Time Of Serum Storage Research Articles

THE EFFECT OF TEMPERATURE AND STORAGE TIME SERUM ON C- RECATIVE PROTEIN ON PATIENTS WITH TYPE 2 DIABETES MELITUS

C-Reactive Protein is an acute phase protein that is used to determine inflammation. In patients with type 2 diabetes mellitus, increased CRP levels can result in oxidative stress. Stability of serum stored at 2-8℃ stable within 48 hours if the sample needs to be stored for no more than 3 weeks it can be stored for 4℃, if stored at room temperature can be stored for no more than 7 days, but storage is still found serum that is not feasible because the temperature conditions do not meet the requirements. The research used was an experimental study with a sample population of 10 people with type 2 diabetes mellitus who were over 50 years old and had blood sugar ≥ 250 mg/dL and had diabetes for ≥5 years. Serum was examined qualitatively and a semi-quantitative was carried out to determine CRP levels. Serum storage at refrigerator and room temperature for 0, 12, 24, 36, and 72 hours. There was no statistical effect on serum storage time at refrigerator temperature for 0, 12, 24, 36 and 72 hours on CRP titers, there was a statistical effect on serum storage time at room temperature for 36 and 72 hours on CRP titers previously had a titer of 24 mg/L to 12 mg/L. There was no statistical effect on serum storage time at refrigerator temperature on CRP titers, there was a statistical effect on serum storage time at room temperature for 36 and 72 hours on CRP titers.

CHOD-PAP Method Total Cholesterol Examination Validation on Samples Stored for 1 Week In 4-8°C Temperature

High total cholesterol levels cause various diseases, especially those related to the heart and blood vessels. Total cholesterol test uses serum as a sample and must be analyzed within one hour after sampling.However, in the field sometimes there are inspection delays.Some studies have proven that serum stored for one week at a temperature of 4-8°C has different results in total cholesterol examination. For this reason, this study aims to determine the validity of total cholesterol examination using the CHOD-PAP method of serum samples stored for one week at a temperature of 4o-8oC. Validation test based on eight parameters, there are precision, accuracy, linearity, reportable range,limit detection, quantitation limit, recovery, and interference. Only the recovery test was not accepted from eight parameters. Based on the Kruskal Wallis test, the sig value is 0.172 0.05, it can be stated that there is no difference in serum storage time for one week at a temperature of 4-8℃ on total cholesterol levels using CHOD-PAP method.

Improved sensitivity of serum/plasma 1α,25-dihydroxyvitamin D quantification by DAPTAD derivatization

BackgroundAlthough immunoassays have several limitations such as the cross-reactivities of antibodies, such techniques are widely used for serum/plasma 1,25(OH)2D quantification. An accurate method is required for the determination of the 1,25(OH)2D status. MethodsWe designed a serum/plasma 1,25(OH)2D quantification method using LC-MS/MS. Immunoaffinity extraction (IE) and the recently developed Cookson-type reagent 4-(4′-dimethylaminophenyl)-1,2,4-triazoline-3,5-dione (DAPTAD) were used for sample preparation and derivatization, respectively. Analytical and pre-analytical validations were performed. Serum 1,25(OH)2D3 concentrations were determined in 232 healthy Japanese individuals. ResultsThe intra- and inter-assay CVs for 1,25(OH)2D3 were 5.2% and 7.0%, respectively. The limit of quantification for 1,25(OH)2D3 was 7.1pg/ml. Rheumatoid factor (RF) at concentrations below 517IU/ml did not affect serum 1,25(OH)2D analysis. No significant differences were observed for various blood collection tubes, repeated freeze–thaw cycles, whole blood standing time, or serum storage time. A strong correlation between LC-MS/MS and radioimmunoassay (RIA) was observed (r=0.786), but serum 1,25(OH)2D concentrations obtained from RIA were 2-fold higher than those obtained from LC-MS/MS. Serum 1,25(OH)2D3 concentrations by LC-MS/MS were 18.7–53.9pg/ml. ConclusionA highly sensitive and selective LC-MS/MS-based serum/plasma 1,25(OH)2D quantification method was developed using IE and DAPTAD derivatization. This method will enable the accurate determination of serum/plasma 1,25(OH)2D concentrations in the clinical setting.

