Setting Up and Statistical Evaluation of a New Haemoglobin Assay (original) (raw)
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Novel method for screening for the presence of haemoglobin S in blood
Transfusion Medicine, 2005
A high-performance liquid chromatography (HPLC) method designed for the measurement of haemoglobin (Hb)A 1c in blood was investigated for use as a screening method for the presence of HbS in blood. In the Bio-Rad VARIANT II HbA 1c method, HbS was found to have a specific retention time and percentage Hb value that enabled the detection of HbS in blood. Other Hb variants did not have the same combination of retention time and percentage Hb as HbS. The HPLC method was superior to the HbS solubility test in ease of performance and readability. Also, the proposed method showed less interference than the solubility test and could be used with samples from all age groups. The proposed method takes 3 min per sample to perform and is thus suitable for large-scale screening.
Evaluation of three methods for hemoglobin measurement in a blood donor setting
Sao Paulo Medical Journal, 1999
International Committee for Standardization in Hematology (ICSH) method. The azide metahemoglobin is measured at 570 nm. To compensate for turbidity, e.g. due to lipids, a second measurement is taken at 880 nm. 3 The advantage of this technology is that it is simple, rapid and does not require sophisticated hematological equipment. The system is designed to use capillary, venous or arterial blood. Also, the instrument is small and portable ABSTRACT Context: The hemoglobin (Hb) level is the most-used parameter for screening blood donors for the presence of anemia, one of the most-used methods for measuring Hb levels is based on photometric detection of cyanmetahemoglobin, as an alternative to this technology, HemoCue has developed a photometric method based on the determination of azide metahemoglobin. Objective: To evaluate the performance of three methods for hemoglobin (Hb) determination in a blood bank setting. Design: Prospective study utilizing blood samples to compare methods for Hb determination. Setting: Hemotherapy Service of the Hospital Israelita Albert Einstein, a private institution in the tertiary health care system. Sample: Serial blood samples were collected from 259 individuals during the period from March to June 1996. Main Measurements: Test performances and their comparisons were assessed by the analysis of coefficients of variation (CV), linear regression and mean differences. Results: The CV for the three methods were: Coulter 0.68%, Cobas 0.82% and HemoCue 0.69%. There was no difference between the mean Hb determination for the three methods (p>0.05). The Coulter and Cobas methods showed the best agreement and the HemoCue method gave a lower Hb determination when compared to both the Coulter and Cobas methods. However, pairs of methods involving the HemoCue seem to have narrower limits of agreement (± 0.78 and ± 1.02) than the Coulter and Cobas combination (± 1.13). Conclusion: The three methods provide good agreement for hemoglobin determination.
Background: Population-based surveys matched by time and in the same country, but which use different methodologies for determining hemoglobin (Hb) concentration, have shown inconsistencies in estimating anemia prevalence. This study aims to estimate measurement errors in Hb using the HemoCue 201+ in capillary blood, both in drops and pooled, compared with venous blood analyzed both with the HemoCue 201+ and clinical methodologies as references. Methods: 149 participants (children, women, and older adults) interviewed in their homes in Jojutla, Morelos, Mexico, were randomly allocated to donate either a drop of capillary blood or 350 µL of pooled capillary blood. Each group also donated 4 mL of venous blood. Hb was analyzed in duplicate using the HemoCue 201+ and by a reference method using cyanmethemoglobin. We also compared the latter method with a clinical hemocounter. We performed Bland Altman and Lyn’s concordance analyses. We calculated 95% confidence intervals (±95% CI).Resul...
Purification and characterization of human hemoglobin: effect of the hemolysis conditions
International Journal of Biological Macromolecules, 2004
Human hemoglobin was isolated and purified by anion exchange chromatography. To isolate hemoglobin, outdated red blood cells (RBC) were transformed into carbonylhemoglobin, by reaction with carbon monoxide, and submitted to washing/centrifugation procedures, to eliminate other plasma proteins. Albumin was quantified in each supernatant, by the bromcresol green method. Hemolysis was performed in three different hypotonic media (water, 0.01 M NaCl and 5 mM Tris/HCl buffer at pH 7.4), at 8 • C for 24 h. Sonication for 5 min was also used to lyse RBC. After isolation of hemoglobin, additional purification was carried out by anion exchange chromatography on AG MP-1, Q-SFF and both exchangers. Hemoglobin concentration of hemolysates and of purified solutions were determined by the hemiglobincyanide method. Residual phospholipids were extracted from the four different hemolysates, as well as from the purified hemoglobin solutions, and were analyzed by high performance liquid chromatography. Native and SDS-polyacrylamide gel electrophoresis experiments were performed on purified hemoglobin samples to verify the presence of proteins other than hemoglobin. According to the results, the hemolysis conditions have influence on the purification of hemoglobin.
