1.Effects of Peripheral Blood Different Pretreatment Methods and Preservation Time on RNA Quality.
Jia-Yi ZHANG ; Qian-Nan XU ; Xi-Ling LIU ; Cheng-Tao LI
Journal of Forensic Medicine 2021;37(6):825-831
OBJECTIVES:
To evaluate the effects of different pretreatment methods and preservation time on RNA quality of peripheral blood samples, and to optimize the preservation method of peripheral blood samples.
METHODS:
Eight pretreatment methods were used to preprocess the peripheral blood from 3 healthy unrelated individuals and the treated samples were stored at -80 ℃. Total RNA of samples was extracted using Quick-RNATM Miniprep Plus kit. DNA/RNA ShieldTM was added to peripheral blood and total RNA was extracted after preservation at -80 ℃ for 0, 5, 10, 15, 30 and 60 days, respectively. The concentration, purity and integrity of RNA were determined. Statistical analyses were performed by SPSS 22.0 software to compare the differences in RNA yield, purity and integrity among the eight pretreatment methods.
RESULTS:
In terms of purity, leukocyte pretreated with RNAlaterTM and directly cryopreservation peripheral blood showed the worst purity. The other six methods showed better purity. In terms of yield, blood cells with DNA/RNA ShieldTM came out with the highest yield, followed by peripheral blood with DNA/RNA ShieldTM. In terms of integrity, peripheral blood preserved in PAXgene Blood RNA tube method had the best integrity. Except for peripheral blood pretreated with DNA/RNA ShieldTM and blood cells pretreated with DNA/RNA shieldTM, the other five methods had statistical differences when compared to the method by keeping peripheral blood in PAXgene Blood RNA tube. The purity of RNA stored at six-time gradients ranged from 1.815 to 1.952. With the increase of storage time, RNA yield decreased from 4.516 ng to 1.039 ng, and RNA integrity decreased from 8.533 to 7.150.
CONCLUSIONS
According to the results of total RNA's yield, purity and integrity, peripheral blood pretreated with DNA/RNA ShieldTM was the best pretreatment method. After the pretreatment, samples can be preserved for up to 60 days in low temperature.
Blood Specimen Collection/methods*
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Cryopreservation
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DNA/analysis*
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Humans
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RNA
2.Effects of Two Placement Ways for Storage of Blood Bag on Biochemical Indexes of Leukodepleted Red Blood Cells.
Rui-Jun ZHANG ; Bing-Zheng DUAN ; Chun-Mei JU ; Su-Qin SUI ; Yan BAI ; Huan CAO
Journal of Experimental Hematology 2016;24(2):607-610
OBJECTIVETo investigate the effects of 2 different ways of storage bag placement on some biochemical indexes of leukodepleted red blood cells (LD-RBC) to as to ensure the efficacy and safety of clinical blood transfusion.
METHODSThe whole blood samples of 20 donors (400 ml/donor) were selected for preparating the LP-RBC, which were divided evenly into 10 bags. The 10 bags were randomly divided into 2 groups; the bags in 1 group were placed uprightly, while the bags in another group were placed horizontally. The bags of 2 groups were stored in the same conditions. One storage bag from each group was taken randomly on day 7, 14, 21, 28, 35 respectively, and then the biochemical indexes of samples were detected and analyzed.
RESULTSThe values of K(+) and LAC on day 14, the value of LDH on day 28 in the uprightly placed group were higher than those in the horizontally placed group (P < 0.05), the value of Na(+) on day 28, and the value of Glu on day 35 in the uprightly placed group were lower than those in horizontally placed group (P < 0.05), but there was no significant difference in Cl(-) level between 2 groups (P > 0.05).
CONCLUSIONThe storage bags placed by different ways during the storage show different influence on some biochemical indexes of LD-RBC in the storage period.
Blood Specimen Collection ; instrumentation ; methods ; Blood Transfusion ; Erythrocytes ; Humans ; Random Allocation
3.Effects of additives in blood collection tubes on testing the alcohol concentration in blood samples.
Journal of Forensic Medicine 2014;30(6):452-455
OBJECTIVE:
To discuss blood collection tubes with different additives and their effects on the testing results of alcohol concentration in blood samples.
METHODS:
Blood samples from 10 volunteers were collected 2 hours after drinking with seven different types of disposable vacuum blood collection tubes, including ordinary tube without anticoagulant, coagulant tube, separating gel-coagulant tube, sodium citrate (1:4) tube, sodium citrate (1:9) tube, sodium citrate (9:1) tube and EDTA-K2 tube. The alcohol concentrations in these blood samples were analyzed by headspace gas chromatography.
