LABORATORY Flashcards
1
Q
BLOOD GLUCOSE Reference Ranges
A
70–105mg/dL
2
Q
GLYCOSYLATED HEMOGLOBIN Reference Ranges
A
Prediabetes
● 5.7 – 6.4%
● 39–46 mmol/mol
Presence of diabetes
● ≥6.5%
● ≥48 mmol/mol Target in diabetes
● <7.0%
● <53 mmol/mol
3
Q
Albumin Reference Ranges
A
3.5-5%
4
Q
Total Proteins Reference Ranges
A
6-8%
5
Q
Globulin Reference Ranges
A
1.8-3.2%
6
Q
A/G ratio Reference Ranges
A
1.5-2.4:1%
7
Q
BLOOD GLUCOSE Conversion factor
A
0.055
8
Q
Albumin, Total Proteins, Globulin Conversion factor
A
10
9
Q
BLOOD GLUCOSE Standard
A
100 mg/dL
10
Q
BLOOD GLUCOSE
BLANK, STANDARD, SAMPLE
A
Reagent: 1.0 ml
Water: 10 pL
Reagent: 1.0 ml
Standard reagent: 10 uL
Reagent: 1.0 ml
Serum: 10 uL
11
Q
Albumin
BLANK, STANDARD, SAMPLE
A
Reagent: 3.0 ml
Water: 20 uL
Reagent: 3.0 ml
Standard reagent: 20 uL
Reagent: 3.0 ml
Serum: 20 uL
12
Q
Total Proteins
BLANK, STANDARD, SAMPLE
A
Reagent: 3.0 ml
Water: 20 uL
Reagent: 3.0 ml
Standard reagent: 20 uL
Reagent: 3.0 ml
Serum: 20 uL
13
Q
BLOOD GLUCOSE Stand/Incubation
A
37 C for 5 minutes
14
Q
Albumin Stand/Incubation
A
1 min
15
Q
Total Proteins Stand/Incubation
A
5 mins at RT
16
Q
BLOOD GLUCOSE Absorbance
A
480 and 520 nm
17
Q
Albumin Absorbance
A
600 nm
18
Q
Total Proteins Absorbance
A
550 nm
19
Q
BLOOD GLUCOSE Method
A
TRINDER/GOD
20
Q
GLYCOSYLATED HEMOGLOBIN Method
A
Boronate Affinity Assay
21
Q
Albumin Method
A
Bromcresol Green (BCG) Method
22
Q
Total Proteins Method
A
Biuret, Modified
23
Q
BLOOD GLUCOSE Specimen
A
Serum, plasma, urine, CSF
24
Q
CREATININE Specimen
A
serum, plasma, urine
25
BLOOD GLUCOSE Stability
Separated and non-hemolyzed samples: 8 hours at 25C and 3 days at 2 – 8 C Glycolysis: 5 – 7% in 1 hour (5-10 mg/dL)
+ Sodium iodoacetate or sodium fluoride (NaF): 3 days at RT
26
Albumin Stability
60 mins.
27
Total Proteins Stability
60 mins.
28
BLOOD GLUCOSE Linearity
500 mg/dl
29
BUN Reference Ranges
10-20 mg/dL (1.7 – 8.3 mmol/L)
30
CREATININE Reference Ranges
Male: 0.7 – 1.2 mg/dL (62 - 105μmol/L)
Female: 0.6 – 1.1 mg/dL (53 - 97μmol/L)
31
BUA Reference Ranges
Male: 3.5 – 7.2 mg/dL (0.21 - 0.42 mmol/L)
Female: 2.6 – 6.0 mg/dL (0.15 - 0.35 mmol/L)
32
BUN Conversion factor
0.357
33
CREATININE Conversion factor
88.4
34
BUA Conversion factor
0.059
35
BUN Standard
50
36
CREATININE Standard
2
37
BUA Standard
10
38
BUN
BLANK
STANDARD
SAMPLE
Reagent: 2.0 ml
Water: 20 uL
Reagent: 2.0 ml
Standard reagent: 20 uL
Reagent: 2.0 ml
Serum: 20 uL
39
CREATININE
BLANK
STANDARD
SAMPLE
Reagent: 1.0 ml
Water: 100 uL
Reagent: 1.0 ml
Standard reagent: 100 uL
Reagent: 1.0 ml
Serum: 100 uL
40
BUA
BLANK
STANDARD
SAMPLE
Reagent: 1.0 ml
Water: 25 uL
Reagent: 1.0 ml
Standard reagent: 25 uL
Reagent: 1.0 ml
Serum: 25 uL
41
BUN Stand/Incubation
37C for 5 minutes
A1: 30 seconds at 37C A2: 60 seconds
42
CREATININE Stand/Incubation
37C for 5 minutes
A1: 60 seconds at 37C
A2: 60 seconds
43
BUA Stand/Incubation
37C for 5 minutes
44
BUN Absorbance
340 nm
45
CREATININE Absorbance
510 nm
46
BUA Absorbance
510-560nm
47
BUN Method
Coupled enzyme reaction (Urease & Glutamate DH)
48
CREATININE Method
Modified Jaffe Reaction
49
CREATININE Stability
24 hours at 2 – 8 C
Freeze samples for prolonged storage
50
BUA Stability
5 days at 4C (Uric acid in serum)
51
BUN Linearity
300 mg/dL
52
CREATININE Linearity
20 mg/dl
53
BUA Linearity
30 mg/dl
54
Plasma collected in anticoagulant containing either ammonium or fluoride salts cannot be used.