Stability of total proteins and their electrophoretic fractions in goat serum (Capra hircus), maintained under different condition

The aim of the present study was to evaluate the effect of different storage time and temperature conditions on the concentration of serum total proteins and their electrophoretic fractions in goats. Blood samples were collected from 12 female of Girgentana goats and the obtained serum was divided into 4 aliquots. The first aliquot was analyzed immediately after separation (T0); the second was refrigerated at 4°C and analyzed after 24 (T1), 48 (T2), and 72 (T3) hours; the third was stored at −20°C immediately after centrifugation and analyzed after 2days (T4); and the last aliquot was stored at −20°C after 24h refrigeration and analyzed after 1 week (T5). The serum total proteins, serum protein electrophoretic fractions (albumin, α-, β1-, β2- and γ-globulins) and albumin/globulin ratio (A/G) values were determined. A significant effect of storage conditions on serum total proteins, albumin, α1-, β2- and γ-globulins values was found (P<0.05). The concentration of serum total proteins was significantly higher at T1 and T4 than T0. Albumin showed higher concentrations at T1, T2, T3, T4 and T5 in respect to the values measured at T0. The concentration of α1-globulins was significantly higher at T4 than T0; the β2-globulins values were lower at T3 and T5 respect to T0, whereas the γ-globulins values were lower at T5 than T0. Together these results showed that temperature and time of serum storage are important pre-analytical factors which affect concentrations of serum total proteins and their electrophoretic fractions in electrophoretic analysis. The results of present study suggest that fresh serum samples or samples refrigerated for 24h are recommended for electrophoretic analysis to obtain accurate data, which can help clinicians to differentiate between sick and healthy animals.

Risk Factors for Hepatocellular Carcinoma in a Japanese Population: A Nested Case-Control Study

Epidemiologic studies have shown effects of lifestyle-related factors on risk for hepatocellular carcinoma. However, few cohort studies have incorporated, in a strict and in-depth manner, hepatitis B virus (HBV) and hepatitis C virus (HCV) infections or investigated synergism between such factors. We conducted a nested case-control study using sera stored before hepatocellular carcinoma diagnosis in the longitudinal cohort of atomic bomb survivors. The study included 224 hepatocellular carcinoma cases and 644 controls that were matched to the cases on gender, age, city, time of serum storage, and method of serum storage, and countermatched on radiation dose. Univariate analysis showed that HBV and HCV infections, alcohol consumption, smoking habit, body mass index (BMI), and diabetes mellitus were associated with increased hepatocellular carcinoma risk, whereas coffee drinking was associated with decreased hepatocellular carcinoma risk. Multivariate relative risks of hepatocellular carcinoma (95% confidence interval) were 45.8 (15.2-138), 101 (38.7-263), 70.7 (8.3-601), 4.36 (1.48-13.0), and 4.57 (1.85-11.3), for HBV infection alone, HCV infection alone, both HBV and HCV infections, alcohol consumption of > or =40 g of ethanol per day, and BMI of >25.0 kg/m(2) 10 years before diagnosis, respectively. HBV and HCV infection and BMI of >25.0 kg/m(2) remained independent risk factors even after adjusting for severity of liver fibrosis. Among HCV-infected individuals, the relative risk of hepatocellular carcinoma for a 1 kg/m(2) increase in BMI was 1.39 (P = 0.003). To limit the risk for hepatocellular carcinoma, control of excess weight may be crucial for individuals with chronic liver disease, especially those with chronic hepatitis C.