Validity of Estimation of Haemoglobin Content in Dried Blood Spot Samples
Indian Journal of Hematology and Blood Transfusion, 2017
The ability to use indirect cyanmethemoglobin method using 20 ll of dried blood spots (DBSs) on filter paper for the analysis of haemoglobin (Hb) levels could be an important diagnostic tool for areas that have limited access to laboratory facilities. We assessed the validity of a method for Hb estimation in which a single drop of whole blood was directly taken on the filter paper. We collected 124 DBSs containing 20 ll of blood (filter paper A) and single drop of whole blood (filter paper B) from subjects living in Nainital, Uttrakhand. Estimation of Hb was done by indirect cyanmethemoglobin method in both the filter papers. A correction factor for predicting value of Hb from DBSs of single drop of whole blood was established. The Bland-Altman plot suggested that the difference in Hb values obtained by the single drop of blood and 20 ll of DBSs was within the 1.5 SD limits, suggesting high validity of the correction factor. The estimation of Hb using single drop of whole blood on filter paper after applying the correction factor provides results similar to indirect cyanmethemoglobin method using 20 ll of blood. Hence, single drop of whole blood on filter paper can be used as an alternate method for estimation of Hb in large scale community surveys.
Analytical Biochemistry, 1979
A method for determining the concentration of available haptoglobin in plasma and other solutions is described. The procedure involves absorbance measurements in a mixture of plasma and hemoglobin before and after the addition of sodium dithionite. The haptoglobin content of the solution is calculated from the ratio of the absorbances at the Soret peaks of the oxy-and deoxyhemoglobin. The assay can be performed in minutes, and the error of measurement is well under 10%.
Accuracy and Precision of Hemoglobin Determination
The Journal of Nutrition, 2024
Background: Anemia prevalence estimates reported in population surveys can vary based on the blood specimen source (capillary or venous) and analytic device (hematology autoanalyzers or portable hemoglobinometers) used for hemoglobin (Hb) determination. Objectives: This study aimed to compare accuracy and precision of Hb measurement in three blood specimen types on three models of hemoglobinometers against the results from venous blood from the same individuals measured on automated analyzers (AAs). Methods: This multisite (Cambodia, Ethiopia, Guatemala, Lebanon, Nigeria, and Tanzania) study assessed Hb measurements in paired venous and capillary blood specimens from apparently healthy women (aged 15-49 y) and children (aged 12-59 mo) using three Hemo-Cue® Hb models (201þ, 301, and 801). Measurements were compared against reference values: venous blood in hematology AA and adjusted via regression calibration or mean difference in HemoCue® Hb. Venous, capillary pool, and single-drop capillary blood specimens were assessed for accuracy and precision. Results: Venous blood measured using HemoCue® Hb 301 exhibited a positive mean error, whereas responses in HemoCue® Hb 201þ and 801 were nondirectional compared with the reference. Adjustment with the reference harmonized mean errors for all devices across study sites to <1.0 g/L using venous blood. Precision was highest for venous blood (AE5-16 g/L) in all sites, lowest for single-drop capillary (AE9-37 g/L), and intermediate (AE9-28 g/L) for capillary pool blood specimen. Imprecision differed across sites, especially with both capillary blood specimens, suggesting different levels of personnel skills. Conclusions: Findings suggest that venous blood is needed for accurate and precise Hb determination. Single-drop capillary blood use should be discouraged owing to high measurement variability. Further research should evaluate the viability and reliability of capillary pool blood for this purpose. Accuracy of HemoCue® Hb devices can be improved via standardization against results from venous blood assessed using AA.
Journal of diabetes and metabolic disorders, 2011
Background: Main objective was to measure glycated hemoglobin (HbA1c) in dried blood spots on paper filter and in whole blood samples in diabetic patients to evaluate relationship between two methods and their respective reliability. Methods: The 20×10 µl of venous blood samples of 33 diabetics were blotted onto the filter paper allowed to dry at room temperature and then stored at 25˚C and 4˚C. HbA1c was measured via the Turbidimetric Inhibition Immunoassay Technique. The relation was evaluated with correlation and linear regression tests using STATA software and SPSS. Agreement between the results obtained from the dried blood spots and others was evaluated using the Bland and Altman. The pitman's permutation test was also employed to compare the difference in variance. Results: A high positive correlation was detected between whole blood samples and dried blood spots stored at 4˚C (r 2 =0.90) and at 25˚C (r 2 = 0.95). The Bland and Altman graphs, as well as the Pitman tests, ...
Evaluation of hemoglobin estimation with non-cyanide alkaline haematin D- 575 method
International Journal of Research in Medical Sciences, 2016
Background: Anemia is serious cause for concern in the world as it impacts on psychological and physical development, behavior and work performance. There are various methods recommended for estimation of hemoglobin for detection of anemia. Each method has its advantages and disadvantages. Aims and objectives of the present study were to compare conventional Hemoglobinocynide (HiCN) method containing potassium ferricyanide and potassium or sodium cyanide with non-cyanide alkaline haematin Method for hemoglobin estimation. Methods: The prospective study was conducted to evaluate the performance of two methods-HiCN method with Drabkin"s reagent and non-cyanide alkaline haematin method with AHD 575 reagent, for hemoglobin determination utilizing 201 blood samples. The data statistically analyzed by using coefficient of variation (CV), linear regression and mean differences. Results: A good correlation was observed for hemoglobin estimation between the HiCN method and non-cyanide alkaline haematin Method with AHD 575 reagent. The correlation coefficient of r= 0.9998 was statistically significant. Conclusions: It was concluded that both methods are accurate and precise, however the toxic and biohazardous effects of potassium ferricyanide and sodium cyanide in HiCN method can be prevented by using alkaline haematin method with AHD 575 reagent.