BUN
55
No interference was observed by the presence of:
CREATININE
56
Oxalate, citrate and fluoride could yield a small decrease in uric acid. The following substances interfere in very high doses; a-methyldopa, L-dopa, 3,4 dihydroxyphenylacetic acid, and gentisic acid.
BUA
57
Hemoglobin: ≤500 mg/dL
Bilirubin: ≤4.4 mg/dL
Lipids: ≤600 mg/dL
BUN
58
Hemoglobin: ≤200 mg/dL
Bilirubin: ≤7 mg/dL
Lipids: ≤600 mg/dL
CREATININE
59
Hemoglobin: <50 mg/dL
Bilirubin: <33 mg/dL
Lipids: ≤1,200 mg/dL
BUA
60
Total Cholesterol Reference Ranges
Desirable: 140 – 200 mg/dL
Borderline: 200 – 240 mg/dL
High Risk: >240 mg/dL
61
TRIGLYCERIDE Reference Ranges
Desirable: <200 mg/dL (2.26 mmol/L)
62
HDL – C Reference Ranges
Adult male: 35.3 – 79.5 mg/dL
Adult female: 42.0 – 88.0 mg/dL
63
LDL-C Reference Ranges
Desirable: <130 mg/dL Borderline for CHD: 130 – 159 mg/dL
High Risk for CHD: ≥160 mg/dL 76 – 218 mg/dL
64
Total Cholesterol Conversion factor
0.0259 (mmol/L)
65
TRIGLYCERIDE Conversion factor
0.011
66
Total Cholesterol Standard
200
67
TRIGLYCERIDE Standard
200 mg/dL
68
Total Cholesterol
BLANK
STANDARD
SAMPLE
Reagent: 1.0 ml
Water: 10wL
Reagent: 1.0 ml
Standard reagent: 10 pL
Reagent: 1.0 ml
Serum: 10 uL
69
TRIGLYCERIDE
BLANK
STANDARD
SAMPLE
Reagent: 1.0 ml
Water: 10wL
Reagent: 1.0 ml
Standard reagent: 10 pL
Reagent: 1.0 ml
Serum: 10 uL
70
HDL – C
BLANK
STANDARD
SAMPLE
Reagent A: 360 uL
Reagent B: 120 uL
Water: 4 uL
Reagent A: 360 uL
Reagent B: 120 uL
Calibrator: 4 pL
Reagent A: 360 uL
Reagent B: 120 uL
Serum: 4 uL
71
LDL-C
BLANK
STANDARD
SAMPLE
Reagent A: 360 uL
Reagent B: 120 uL
Water: 4 uL
Reagent A: 360 uL
Reagent B: 120 uL
Calibrator: 4 pL
Reagent A: 360 uL
Reagent B: 120 uL
Serum: 4 uL
72
Total Cholesterol Stand/Incubation
37C for 5 minutes
73
TRIGLYCERIDE Stand/Incubation
37C for 5 minutes
74
HDL – C Stand/Incubation
37C for 5 minutes
75
LDL-C Stand/Incubation
37C for 5 minutes
76
Total Cholesterol Absorbance
480 and 520 nm
77
TRIGLYCERIDE Absorbance
546 nm
78
HDL – C Absorbance
593 nm
79
LDL-C Absorbance
600 nm
80
Total Cholesterol Method
Cholesterol Oxidase (Trinder Method)
81
TRIGLYCERIDE Method
Trinder method
82
HDL – C Method
Immunologic method
83
LDL-C Method
Modified Trinder
84
TRIGLYCERIDE Specimen
Fasting specimen obtained (10 – 14 hours); either serum or EDTA
85
LDL-C Specimen
serum, heparinized or EDTA plasma
86
When EDTA plasma is used, the plasma value is converted to the equivalent serum value by multiplying the plasma value by 1.03.