RESULTS:
The concentration testing results of the same blood samples in different types of tubes were different from one to another. The sequence was as follows: separating gel-coagulant tube > coagulant tube > ordi- nary tube without anticoagulant > EDTA-K2 tube> sodium citrate (1:9) tube> sodium citrate (1:4) tube, whereas the results of the same blood sample in sodium citrate (1:9) tube and sodium citrate (9:1) tube showed no obvious difference.
CONCLUSION
It is better to collect a suspicious drunk driver's blood sam- ple using a disposable vacuum blood collection tube, with the EDTA-K2 tube being preferred.
Anticoagulants
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Blood Specimen Collection/methods*
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Citrates
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Ethanol/blood*
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Humans
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Sodium Citrate
4.Application of a calling and queuing system in blood sampling in the clinical laboratory.
Da-Gan YANG ; Xi-Chao GUO ; Gen-Yun XU ; Yu CHEN
Chinese Journal of Medical Instrumentation 2008;32(2):139-141
This paper introduces the application of a calling and queuing system for blood sample collection in a large hospital in China. Besides the basic function, it has following functions. (a) A real name system: get the number according to the laboratory application form to prevent the phenomena of buying a number and an empty number. (b) Two times waiting: the patient should wait at the main hall, then at the blood sampling window so as to improve the work efficiency. (c) The flowchart for an outpatient blood testing is as following: getting the number --> waiting --> blood sampling --> getting the test information report. This system is capable of not only optimizing the work flow, but also improving the clinical environment. It shortens the patient's waiting time and raises the laboratory quality as well.
Ambulatory Care
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methods
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Ambulatory Care Information Systems
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Blood Specimen Collection
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Laboratories, Hospital
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organization & administration
5.Effectiveness of Sodium Fluoride as a Glycolysis Inhibitor on Blood Glucose Measurement: Comparison of Blood Glucose using Specimens from the Korea National Health and Nutrition Examination Survey.
Yong Wha LEE ; Young Joo CHA ; Seok Lae CHAE ; Junghan SONG ; Yeo Min YUN ; Hae Il PARK ; Moon Woo SEONG ; Dong Hee WHANG ; Hyun Soo KIM ; Jeong Ho KIM ; Bong Suk LEE ; Yoo Sung HWANG
The Korean Journal of Laboratory Medicine 2009;29(6):524-528
BACKGROUND: Accurate measurement of blood glucose concentrations is essential for defining diabetes, and the minimization of ex vivo glycolysis has been recommended. Recent guidelines advocate two kinds of methods for sample collection and processing: either the sodium fluoride (NaF) method or immediate refrigeration using a serum separation tube (SST). We investigated the difference between the two methods in measuring subsequent glucose concentrations using blood specimens from participants recruited for the fourth Korean National Health and Nutrition Examination Survey. METHODS: Paired venous blood samples were collected in an SST and a NaF tube from 1,103 men and women. SST serum was separated within 30 min, including standing for 15 min, and then refrigerated. The NaF samples were refrigerated, but not separated until immediately before analysis. We compared the blood glucose concentrations between the SST (SST glucose) and NaF (NaF glucose) methods. RESULTS: The mean SST glucose was significantly higher than NaF glucose (99.0 mg/dL vs 96.5 mg/dL, P<0.05). NaF glucose showed a negative mean bias of 2.6 mg/dL vs SST glucose but showed high correlation (R=0.9899). There was no significant correlation between the bias of blood glucose concentrations by two methods and the storage time of NaF glucose. CONCLUSIONS: The negative bias associated with the use of NaF tubes may significantly affect the prevalence of diabetes. Serum separation and refrigeration within 30 min after venous sampling is recommended over NaF method, not only to minimize the preanalytical impact on detecting diabetes but also to reduce sample volume and number of tubes.
Blood Glucose/*analysis
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Blood Specimen Collection/*methods
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Diabetes Mellitus/diagnosis
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Female
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Glycolysis/*drug effects
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Humans
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Male
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Nutrition Surveys
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Republic of Korea
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Sodium Fluoride/*pharmacology
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Specimen Handling
6.Evaluation of the Abbott Cell-Dyn Sapphire Hematology Analyzer.
