β1 concentration was measured by an enzyme-linked immunosorbent assay (ELISA). We compared statistically the serum TGF-β1 concentration between both groups with analysis of variance, linear regression and receiver operating curve analysis. We observed that lung cancer patients produced higher TGF-β1 levels than healthy individuals (37,225±9,436 vs. 28,416±9,324 pg/ml, P<0.001). The cut-point diagnostic value was 30,500 pg/ml with 80.5% sensitivity, 64.5% specificity and odds ratio: 7.5, 95% CI: 2.6–21.8. Conclusions: We found significantly higher TGF-β1 levels in lung cancer patients than in healthy individuals. We propose the measurement of serum TGF-β1 levels as a complementary diagnostic test in lung cancer detection. J. Clin. Lab. Anal. 25:238–243, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: S serum IgG antibody titer test is useful in the prognosis of periodontitis recurrence during the SPT phase and the recurrence ratio in the high IgG titer group to Gram‐negative obligate anaerobe, Prevotella intermedia, Treponema denticola, and C. rectus was significantly higher than that of the normal IgGTiter group.
Abstract: Chronic periodontitis is associated with systemic diseases such as atherosclerosis. In this study, we evaluated the efficacy of serum IgG antibody titer to periodontal bacteria for prognosis of periodontitis recurrence during supportive periodontal therapy (SPT) phase. The 139 patients during SPT phase were selected and divided to two groups as follows: "Stable" and "Recurrence" group at SPT phase for case-control study: "High IgG titer" and "Normal IgG titer" group before transition to SPT phase for cohort study. We examined whether clinical findings or serum IgG antibody titers to periodontal bacteria are risk factors for the development of periodontitis recurrence. Case-control study showed that there were significant differences between the stable and recurrence groups in age and number of teeth. The serum IgG antibody titer to Eikenella corrodens FDC1073, Porphyromonas gingivalis SU63, and Campylobacter rectus ATCC33238 was significantly higher in the recurrence group. Next, we found, that the recurrence ratio in the high IgG titer group to Gram-negative obligate anaerobe, Prevotella intermedia, Treponema denticola, and C. rectus was significantly higher than that of the normal IgG titer group. Taken together, serum IgG antibody titer test is useful in the prognosis of periodontitis recurrence during the SPT phase.
TL;DR: Urinary cotinine measurement and self‐report of parents' smoking seems to be accurate to assess the exposure to SHS in mass screening, and individual variability of CYP2A6 gene polymorphism should be taken into consideration.
Abstract: The use of a biomarker is mandatory for quantitative analysis of exposure to secondhand smoke (SHS). This article summarizes urinary biomarkers of smoke exposure which can be now quantified. The most reliable urinary biomarkers to assess the exposure to SHS are NNAL 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and NNAL-Glucuronides, which is metabolites of tobacco-specific nitrosamine. These substances were detected even in 50% of children who had undetectable level of cotinine (<0.5 ng/ml). Urinary cotinine, which is determined by a highly sensitive competing enzyme immunoassay, is also a useful biomarker. However, individual variability of CYP2A6 allele,in which nicotine is catalyzed to cotinine, affects the level of urinary cotinine. Approximately 20% of Japanese subjects have homozygotes or heterozygotes of the CYP2A6*4 allele, which has impaired nicotine metabolism and subsequently may underestimate the actual exposure to SHS. In assessing the exposure to SHS, therefore, individual variability of CYP2A6 gene polymorphism should be taken into consideration. The combination of urinary cotinine measurement and self-report of parents' smoking seems to be accurate to assess the exposure to SHS in mass screening. J. Clin. Lab. Anal. 25:354–358, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: Nephelometry is the clinical urinary albumin method with best analytical performance in this study and is selected as the preferred method for clinical use.