TRIGLYCERIDE
87
TRIGLYCERIDE Stability
4C: 3 days
frozen at -20C: 2 weeks
frozen at -70C: longer periods
88
Lipemic specimens may require warming at 37C and vigorous mixing before analysis, especially if they have been frozen.
TRIGLYCERIDE
89
TRIGLYCERIDE Linearity
LDL-C Linearity 400 mg/dL
1000 mg/dl
90
Hemoglobin: ≤150 mg/dL
Bilirubin: ≤18 mg/dL
TRIGLYCERIDE
91
Hemoglobin: ≤500 mg/dL
Bilirubin (free): ≤50 mg/dL
Bilirubin (conjugated): ≤40 mg/dL
Ascorbic acid: ≤50 mg/dL
LDL-C
92
Direct Bilirubin Reference Ranges
Adults: ≤0.20 mg/dL (≤3.4 μmol/L)
93
Total Bilirubin Reference Ranges
Adults: 0.2 – 1.0 mg/dL (3.4 – 17.1 μmol/L)
Newborns: Up to 24 hrs. 2.0 – 6.0 mg dL (3.4 – 103 μmol/L)
Up to 48 hrs.: 6.0 – 10.0 mg/dL (103 – 171 μmol/L)
3 – 5 days: 4.0 – 8.0 mg/dL (68 – 137 μmol/L)
94
Direct Bilirubin REAGENT
Reagent A: 0.26M sodium chloride, 0.1Mm EDTA
Reagent B: 0.1mM EDTA, diazotized 2.4-dichloraniline 0.1mM, 0.18M hydrochloric acid
95
Total Bilirubin REAGENT
Reagent A : 0.1M hydrochloric acid, surfactant
Reagent B: 0.1M hydrochloric acid, 3.5-dichlorophenyl diazonium salt 2mM, non-reactive stabilizers
96
Direct Bilirubin
BLANK
STANDARD
SAMPLE
Reagent A: 1.0 mL
Reagent B: 250 uL
Water: 50 uL
Reagent A: 1.0 mL
Reagent B: 250 uL
Calibrator: 50 uL
Reagent A: 1.0 mL
Reagent B: 250 uL
Serum: 50 pL
97
Total Bilirubin
BLANK
STANDARD
SAMPLE
Reagent A: 360 uL
Reagent B: 120 pL
Water: 4 uL
Reagent A: 360 uL
Reagent B: 120 uL
Calibrator: 4 pL
Reagent A: 360 uL
Reagent B: 120 uL
Serum: 4 uL
98
Direct Bilirubin
Total Bilirubin
Stand/Incubation
25, 30 or 37C for 5 minutes [(Ac1) > (Ax1) > (Ac2) > (Ax2)]
99
Direct Bilirubin
Total Bilirubin
Method
Modified Jendrassik-Grof Method
100
Direct Bilirubin Absorbance
520-560 nm
101
Total Bilirubin Absorbance
490-520 nm
102
Fresh, hemolysis-free serum or heparinized plasma may be used. Carefully protect the sample from light until use. Bilirubin in sample is stable for 3 days when stored in the dark at 2-8 ̊C and 1 month at -20C Direct
Direct Bilirubin
Total Bilirubin
103
Bilirubin Linearity
13 mg/dL
104
Total Bilirubin Linearity
20 mg/dL
105
Hemoglobin: ≤50 mg/dL
Lipids: ≤500 mg/dL
Ascorbic acid: ≤35 mg/dL
Direct Bilirubin
106
Hemoglobin: ≤150 mg/dL
Lipids interfere
Total Bilirubin
107
Conjugated (direct) bilirubin reacts with diazotized 2,4-dichloroaniline in acidic solution to produce an intensely colored red diazo compound (520-560 nm). The intensity of color of this dye in solution is proportional to the concentration of direct bilirubin.
Direct Bilirubin
108
Bilirubin reacts with diazotized 3.5-dichloroaniline to produce an intensely colored diazo compound (490-520 nm). The intensity of color of this dye in solution is proportional to the concentration of total bilirubin. Free bilirubin is not soluble in aqueous media, but this reagent contains an association of surfactant and accelerators in order to provide an accurate measurement of total bilirubin.
Total Bilirubin
109
The enzyme glucose oxidase catalyzes the oxidation of glucose to gluconic acid and H2O2. The H2O2 reacts with phenol and 4-aminoantipyrine in the presence of peroxidase to form a quinoneimine dye. The intensity of the color formed (pink) is proportional to the glucose concentration and can be measured photometrically between 480 and 520 nm.
BLOOD GLUCOSE
110
The CERA-STATTM 4000 is a boronate affinity assay. The CERA-STATTM HbA1c Test Kit consists of the test device, the R1 reagent and the R2 reagent. The R1 reagent contains the agents that lyse erythrocytes and precipitate hemoglobin specifically, as well as a blue boronic acid conjugate that binds cis-diol of glycated hemoglobin.
