Open Access Original Research Article

Glycated Hemoglobin Evaluation by Means of Ion Exchange Chromatography and Immunoassay in Normal and Hemoglobinopathy Patients

Shyamali Pal

International Blood Research & Reviews, Page 1-9
DOI: 10.9734/IBRR/2016/24410

Aims and Objectives: High performance liquid chromatography which is basically an Ion Exchange Chromatography, is the most popular as well as expensive method for glycated hemoglobin estimation but biased results have been observed in hemoglobinopathies. Sometimes results cannot be reported also. In such situation the laboratory adopted immunoassay method and successfully reported the results. Therefore, a comparative analysis of glycated hemoglobin results of normal and abnormal hemoglobin were essential as high performance liquid chromatography is the most sought after method.

Study Design: Number of patients with normal adult hemoglobin compared were 120. Patients with Hb-AE thalassemia were 21. Patients with β-thalassemia trait were 36. Hb-E-β thalassemia patients were 6, β-thalassemia homozygous 10 and HbSS patients tested were 4. The glycated hemoglobin values obtained were from euglycemic to severe diabetic range. The instruments used for measurement are D10 BIORAD and Cobas Integra 400plus.

Data Analysis: Deviations calculated from results of same patient obtained from two system. Regression coefficients of each group were calculated. Trend analysis was done and graphically presented.

Results and Discussion: Regression analysis of normal, HbAE&β-variant patients show coefficients are 0.988,0.976 &0.961 respectively proving excellent comparability of two methods. But trend analysis of deviations show right shift of 85%HbAE& 86.1%β-variants .Similar bias was observed in external quality assessment results also. The immunoassay results were comparable with boronate affinity chromatography and enzymatic estimation method. The deviation in HPLC observed only when in external assessment sample a window was seen. Therefore, observations of the trend of patients and EQAS sample indicates low bias of HbA1c estimation by HPLC method in D10.In severe forms of Hemoglobinopathy i.e. HbSS, Hb-E-β and β-thalassemia homozygous 75%,67% & 60% results could not be obtained from D10. The acceptability of immunoassay results checked by comparing glucose fasting and estimated average glucose calculation values.

Conclusion: Immunoassay, the second popular method is reliable, less expensive method and results remain unaffected in hemoglobinopathies.


Open Access Original Research Article

Detection of Prethrombin 1 in Human Blood Plasma

Dar’ya Korolova, Volodymyr Chernyshenko, Tetiana Platonova, Tamara Chernyshenko, Eduard Lugovskoy

International Blood Research & Reviews, Page 1-7
DOI: 10.9734/IBRR/2016/24683

Background: Prethrombin 1 is one of prothrombin derivatives which appears in the presence of thrombin in vitro. However, existence of prethrombin 1 in vivo remained questionable. The aim of present work was to detect of prethrombin 1 in vivo at abdominal aortic aneurysm, hip replacement after fracture and stroke.

Methodology: Blood plasma samples of patients with abdominal aortic aneurysm, hip replacement after fracture, stroke, and patients which were treated with warfarine were collected. Detection of prethrombin 1 was completed using three independent approaches: combination of prothrombin index and ecamulin (ecarin) index; the APTT-test modified by addition of exogenous prothrombin; Western-blotting of blood plasma using polyclonal antibody to prothrombin.

Results: Prethrombin 1 presence in patients’ blood plasma at studied pathologies was proven. Inactive prothrombin (descarboxy-prothrombin), but not prethrombin 1 was found in blood plasma of patients which were treated with warfarine. Developed approach can be used for testing the prethrombin 1 in clinical diagnostics.

Conclusion: Prethrombin 1 at different pathologies, accompanied by coagulation disorders was detected in patients’ blood plasma directly.

Open Access Original Research Article

Changes in Packed Cell Volume and Some Biochemical Parameters of Adult Females during Pregnancy in North-West Nigeria

Imoru Momodu, Isah Suleiman Yahaya

International Blood Research & Reviews, Page 1-6
DOI: 10.9734/IBRR/2016/24544

Background: Pregnancy is associated with changes in woman’s renal function, carbohydrate and protein metabolism and particularly hormonal pattern. The objective of this study was to determine the values of packed cell volume and some biochemical parameters during pregnancy in North-west Nigeria due to scanty information documented in this part of Nigeria.

