TEST 1 - Telenet

advertisement

1060

1

QUALITY AND AUTOMATION IN THE DETERMINATION OF THE ERYTHROCYTE SEDIMENTATION RATE

by Paolo Galiano

2

In the last 5 years many scientific works on the TEST1 system have been published by different authors, many of them Italian, but others from The Netherlands, New Zealand, Korea, Spain.

The state of the art of these articles provides a view of the new state of the art of the Erythrocyte Sedimentation Rate in the world.

3

SCIENTIFIC WORKS AND PUBLICATIONS

1. M. Plebani, S. De Toni, M.C. Sanzari, E. Stockreiter, D. Bernardi (Dept. of Laboratory Medicine, University-Hospital, Padova, Italy), “The TEST 1 Automated System: A New Method for Measuring for Erythrocyte Sedimentation Rate”,

American Journal of Clinical Pathology

, 1998, 110:334-340.

2. N. Cirilli, Z. Abu Asy, N.

Giacchè, F. Bordicchia, S. Paolucci, M. Tocchini (Dept. of Laboratory Medicine, G. Salesi Hospital, Ancona, Italy), “TEST1: Un Nuovo Metodo per la Determinazione della VES”,

Biochimica Clinica

, Vol. 22, N. 5-6, 1998, p. 339.

3. M. Plebani, S. De Toni, M.C. Sanzari, E. Stockreiter, D. Bernardi, F. Floriani (Dept. of Laboratory Medicine, University-Hospital, Padova, Italy), “Il Sistema Automatizzato TEST1: un Nuovo Metodo per la Determinazione della Velocità di Eritrosedimentazione”,

Medicina di Laboratorio

, Vol. 6, N. 2, June 1998, pp. 166-172.

4. G. Soffiati (Clinical Chemistry and Hematology Laboratory, San Bortolo Hospital, Vicenza, Italy), “Nuovo Metodo per la Determinazione della Velocità di Eritrosedimentazione (VES)”, August 1998, private communication.

5. L. Germagnoli, S. Lopez-Silva, M. Murone, S. Vazzana, L. Grassini, L. Calloni, V. Gioia (Dept. of Laboratory Medicine, Scientific Institute San Raffaele, Milan, Italy), “Evaluation of the Automatic System TEST1 ™ for Measurement of the Erythrocyte Sedimentation Rate (ESR)”, issued as scientific poster on

Clinical Chemistry

, July 1999.

4

SCIENTIFIC WORKS AND PUBLICATIONS

6. K. S. Shin, J.S. Kim, B.R. Son (Dept. of Clinical Pathology, College for Medicine Chungbuk National University, Cheongju, Korea): “Evaluation of the TEST 1 for Measuring Erythrocyte Sedimentation Rate”

Journal of Clinical Pathology and Quality Control

, Vol. 21, No. 1, 1999, pp. 223-228.

7. John Robert "Erythrocyte Sedimentation. A New Solution to an Old Problem", (Hitech Pathology, Melbourne, Australia) issued on the official publication of the

New Zealand Institute of Medical Laboratory Science and Australian Institute of Medical

Scientists,

South Pacific Congress, Christchurch, New Zealand, 23-27 August 1999, sponsored by Dade Behring Diagnostics.

8.

C. Gasparoli, D. Pulè, A. Fusco (Dept. of Laboratory Medicine, Istituto Dermopatico Immacolata – IRCCS, Rome), “Sostituzione di un Metodo Tradizionale per la Misurazione delle VES con il Nuovo Metodo Automatico TEST1”, October 1999, private communication.

9.

K. Taylor (Canterbury Health Laboratories, Christchurch, New Zealand), “TEST1 Evaluation Report”, December 1999, private communication.

10.N. de Jonge, I. Sewkaransing, J. Slinger, J.J.M. Rijsdijk (Dept. Clinical Chemistry, Leyenburg Hospital, The Netherlands), “Erythrocyte Sedimentation Rate by Test-1 Analyzer”,

Clinical Chemistry,

June 2000, 46: 881-882.

5

SCIENTIFIC WORKS AND PUBLICATIONS

11. M. Plebani, E. Piva, M.C. Sanzari, G. Servidio (Dept. of Laboratory Medicine, University Hospital, Padova, Italy), “Length of Sedimentation Reaction in Undiluted Blood (Erythrocyte Sedimentation Rate): Variations with Sex and Age and Reference Limits”,

Clinical Chemistry and Laboratory Medicine

, May 2001, 39: 451-454.

12.D. Giavarina, S. Capuzzo, M. Carta, F. Cauduro, G. Soffiati (Clin. Chem. & Hematol. Lab., San Bortolo Hospital, Vicenza, Italy), “Internal Quality Control for Erythrocyte Sedimentation Rate (ESR) measured by TEST 1 Analyzer”,

Clinical Chemistry,

June 2001, 47: 162.

13.

D. Spedding, D. Smith (Dade Behring Diagnostics, New Zealand), “Evaluation of Agreement between the TEST1 and Starrsed Automated ESR Analysers”, November 2001, private communication.