GLYCOSYLATED HEMOGLOBIN
111
When blood is added to the R1 reagent, the erythrocytes are lysed and all hemoglobin precipitates. The boronic acid conjugate binds to the cis-diol configuration of glycated hemoglobin. An aliquot of the reaction mixture is added to the test device and all the precipitated hemoglobin, conjugatebound and unbound, remains on top of the filter. Any unbound boronate is removed with the R2 reagent. The precipitate is evaluated by measuring the blue (glycated hemoglobin) and the red (total hemoglobin) color intensity respectively with the CERA-STATTM 4000 Analyzer, the ratio between them being proportional to the percentage of glycated hemoglobin in the sample.
GLYCOSYLATED HEMOGLOBIN
112
Determination of Albumin in serum or plasma is usually based on the binding behavior of the protein with an anionic dye, bromcresol green forming a green colored complex.
Serum Albumin
113
When a solution containing proteins is rejected with a tartrate-complex cupric ions and alkali, the copper binds to the peptide bond structure of the proteins, forming a purple colored chromogen.
Total Proteins
114
The urease enzyme hydrolyzes urea in the sample to release ammonium ions, which react with 2oxoglutarate and NADH in the presence of glutamate dehydrogenase to form glutamate and NAD. The decrease in absorbance is measured at 340 nm.
BUN
115
Creatinine reacts with picric acid in an alkaline environment to form a red-colored product (creatinine picrate). The intensity of the red color is directly proportional to creatinine concentration which can be measured at 500-520 nm. The use of a surfactant and sodium tetraborate keeps interferences at minimum
CREATININE
116
Uric acid in the sample is oxidized by uricase to allantoin and hydrogen peroxide. Hydrogen peroxide reacts with ADPS and 4-aminoantipyrine in the presence of peroxidase to yield quinoneimine dye. The intensity of the dye is measured photometrically (510-560) and is directly proportional to the concentration of uric acid in the sample.
BUA
117
Cholesterol ester hydrolase hydrolyzes cholesterol esters to free cholesterol and fatty acids. Cholesterol oxidase catalyzes the oxidation of free cholesterol to cholest-4-ene-3-one and hydrogen peroxide. The H2O2 reacts with p-chlorophenol and 4-aminoantipyrine in the presence of peroxidase to form a quinoneimine dye. The intensity of color formed is proportional to the cholesterol concentration and can be measured photometrically between 480 and 520 nm.
Total Cholesterol
118
Triglycerides are hydrolyzed by lipoprotein lipase to produce glycerol and free fatty acids. The glycerol participates in a series of coupled enzymatic reactions in which glycerol kinase / glycerol phosphate oxidase are involved and H2O2 is generated. H2O2 reacts with TOPS and 4aminoantipyrine in the presence of peroxidase to form a quinoneimine dye. The intensity of color formed is proportional to the triglyceride concentration and can be measured photometrically at 546 nm
Triglycerides
119
Anti-human 3 lipoprotein antibody in Reagent A binds to lipoproteins (LDL, VLDL and chylomicrons) other than HDL. The antigen-antibody complexes formed block enzyme reactions when Reagent B is added. Cholesterol esterase and cholesterol oxidase in Reagent B react only with HDL-C. Hydrogen peroxide produced by the enzyme reactions with HDL-C yields a blue color complex upon oxidative condensation of F-DAOS [N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5dimethoxy-4-fluoroaniline, sodium salt] and 4-aminoantipyrine in the presence of peroxidase. By measuring the absorbance of the blue color complex produced, at the optimum wavelength of 593 nm, the HDL-C concentration in the sample can be calculated when compared with the absorbance of the HDL-C calibrator.
HDL – C
120
When a sample is mixed with Reagent 1, the protecting reagent binds to LDL and protects it from enzyme reactions. Cholesterol esterase (ChE) and cholesterol oxidase (ChO) react with non-LDL lipoproteins (chylomicrons, VLDL and HDL). Hydrogen peroxide produced by the enzyme reactions with non-LDL cholesterol is decomposed by catalase in Reagent 1. When Reagent 2 is added, the protecting agent is removed from LDL and catalase is inactivated. In this second process, ChE and ChO react only with LDL-C. Hydrogen peroxide produce by the enzyme reactions with LDL-C yields a color complex upon oxidase condensation with N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline (HDAOS) AND 4aminoantipyrine (4AA) in the presence of peroxidase (POD). By measuring the absorbance of the blue color complex produced at approximately 600 nm, the LDL-C concentration in the sample can be calculated when compared with the absorbance of the LDL-C calibrator.
LDL-C