Abstract: Background: Microalbuminuria is an indicator of kidney damage and a risk factor for the progression kidney disease, cardiovascular disease, and so on. Therefore, accurate and precise measurement of urinary albumin is critical. However, there are no reference measurement procedures and reference materials for urinary albumin. Methods: Nephelometry, turbidimetry, colloidal gold method, radioimmunoassay, and chemiluminescence immunoassay were performed for methodological evaluation, based on imprecision test, recovery rate, linearity, haemoglobin interference rate, and verified reference interval. Then we tested 40 urine samples from diabetic patients by each method, and compared the result between assays. Results: The results indicate that nephelometry is the method with best analytical performance among the five methods, with an average intraassay coefficient of variation (CV) of 2.6%, an average interassay CV of 1.7%, a mean recovery of 99.6%, a linearity of R=1.00 from 2 to 250 mg/l, and an interference rate of <10% at haemoglobin concentrations of <1.82 g/l. The correlation (r) between assays was from 0.701 to 0.982, and the Bland–Altman plots indicated each assay provided significantly different results from each other. Conclusion: Nephelometry is the clinical urinary albumin method with best analytical performance in our study. J. Clin. Lab. Anal. 25:324–329, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: The determination of IMA levels in pediatric oncology patients with poor prognoses from STSs and NB may play an important role in predicting response to therapy and overall outcome.
Abstract: In this study, the levels of ischemia-modified albumin (IMA) in pediatric oncology patients with soft tissue sarcomas (STSs) and neuroblastoma (NB) were analyzed. To date, there have been no studies concerning IMA in these groups of patients. Ninety-nine children with STSs and NB were analyzed from 2006 to 2009, and 30 healthy children were also enrolled in the study. IMA levels were measured throughout treatment in all patients. The levels of IMA in all cancer patients (mean 116.8±39.3 U/ml), in patients with STSs (mean 119.8±27.5 U/ml), and in patients with NB (mean 114.6±36.6 U/ml) were significantly higher than in the control patients (mean 87.3±38.3 U/ml; P=0.0013, 0.0066, and 0.0164, respectively). IMA levels increased before and during the treatment compared with levels in the controls. The determination of IMA levels in pediatric oncology patients with poor prognoses from STSs and NB may play an important role in predicting response to therapy and overall outcome. J. Clin. Lab. Anal. 25:255–258, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: It may be assumed that during the progression of an inactive hepatitis B carrier to being actively infected, reduced paraoxonase and arylesterase activities may be observed.
Abstract: The aim of this study was to determine and evaluate the activity of paraoxonase and arylesterase enzymes in various clinical forms of hepatitis B infection and to investigate the correlation between these parameters and chronic disease course/fibrosis. Overall, 40 patients diagnosed as hepatitis B carriers (CIHBV), 40 chronic active hepatitis B (CAHBV) patients, and 40 healthy adults (control group) between 18 and 65 years of age were enrolled the study. Serum paraoxonase and arylesterase activities were measured spectrophotometrically. Their activities were significantly lower in patients with CAHBV compared with CIHBV patients or with control group patients (P 0.05). The histology activity index of CAHBV patients did not correlate with paraoxonase and arylesterase activities (P>0.05). In light of these findings, it may be assumed that during the progression of an inactive hepatitis B carrier to being actively infected, reduced paraoxonase and arylesterase activities may be observed. J. Clin. Lab. Anal. 25:311–316, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: Based on the results obtained, replacement of a reliable filter paper for preserving finger‐prick blood samples is a trustable and useful facilitator particularly in remote malaria‐endemic areas.