Younhee PARK ; Jaewoo SONG ; Sungwook SONG ; Kyung Soon SONG ; Mee Suk AHN ; Mi Sook YANG ; Il KIM ; Jong Rak CHOI
The Korean Journal of Laboratory Medicine 2007;27(3):162-168
BACKGROUND: The performance of Cell-Dyn Sapphire (Abbott Diagnostic, USA) was compared to the Bayer Advia 2120 (Bayer Diagnostics, USA), Sysmex XE-2100 (Sysmex Corporation, Japan), and reference microscopy. METHODS: Three hundred samples for routine CBC and WBC differentials were randomly chosen for a comparison analysis. The Cell-Dyn Sapphire system was evaluated according to the linearity, imprecision, inter-instrument correlations, and white blood cell differential. RESULTS: The CBC parameters (WBC, RBC, hemoglobin and platelet) showed a significant linearity with correlation coefficients greater than 0.99 (P<0.0001). Coefficients of variation (CV) for withinrun and differential count of WBC were less than 5% except for Total CV for monocytes, eosinophils, and basophils and within-run CV for low valued eosinophils. The correlation coefficients with manual count were lower in monocytes, eosinophils, and basophils than in neutrophils and lymphocytes. The correlation with other hematology anlayzers was significant exclusive of basophils. CONCLUSIONS: These results demonstrate that the Cell-Dyn Sapphire has a good linearity, an acceptable reproducibility, a minimal carryover, and a comparable performance with the sysmex XE-2100 and Advia 2120.
Analysis of Variance
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Autoanalysis
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Blood Cell Count/*instrumentation/methods
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Blood Specimen Collection
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Diagnostic Errors
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Humans
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Reproducibility of Results
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Sensitivity and Specificity
7.Liquid Heparin Anticoagulant Produces More Negative Bias in the Determination of Ionized Magnesium than Ionized Calcium.
Cheung Soo SHIN ; Chul Ho CHANG ; Jeong Ho KIM
Yonsei Medical Journal 2006;47(2):191-195
The ionized calcium level in blood is known to be falsely decreased when self-prepared liquid heparin anticoagulant is used, due to dilution and binding effects. The effect of liquid heparin on the determination of ionized magnesium is not as well understood. We compared the effect of liquid sodium heparin on the determination of ionized calcium and magnesium in 44 clinical samples using two types of user-prepared heparin syringes which differed in the amount of residual heparin from the BD Preset(TM) reference syringe. With the type 1 syringe, the liquid heparin was expelled once or twice such that some heparin could be left in the dead space at the syringe hub, while the liquid sodium heparin was thoroughly expelled from the type 2 syringe. The ionized magnesium levels obtained with the type 1 syringe were significantly lower than the reference value (by 0.068 mmol/L) (p < 0.0001), while the value obtained with the type 2 syringe differed less from the reference, by only 0.014 mmol/L (p < 0.0001). The heparin binding effect resulted in more negative bias in ionized magnesium (-0.026 +/- 0.032 mmol/L) than in ionized calcium (-0.009 +/- 0.042 mmol/L, p < 0.0001). In conclusion, we recommend using lyophilized, calcium-balanced, heparinized syringes for the determination of ionized magnesium and ionized calcium due to the increased negative bias in ionized magnesium determinations. When user-prepared syringes are used, the thorough evacuation of heparin solution should be strictly prescribed.
Syringes
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Protein Binding
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Magnesium/*chemistry/metabolism
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Ions
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Humans
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Heparin/administration & dosage/*therapeutic use
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Calcium/*metabolism
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Blood Specimen Collection/*methods
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Blood Chemical Analysis/*methods
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Anticoagulants/therapeutic use
8.Evaluation of SD BIOLINE Chagas Ab Rapid Kit.
Mi Jung JI ; Jae Sang NOH ; Byung Ki CHO ; Young Shik CHO ; Sun Joo KIM ; Byoung Su YOON
The Korean Journal of Laboratory Medicine 2009;29(1):48-52
BACKGROUND: Chagas' disease is caused by Trypanosoma cruzi, a protozoan parasite, which is transmitted by blood-sucking bugs or through blood transfusion or organ transplantation. It is endemic in Central and South America. The objective of this study was to compare the performance of immunochromatographic SD Bioline Chagas Ab Rapid (Standard Diagnostics, Korea) with three immunochromatographic kits for the detection of antibodies to T. cruzi. METHODS: A total of 320 serum specimens (140 positive and 180 negative) from National Reference Laboratory for Chagas and Leishmaniasis (NRLCL, Honduras) were used for the evaluation of four different test kits: SD Bioline Chagas Ab Rapid, Chagas Stat-Pak Assay (Chembio Diagnositc Systems, USA), OnSite Chagas Ab Rapid test-Cassette (CTK Biotech, USA), and Trypanosoma Detect Rapid Test (InBios International, USA). The results of four kits were compared with those of NRLCL. Cross-reactivity with other parasites was also evaluated. RESULTS: Compared with the results of NRLCL, sensitivity and specificity were 99.3% and 100% for both of SD and Chembio kits, 97.2% and 100% for InBios kit, and 97.9% and 98.8% for CTK kit. None of other parasites showed cross-reactivity. CONCLUSIONS: SD Bioline Chagas Ab Rapid kit showed test results highly correlating with those of National Reference Laboratory for Chagas and Leishmaniasis. It can be used for a rapid detection of Chagas' disease in endemic region and monitoring the disease among overseas travelers in Korea.
