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Background: Reticulocytes are young or immature red blood cells released from bone marrow and that contain remanants of ribonucleic acid (RNA) and ribosomes. Reticulocyte count (RC) is the index of erythropoietic activity within bone marrow. The reticulocyte counting methods at clinical laboratories are currently divided into manual and automated.Methods: A total of 500 samples of study cases were processed by manual method using New Methylene Blue (NMB) and automated method based on flowcytometry by PENTRA XLR HORIBA hematology analyzer. All quality control parameters were evaluated and values obtained by both methods were compared using various statistical methods.Results: Automated hematology analyzer provides excellent precision and linearity with no significant carryover. On comparing manual and automated RC method good method correlation was found (correlation coefficient r-0.865), however individual case wise percent deviation between manual and automated RC and CRC varied significantly. In addition within run precision calculated for automated RC differed significantly from manual count. The mean of difference between duplicate readings (150 samples) of manual and automated RC (<5%) were 0.3 and 0.01 respectively while 6.3 and 0.15 respectively for >5% RC. Thus, automated method was found to be more precise than the manual RC.Conclusions: The manual count method for RC associated with significant imprecision compared to flowcytometric method mostly based on interobserver variation and the smaller number of cell being counted. In contrast, the automated method is rapid, easy to operate, count higher number of cells with precise measurement.
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Background: Hemoglobin (Hb) estimation is one of the most frequently ordered investigations. Estimation of exact levels is important to stratify the grade of anemia and subsequently direct the necessary treatment. Conventionally, Hb has been estimated using colorimetric method, which is time tested and recommended by the WHO. Now-a-days, the automated method is also becoming a popular method in many hospitals. However, there are not many studies assessing the accuracy of the automated method over the gold standard. Materials and methods: We retrospectively analyzed hemoglobin values in 180 adult patient-samples (18 per batch in 10 batches). Hemolyzed samples were excluded from the study. Blood samples were drawn in vials having K3 EDTA anticoagulants. After a proper mixing, hemoglobin was estimated by automated Sysmex XS-800i. Parallel estimation for Hb content was done manually by spectrophotometer 4010. Results: Patients ranged from ages 20 to 40 years (M:F=102:78). The lowest value recorded by Sysmex XS-800i was 5.8 while the highest value recorded was 18.6 gm%. The mean hemoglobin concentration on Sysmex XS-800i was 12.89. The lowest, highest and mean values recorded by the cyanmethemoglobin method on photometer were 5, 18 and 13.49 gm% respectively. This showed a mean difference of 0.597 and with significant p-value of <0.001. Conclusion: The lowest values of Hb were similar in both the methods but the mean as well as the highest values differed significantly. Our study found an accuracy of 95.57% with the 5 part analyzer when compared to the gold standard colorimetric method.
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Objective:To compare the results of platelet count done by peripheral smear method and by automated cell counter. Material and Methods: This cross sectional study was conducted in the hematology laboratory of police Hospital Jammu, J & K for the period of two months (February to March 2018). 100 random samples were processed by the automated method (Rayto Haemarray 83) and the manual method (leishman's stained thin blood films) simultaneously. Results: 9 The mean platelet count estimated by the manual method was 208.2 ±100.9× 10 /L, while that estimated by the automated method was 9 206.5 ± 106.0 × 10 /L, with no significant statistical difference between both means (p>0.05). The Pearson correlation test showed significant positive correlation between both methods (r: 0.904), this correlation remained significant when the samples of normal count by the two methods were correlated (r: 0.890), but it was insignificant negative correlation when the samples of low or high counts by the two methods were correlated (r: -0.096 and r: -0.239) respectively. Conclusion: Platelet estimate is an important step in assessment of platelet count and it should be done for every sample of platelet count, especially when the count is lower or higher than normal by any method.