Factors associated with elevated serum concentrations of anti‐TPO antibodies in subjects with and without diffuse goitre. Results from the Ukrainian–American Cohort Study of Thyroid Cancer and Other Thyroid Diseases Following the Chornobyl Accident

To examine factors associated with the prevalence of elevated anti-thyroid peroxidase antibodies (ATPO) among iodine-deficient adolescents and young adults and test whether associations vary according to the presence of diffuse goitre. Subjects were members of the Ukrainian-American Cohort Study exposed to the Chornobyl accident whose (131)I thyroid dose estimates were below 0.2 Gy. The odds ratios (ORs) for ATPO above 60 U/ml were estimated using logistic regression models for a number of factors in the total population (N = 5133), and separately for thyroid disease-free subjects (N = 3875), those with diffuse goitre (N = 921), and diffuse goitre without autoimmune thyroiditis (AIT; N = 883). Elevated ATPO was found in 9.9% of the total population and ORs were significantly higher in females, older individuals, those examined in earlier calendar years, residents of Kyiv and Chernihiv oblasts, subjects with a family history of thyroid disease, higher thyroid ultrasound volume, suppressed or elevated TSH, blood collection in March to May, very low thyroglobulin (Tg), and shorter serum storage time. When thyroid disease-free subjects and those with diffuse goitre were compared, there were few differences in antibody prevalence, and after excluding individuals with AIT, the only difference was an increased prevalence of elevated ATPO at low urinary iodine in those with goitre alone. Although a number of factors are associated with the prevalence of elevated ATPO in our study group, with the exception of urinary iodine these factors are independent of goitre, and differences between thyroid disease-free subjects and those with diffuse goitre are largely due to AIT.

Hepatitis C virus in autoimmune liver disease in the UK: aetiological agent or artefact?

Hepatitis C virus antibody titres (anti-HCV) were measured in serum from 122 patients with autoimmune liver disease (96 with primary biliary cirrhosis and 26 with autoimmune chronic active hepatitis using three generations of enzyme immunoassay (EIA): first generation--Ortho, EIA1; second generation--Abbott, EIA2; and third generation--Murex, EIA3. Anti-HCV was below the positive cut-off level in all 26 autoimmune chronic active hepatitis patients for all tests, while seropositivity values in primary biliary cirrhosis were 31% (EIA1), 14% (EIA2), and 0% (EIA3). In primary biliary cirrhosis, anti-HCV values as measured by all three tests correlated positively with serum IgG concentrations, serum storage time, and a number of other indices of hepatic dysfunction. Multiple regression analysis showed that anti-HCV values were independently affected by both serum IgG and the length of storage time, although the magnitude of the effects varied between tests. When all three multiple regression models were applied to an extreme clinical example, however, EIA3 was least likely to give a false-positive result. The difference in test performance was emphasised further by examination of anti-HCV values in nine primary biliary cirrhosis patients (confirmed negative by recombinant immunoblot assay 2) in whom serial samples were tested (seven to 14 per patient, stored for one to 138 months). Apparent anti-HCV values (EIA1 and EIA2) increased with increasing serum storage time, but were unchanged when measured by EIA3. A similar pattern was evident in a limited study of five autoimmune chronic active hepatitis patients. As the second generation EIA is in widespread use and confirmatory testing is not always available, the effect of serum storage in addition to hyperglobulinaemia should be considered in the interpretation of positive results in auto immune and in other types of chronic liver disease.

Alpha1-antitrypsin quantitation and Pi typing of stored postmortem blood.

Correlation of necropsy findings with protease inhibitor levels and phenotype is sometimes desirable. This study was designed to determine the feasibility of postmortem protease inhibitor assessment. One hundred fifteen consecutive postmortem samples, stored at -20 C for 24 to 53 months, were analyzed. The time from death to necropsy, storage time, and the pH values of the sera were correlated with alpha1-antitrypsin levels, trypsin inhibitory capacity, and Pi typability. The alpha1-antrypsin level and trypsin inhibitory capacity were not significantly correlated with morgue time, serum storage time, or pH, and mean values were within the expected ranges. A significant decrease in Pi typability occurred when pH was less than 7.0. Moreover, while most (86%) of the sera stored for 2 to 2 1/2 years were typable, only 30% of those stored for more than four years were typable. Determination of alpha1-antitrypsin and trypsin inhibitory capacity are possible with the use of stored postmortem blood. Pi typing is usually possible, provided sera are not acidic and are examined within 2 1/2 years.