Animals
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Antibodies, Protozoan/*blood/immunology
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Blood Specimen Collection
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Chagas Disease/*diagnosis/immunology/parasitology
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Chromatography/methods
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Humans
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*Reagent Kits, Diagnostic
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Sensitivity and Specificity
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Trypanosoma cruzi/*immunology
9.Evaluation of PIMA point-of-care CD4 analyzer in Yunnan, China.
Jun LIANG ; Song DUAN ; Yan-Ling MA ; Ji-Bao WANG ; Ying-Zhen SU ; Hui ZHANG ; Chin-Yih OU ; Ling HAO ; Ming-Shan QI ; Marc BULTERYS ; Larry WESTERMAN ; Yan JIANG ; Yao XIAO
Chinese Medical Journal 2015;128(7):890-895
BACKGROUNDCD4 count is used to determine antiretroviral therapy (ART) eligibility. In China, flow cytometers are mostly located in urban areas with limited access by patients residing in remote areas. In an attempt to address this issue, we conducted a study to validate the performance of Alere PIMA point-of-care CD4 analyzer.
METHODSVenous and finger-prick blood specimens were collected from HIV-positive participants from two voluntary counseling and testing sites in Yunnan Province. Both venous and finger-prick blood specimens were tested with the PIMA analyzer. Venous blood specimens tested with the Becton Dickinson FACSCalibur were used as a reference.
RESULTSVenous specimens from 396 and finger-prick specimens from 387 persons were available for analysis. CD4 counts by PIMA correlated well with those from FACSCalibur with an R2 of 0.91 for venous blood and 0.81 for finger-prick blood. Compared to FACSCalibur, the PIMA analyzer yielded lower counts with a mean bias of - 47.0 cells/μl (limit of agreement, [LOA]: -204-110 cells/μl) for venous blood and -71.0 cells/μl (LOA: -295-153 cells/μl) for finger-prick blood. For a CD4 threshold of 350 cells/μl, the positive predictive value (PPV) of PIMA was 84.2% and 75.7% and the negative predictive value (NPV) was 97.6% and 95.8% for venous and finger-prick blood, respectively. For an ART threshold of 500 cells/μl, the corresponding PPV was 90.3% and 84.0% and NPV was 94.3% and 93.4%, respectively.
CONCLUSIONSCD4 counting using venous blood with PIMA analyzers is a feasible alternative to a large flow cytometer to determine ART eligibility.
Adolescent ; Adult ; Aged ; Biological Assay ; methods ; Blood Specimen Collection ; CD4 Lymphocyte Count ; methods ; Child ; China ; Female ; HIV Infections ; diagnosis ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult
10.A Comparison of the Rates of Hemolysis and Repeated Blood Sampling using Syringe needles versus Vacuum tube needles in the Emergency Department.
Young Hee SUNG ; Moon Sook HWANG ; Jee Hyang LEE ; Hyung Doo PARK ; Kwang Hyun RYU ; Myung Sook CHO ; Young Hee YI ; S SONG
Journal of Korean Academy of Nursing 2012;42(3):443-451
PURPOSE: This study was done to compare the rates of hemolysis and repeated sampling in blood samples obtained by a syringe needle versus a vacuum tube needle. METHODS: A randomized, prospective study was used to evaluate the differences between the two blood sampling methods. The study group consisted of patients seen in the emergency department (ED) for blood sampling to determine electrolyte level. ED patients were randomly assigned to either the syringe group or the vacuum tube group. All blood samples were collected by experienced ED nurses and hemolysis was determined by experienced laboratory technologists. Data were analyzed using Fisher's exact test and binary logistic regression. RESULTS: One hundred forty-five valid samples were collected (74 in the syringe group versus 71 in the vacuum tube group). 5 of 74 (6.8%) blood samples in the syringe group and 8 of 71 (11.3%) in the vacuum tube group hemolyzed. Repeated blood sampling occurred for 2 of 74 (2.7%) and 3 of 71 (4.2%) in each group respectively. There were no significant differences in rates of hemolysis and repeated sampling between two groups (B=1.97, p=.204; B=2.36, p=.345). CONCLUSION: Venipuncture with syringe needles can be recommended for ED nurses to obtain blood samples.
Adult
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Aged
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Blood Specimen Collection/instrumentation/*methods
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Emergency Service, Hospital
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Female
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*Hemolysis
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Humans
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Logistic Models
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Male
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Middle Aged
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Phlebotomy
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Prospective Studies
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Questionnaires
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Syringes