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Introduction: As most trauma patients require long‑term hospital stay and long‑term antibiotic therapy, the risk of fungal infections in such patients is steadily increasing. Early diagnosis and rapid treatment is life saving in such critically ill trauma patients. Aims: To see the distribution of various species of Candida among trauma patients and compare the accuracy, rapid identification and cost effectiveness between VITEK 2, CHROMagar and conventional methods. Settings and design: Retrospective laboratory‑based surveillance study performed over a period of 52 months (January 2009 to April 2013) at a level I trauma centre in New Delhi, India. Materials and Methods: All microbiological samples positive for Candida were processed for microbial identification using standard methods. Identification of Candida was done using chromogenic medium and by automated VITEK 2 Compact system and later confirmed using the conventional method. Time to identification in both was noted and accuracy compared with conventional method. Statistical analysis: Performed using the SPSS software for Windows (SPSS Inc. Chicago, IL, version 15.0). P values calculated using χ2 test for categorical variables. A P < 0.05 was considered significant. Results: Out of 445 yeasts isolates, Candida tropicalis (217, 49%) was the species that was maximally isolated. VITEK 2 was able to correctly identify 354 (79.5%) isolates but could not identify 48 (10.7%) isolates and wrongly identified or showed low discrimination in 43 (9.6%) isolates but CHROM agar correctly identified 381 (85.6%) isolates with 64 (14.4%) misidentification. Highest rate of misidentification was seen in C. tropicalis and C. glabrata (13, 27.1% each) by VITEK 2 and among C. albicans (9, 14%) by CHROMagar. Conclusions: Though CHROMagar gives identification at a lower cost compared with VITEK 2 and are more accurate, which is useful in low resource countries, its main drawback is the long duration taken for complete identification.
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Background: Human plasma paraoxonase1 (PON1) is an esterase catalyzing the hydrolysis of organophosphorus pesticides and other xenobiotics. The aims of this study were to develop a rapid method to determinate PON1 activity, evaluate some interference, and study the influence of storage temperature on PON1 activity assay. Methods: Measurement of PON1 activity was performed for 369 samples by measuring the hydrolysis of paraoxon using a spectrophotometric method adapted on konelab 30 . Results: The developed method facilitates the determination of PON1 activity at the rate of more than 200 samples per hour, and it is linear between 2 and 900 IU/L. Intra and inter-assay imprecision coefficients of variation were 2% and 5% respectively. PON1 activity in serum was correlated with those in heparinized plasma (r = 0.994, p < 0.001) and in plasma/EDTA (r = 0.962, p < 0.001). The mean inhibition of the PON1 activity was, by EDTA/K3, 41 ± 10 %. There was not significant PON1 activity variation after 40 days of storage at -20°C or at +4 C. There were no substantial interferences from haemoglobin, jaundice and hyperlipidemia. Conclusion: The developed method is reliable, reproducible, and suitable. It can also be performed on heparinized plasma for the determination of PON1 activity. Hence, it may be useful for assaying PON1 activity in several intoxications such as organophosphorus, sarin, and soman nerve agents.
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Objective To study an improved automated method for isolation and purification of large amounts of human pancreatic islets from large mammals,and try to create conditions for preparation and isolation of large amounts of human islets.Methods An improved automated system was used to isolate and purify panreatic islets of sogs.Under the general anaesthesia(GA) condition,the pancreas of the dogs was via in situ vascular perfused using cold HC-A solution and then removed en bloc with duodenum and spleen.Then they were placed in cold UW aolutian.Intraductal collagenase-Ⅴ and pefabloc(4 ℃) was delivered with controlled perfusion.lslets were dissociated in system of Ricordi Chamber and purified islets separated with Ficoll density gradient centrifugation under controlled temperature.Digestion time,islets remaining trapped in exocrine tissue,final islet purity,insulin and C-pep secretory activity,and islet recovery were observed.The purified islets were observed under light microscope and electronic microscope after 24 h culture.Results The digestion time rate was(25.0?6.0) min,islets remaining trapped in exocrine tissue was(9.4?2.4)%,final islet purify rate was(89.7?3.5)%,islet recovery after digestion was(17.2?3.6)?104 IEQ/pancreas.Islet recovery after purification was(8.3?2.0)?104IEQ/pancreas.Insulin and C-pep secretory activity of the purified islets and their ultrastructure after 24 h culture were ideal.Conclusions Our improved automated method and facilities for isolation of canine pancreatic islets are reliable,The morphology and function of the procured islets are excellent and they can foreseeably be used for large-scale preparation of huma islets for clinical use.