Materials and Methods: One hundred and fifty apparently healthy pregnant women, aged 17-40 years, were studied between August, 2010 and October, 2011 at Aminu Kano Teaching Hospital, Kano while 100 age-matched and apparently healthy, non-pregnant women, resident in Kano metropolis as the pregnant women, were used as controls. Blood samples collected from the subjects were analysed for packed cell volume (PCV), random blood sugar (RBS), urea and creatinine using standard laboratory techniques.

Results: There were significantly lower values of packed cell volume and random blood sugar in pregnant women compared to non-pregnant women (P<0.05) and higher value of urea level in pregnant women compared to non-pregnant women (P<0.05).The values of creatinine levels in pregnant and non- pregnant women showed no significant difference (P>0.05) while parity and maternal age showed no effects on the PCV levels during pregnancy.

Conclusion: This study has shown reduced PCV value in pregnant women probably due to haemodilution and dietary deficiency of iron while the lower value of RBS may be attributed to reduced consumption of cassava or carbohydrate rich foods in this environment. Elevated urea level in the studied pregnant women might be associated with their high protein diets.


Open Access Original Research Article

Serum Levels of Gamma-interferon and Interleukin-4 in Homozygous Sickle Cell Anaemia Patients

Omotola T. Ojo, Wuraola A. Shokunbi, Ajayi A. Ibijola, Ganiyu A. Arinola, Philip O. Olatunji, Akeem O. Lasisi, Ayorinde F. Fayehun

International Blood Research & Reviews, Page 1-7
DOI: 10.9734/IBRR/2016/24755

Background: Impaired leucocytes functions is among mechanisms that have been reported to account for the immunocompromised state of patients with sickle cell disease.

Objective: This study assessed cellular immunity using serum IFN-γ and IL-4 levels in patients with sickle cell anaemia (SCA).

Methods: The study comprised of 40 sickle cell anaemia patients in steady state (asymptomatic for at least 4 weeks) and 40 age and sex-matched healthy HbA control. Serum IFN-γ and IL-4 was determined by Enzyme linked immunosorbent assay (ELISA) as described by the manufacturer of the kit.

Results: There was a significant increase in the IFN-γ level in sickle cell anaemia patients in steady state (median value 86.1 pg/ml) compared with HbA controls (median value 55.8 pg/ml) (p=0.04). However, there was no significant difference in the median values of IL-4 level between the HbS (homozygous inheritance of sickle gene) patients and the control subjects (IL-4: p=0.42).

Conclusion: High value of IFN- γ may contribute to inflammation and tissue damage in HbS patients, thus worsening morbidity and mortality.


Open Access Original Research Article

Determination of Platelet and White Blood Cell Counts from Peripheral Blood Smear: An Indispensable Method in Under-resourced Laboratories

Imoru Momodu

International Blood Research & Reviews, Page 1-7
DOI: 10.9734/IBRR/2016/25105

Objective: Blood smear examination serves as a quality control tool in verifying the results generated by the automated analyzer and identification of abnormal or immature cells amongst other functions. This study was undertaken to estimate platelet and White Blood Cell (WBC) counts from Peripheral Blood Smear (PBS) and to correlate them with the results from automated method.

Materials and Methods: Fifty blood samples collected into K3 EDTA from 30 males and 20 females whose ages were from 2 to 50 years, and attended General Out-Patient Department (GOPD) of Aminu Kano Teaching Hospital, Kano between May and November, 2015 were considered for the study. Each blood sample was used for the determination of full blood count using Swelab Alfa hematology analyzer, and preparation of stained blood films using standard laboratory methods.

Results: There were significantly lower values of platelet count (using multiplication factor of 15.0 x109/L) and white blood cell count (using multiplication factor of 2.0 x109/L) to derive (22.42±60.77) x109/L and (4.49±1.04) x109/L by manual (PBS) method as compared to (267.86±77.28) x 109/L and (5.86±1.36) x109/L respectively, for automated method (P<0.001). However, there was no significant difference in estimated platelet count by manual (PBS) method using multiplication factor of 20.0 x109/L compared to automated method (P>0.05). Fairly strong positive correlations were observed for platelet and white blood cell counts when manual method was compared to automated method (r= 0.6828 - 0.7321, P<0.05).

Conclusion: It can be concluded that multiplications factors of 20.0 x109/L per 100X, objective lens and 2.7 x109/L per 40X, objective lens can be used for average numbers of platelets and white blood cells respectively to estimate platelet and white blood cell counts from PBS as the results are comparable to that of the hematology analyzer.