Abstract: Background: Venipuncture sampling in test tubes for detecting malaria parasites using PCR assays possesses a number of limitations such as reluctance of patients, some difficulties in transportation of blood samples and freezing them for long time. To overcome the mentioned limitations, some approaches have been employed by a number of authors. This study was proposed to compare between DNA Banking Card (DBC) filter papers containing dried finger-prick blood and venipunctured frozen liquid blood. Methods: A total of 75 specimens was prepared from the equal enrolled individuals using three blood storage approaches; making Geimsa-stained thin and thick smears from each individual to determine the malaria-positive or -negative specimens, spotting two to three drops of finger-prick blood onto the DBC filter paper, and collecting a 2-ml venous blood sample into EDTA-contained test tube from each individual. A semi-nested Multiplex PCRtechnique with DNA extracted from the two latter sets of specimens was used for plasmodia diagnosis. Results: DNA samples isolated from dried blood spotted on the DBC filter papers resulted in 32 (42.7%) positive and 43 (57.3%) negative cases comparable with the results outcome of frozen liquid blood with 35 (46.7%) positive and 40 (53.3%) negative cases. Statistical analysis revealed higher sensitivity for SnM-PCR using DNA from liquid blood with 100% vs. dried blood spotted on DBC with 97% but higher specificity for the DBC with 100% vs. liquid blood with 95.2%. Conclusions: Based on the results obtained from this study to overcome the problems of venipuncture frozen liquid blood sampling, replacement of a reliable filter paper for preserving finger-prick blood samples is a trustable and useful facilitator particularly in remote malaria-endemic areas. J. Clin. Lab. Anal. 25:185–190, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: The rapid and accurate diagnostic capability of single‐tube multiplex RT‐PCR used in the study appears to be promising enough to be commonly used for dengue viral detection as well as serotyping.
Abstract: This study has evaluated the clinical applicability of a single-tube multiplex RT-PCR as compared with a two-step nested RT-PCR for the diagnosis as well as serotyping of dengue virus in patient's samples. Seventy-six acute phase blood samples collected from clinically suspected dengue patients during the 2008 outbreak were subjected to two-step nested RT-PCR and single-tube multiplex RT-PCR for dengue diagnosis and serotyping. Of the 76 samples, 17 (22.4%) were positive for dengue viral RNA. Single dengue virus infection was found in 16 cases and 1 had concurrent infection with two serotypes (3&1). Dengue serotype 3 was the predominant serotype (70.5%), followed by serotype 1 (23.5%). Single-tube multiplex PCR had concordant result with that of two-step nested RT-PCR including the one with concomitant infection. This study reveals the predominance of dengue serotype 3 in North India in addition to the co-circulation of multiple serotypes and concomitant infection. The rapid and accurate diagnostic capability of single-tube multiplex RT-PCR used in the study appears to be promising enough to be commonly used for dengue viral detection as well as serotyping.
TL;DR: The findings indicate that the accuracy of RHD gene using three regions (exons 4, 5, and 10) can be sufficient for clinical application in a multi‐ethnic population.
Abstract: Background: Maternal plasma analysis for the determination of the fetal RHD status is an exciting tool for the management of RhD-negative pregnant women, specially sensitized women. We assessed the accuracy of fetal RHD genotyping by analysis of maternal plasma in a multi-ethnic population. Methods: We analyzed plasma samples from 88 RhD-negative pregnant women between 11 and 39 weeks of gestation, median age of 28 years old to determine the fetal RHD genotype. This population was from Southeastern Brazil with high mixed ethnic background. Fourteen patients (16%) had anti-D alloantibody. We used Taqman primers and probes to detect by real-time PCR, exons 4, 5, and 10 of RHD. As internal controls we used primers/probes sets to SRY and CCR5. Peripheral or umbilical cord bloods from respective nenonates were collected during delivery and hemagglutination was performed. Results: Fifty-eight samples (66%) were genotyped as RHD+, 27 samples (31%) showed complete absence of RHD and 3 samples (3 %) presented a D variant (RHDψ). All the results agreed with the neonatal typing, including the three fetuses with the RHDψ, phenotyped as RhD-negative. Thus, the accuracy of the fetal RHD genotyping in this mixed population was 100%. The earliest pregnancy in which fetal RHD was detected was 11 weeks. Conclusion: Our findings indicate that the accuracy of RHD gene using three regions (exons 4, 5, and 10) can be sufficient for clinical application in a multi-ethnic population. This knowledge helped us on the development of a feasible protocol for fetal RHD genotyping on DNA from maternal plasma for our population. J. Clin. Lab. Anal. 25:100–104, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: The sensitivity of HCC diagnosis can be improved by combined detection of AFP and GPC3 mRNA expressions, which is HCC‐specific, and may indicate HCC metastasis.
Abstract: This work aimed to investigate the correlation of the expression of α-fetoprotein (AFP) and glypican-3 (GPC3) mRNAs in the peripheral blood with primary hepatocellular carcinoma (HCC) and HCC metastasis by using multiple fluorescence quantitative reverse transcriptase-polymerase chain reaction (FQ-RT-PCR). Peripheral blood samples from 100 patients with HCC were collected. The positive expression rates of AFP mRNA of HCC, hepatitis B, and cirrhosis patients were 56, 5, and 10%, respectively. AFP mRNA was not detected in healthy subjects, hepatic hemangioma, or hepatic metastasis patients' samples. Those of GPC3 mRNA of HCC patients were 76%. GPC3 mRNA was not detected in healthy subjects, hepatitis B, cirrhosis, hepatic hemangioma, or hepatic metastasis patients' samples. In HCC patients' samples, the combined positive rate of AFP and GPC3 mRNA expressions was 81%. The relative expression levels of GPC3 mRNA in the metastasis group and nonmetastasis group were 0.98±0.38 and 0.72±0.26, respectively, and showed significantly different (P=0.001). However, no significant difference was observed in the AFP mRNA expression levels (P=0.134). In conclusion, the sensitivity of HCC diagnosis can be improved by combined detection of AFP and GPC3 mRNA expressions. GPC3 mRNA is HCC-specific, and may indicate HCC metastasis.
TL;DR: The Hcy level was significantly different according to various factors, especially in the gender and folate level, and the reference interval should be determined for each ethnic population and for each assay.
Abstract: In this study, we estimated the reference intervals of the serum homocysteine (Hcy) level using two automated immunoassays, and we demonstrated the effects of various factors on the Hcy level in a Korean population. We calculated the gender- and assay-specific reference intervals using the data from 809 healthy Koreans, and we assessed the effects of physiologic and lifestyle factors on the Hcy level. The upper limit was higher in males (19.21 and 19.76 μmol/l) than that in females (14.99 μmol/l and 15.16 μmol/l, AxSym and ADVIA centaur, respectively); the upper limits were comparable between the two assays. Smokers, vitamin nonusers, and persons without regular exercise showed a lower folate level and a higher Hcy level. The risk of hyperhomocysteinemia was significantly associated with the male gender (adjusted OR: 5.705, P-value: 0.008) and with the low folate level group (adjusted OR: 10.412, P-value: 0.002) on the multivariate analysis. The Hcy level was significantly different according to various factors, especially in the gender and folate level. The reference interval should be determined for each ethnic population and for each assay. The appropriate cutoff for assessing the risk for cardiovascular disease or stroke should also be validated in each population. J. Clin. Lab. Anal. 25:317–323, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: There is an increased prevalence of IgA antitissue transglutaminase antibodies, which suggests the need to use this method as an effective first‐line test in the screening of celiac disease in children with type 1 diabetes mellitus.
Abstract: The association of celiac disease with type 1 diabetes mellitus is known, but the evolution of celiac disease is most frequently asymptomatic, without any clinical signs. Thus, diagnosis is impossible to make in the absence of serological tests. Our study aimed to determine the prevalence and the efficiency of IgA antitissue transglutaminase antibodies in the screening of celiac disease in children with type 1 diabetes mellitus. Method: During the course of 2008–2009, we performed an analytical clinical study that included the determination of IgA antitissue transglutaminase antibodies in a group of 119 children with type 1 diabetesmellitus. Fifty-seven percent of the subjects were male and 43% were female, with a mean age of 11±4 years. Results: By evaluating IgA antitissue transglutaminase antibodies, we obtained a prevalence of 9.2% in children with type 1 diabetes mellitus, with a sensitivity and specificity of 80 and 82.6%, respectively. Conclusions: There is an increased prevalence of IgA antitissue transglutaminase antibodies, which suggests the need to use this method as an effective first-line test in the screening of celiac disease in children with type 1 diabetes mellitus. J. Clin. Lab. Anal. 25:156–161, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: Red blood cell distribution width values at ≤34 weeks in newborns are higher than at ≥35 weeks, which may be useful in the differential diagnosis of neonatal hematologic diseases together with other red cell parameters.
Abstract: Aim: To evaluate the normal range of red blood cell distribution width (RDW) in term and preterm newborns dependent on gestational age. Material and methods: A total of 1,594 preterm and term neonates were admitted to our neonatology department. Infants were divided into two groups according to their gestational age. Group 1 consisted of infants with ≤34 weeks of gestation; group 2 consisted of infants with ≥35 weeks of gestation. Infants in Groups I and II were subdivided according to their gestational age. Gestational age, birth weight, sex, hemoglobin and hematocrit, MCV levels of all newborns were recorded, and RDW was compared between the groups. Results: A total of 1,594 newbornswere enrolled in the study. Group 1 (≤34 weeks) consisted of 725 newborns and Group 2 (≥35 weeks) consisted of 869 newborns. The mean normal range of RDW in Group 1 was 17.8 ± 2.1 and of group II was 16.7 ± 1.6 (P<0.05). The normal range for RDW values at 32–34 weeks was higher than at 35–36 gestational weeks, and at 37–42 weeks (P = 0.002 and 0.003). Conclusion: RDW values at ≤34 weeks in newborns are higher than at ≥35 weeks. This may be useful in the differential diagnosis of neonatal hematologic diseases together with other red cell parameters. J. Clin. Lab. Anal. 25:422–425, 2011. © 2011 Wiley Periodicals, Inc.
TL;DR: This work attempted to rapidly estimate everolimus concentration from apparent sirolimus concentration obtained by using Architect siro Limus immunoassay and mathematical equations (both polynomial and linear).
Abstract: United States Food and Drug Administration (FDA) in 2010 approved the use of immunosuppressant drug everolimus, which requires therapeutic drug monitoring in whole blood. Taking advantage of structural similarity between sirolimus and everolimus we attempted to rapidly estimate everolimus concentration from apparent sirolimus concentration obtained by using Architect sirolimus immunoassay and mathematical equations (both polynomial and linear). Mathematical equations were derived by curve-fitting methods based on observed apparent sirolimus concentration and true everolimus concentration determined by a liquid chromatography combined with mass spectrometry (LC/MS) method using eight everolimus standards (concentration range 1–30 ng/mL) prepared in whole blood. In order to determine the validity of our approach, we analyzed 12 specimens from patients receiving everolimus using both Architect sirolimus assay and LC/MS method. We observed good correlation between calculated everolimus values and true everolimus values as determined by LC/MS. However, if a patient is switched from sirolimus to everolimus, then sirolimus immunoassay can roughly estimate everolimus concentration plus any residual sirolimus present in whole blood and it is not possible to calculate everolimus concentration. J. Clin. Lab. Anal. 25:207–211, 2011. © 2011 Wiley-Liss, Inc.
TL;DR: According to the results, flow cytometric analysis of leukocyte surface antigen expressions should be performed using +4°C temperature throughout the process and within 6 hr of collection or after 6 or 24hr storage.
Abstract: Flow cytometric analysis of leukocyte surface antigens has been used to characterize infectious and septic processes in patients. We wanted to investigate how the sampling and processing temperature, the anticoagulant used, and the storage of the sample influence leukocyte immunophenotyping. Four blood samples, two using acid citrate dextrose and two using heparin as an anticoagulant, were taken from five intensive-care unit patients with severe sepsis and five healthy volunteers. The samples were collected, stored, and processed either at +4°C or at room temperature (RT). The samples were processed for flow cytometric analysis within 1 hr of collection or after 6 or 24 hr storage. The surface antigens of interest were neutrophilic CD11b and CD64, monocytic CD11b, CD14, CD40, CD64, CD80 and HLA-DR, and lymphocytic CD69 (separately in CD4+ and CD8+ T cells, B cells, and natural killer cells). The fluorescence intensities were higher at RT than at +4°C. During storage the intensities increased at RT, but at +4°C there were only minor changes. The effects were similar with both anticoagulants studied. According to our results, flow cytometric analysis of leukocyte surface antigen expressions should be performed using +4°C temperature throughout the process and within 6 hr.