14.M. Plebani, E. Piva (Dept. of Laboratory Medicine, University-Hospital, Padova, Italy), “Erythrocyte Sedimentation Rate. Use of Fresh Blood for Quality Control”,

American Journal of Clinical Pathology

, 2002, 117:621-626.

15.E. Heverin (Galway Mayo Institute of Technology, Ireland), “Comparison of the Westergren method versus the TEST1 technique for determining the Erythrocyte Sedimentation Rate”, May 2002, private communication.

6

SCIENTIFIC WORKS AND PUBLICATIONS

16.P. Napoli, B. Montaruli, S. Plateroti, A. Martini, A. Sacchi, A. Toja. M. Saitta (Analysis Lab, CIOV, Evangelico Valdese Hospital, Turin, Italy), Controllo di “Sistema Automatizzato TEST1: Qualità Interno per la Determinazione della VES”,

Biochimica Clinica

, Vol. 26, N. 3, 2002, p. 215.

17.B.H. Lee, J. Choi, M.S. Gee, K.K. Lee, H. Park (Dept. of Laboratory Medicine, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, Korea), “Basic Evaluation and Reference Range Assessment of TEST1 for the Automated Erythrocyte Sedimentatioon Rate”,

Journal of Clinical Pathology and Quality Control

, Vol. 24, No. 1, 2002.

18.D. Giavarina, S. Capuzzo, F. Cauduro, M. Carta, G. Soffiati (Clin. Chem. & Hematol.

Lab., San Bortolo Hospital, Vicenza, Italy), Sedimentation Rate Measured Test 1 “Internal Quality Control for Erythrocyte Analyzer”,

Clinical Laboratory

2002, 48: 459 462.

19.Romero A., Victoria", Muñoz M., Ramirez G. (Dept. of Haematology, H.C.U. "Virgen de la Málaga & *GIEMSA, School of Medicine, University of Málaga, Spain), "Determination of the Length of Sedimentation Reaction in Blood: a Comparison of the Test1 ESR System with the ICSH Reference Method and the Sedisystem",

Chemistry and Laboratory Medicine

2003, 41 (2).

Clinical

7

• • • • •

SCIENTIFIC WORKS AND PUBLICATIONS

20. M. Plebani (Dept. of Laboratory Medicine, University Hospital, Padova, Italy), “Erythrocyte Sedimentation Rate: Innovative Techniques for an Obsolete Test?”,

Clinical Chemistry and Laboratory Medicine

, 2003, 41 (2): 115-116.

21. Nicoli M., Lanzoni E., Massocco A., Franceschini C.* (Laboratory of Clinical Chemistry and Haematological Analysis, Ospedale Civile Maggiore, Verona, Italy & *Dasit, Cornaredo, Italy), “Integrated Haematology and Coagulation Laboratory”, Poster,

Euromedlab Congress

, Barcelona, Spain, 1-5 June 2003.

22. P. Galiano, “Quality and Automation in the Determination of the Erythrocyte Sedimentation Rate, Symposium 046,

22nd World Congress of Pathology & Laboratory Medicine

, 30 August-2 September 2003, Busan, Korea.

23. J.M. Jou, “La VSG: còmo, cuàndo y para qué puede ser ùtil”, (Hospital Clinic University of Barcelona),

AEHH/SETH Hematology Society National Congress

, 23-25 October 2003, Santiago de Compostela, Spain. 24. B. Olivera Alonso, M. Sirvent Monerris, M.T. Rotella Belda, V. Ballenilla Antón, García Vidal (M Laboratorio Hospital San Vicente y Area Sanitaria 18. Alicante, Spain), “Cambio De Método Para La Determinación De V.S.G.: Repercusiones Sobre La Fase Preanalítica”,

Generalitat Valenciana - Conselleria De Sanitat

(for Valencia Government – MOH), Spain 2004.

8

Heverin from Galway-Mayo Institute of Technology, in describing the reproducibility of the TEST1, verifies a very important point, “temperature”. To perform a reliable internal quality control it is absolutely needed to keep the samples into the refrigerator and to allow them to reach room temperature the day after before testing them again. I remind you that from NCCLS 4 th edition, approved May 2001, ESR cannot be calibrated.

9

K3EDTA @ 24hrs 4°C, K2EDTA @ 24hrs 4°C

r=0.974

From E. Heverin (Galway Mayo Institute of Technology, Ireland), “Comparison of the Westergren method vs the TEST1 technique for determining the Erythrocyte Sedimentation Rate”, May 2002 10

From E. Heverin (Galway Mayo Institute of Technology, Ireland), “Comparison of the Westergren method vs the TEST1 technique for determining the Erythrocyte Sedimentation Rate”, May 2002

Test 1 ESR results at 4hrs versus 48hrs at 4 °C K3EDTA @ 48hrs 4°C, K2EDTA @ 48hrs 4°C

r=0.964

11

From E. Heverin (Galway Mayo Institute of Technology, Ireland), “Comparison of the Westergren method vs the TEST1 technique for determining the Erythrocyte Sedimentation Rate”, May 2002 12

From E. Heverin (Galway Mayo Institute of Technology, Ireland), “Comparison of the Westergren method vs the TEST1 technique for determining the Erythrocyte Sedimentation Rate”, May 2002 13

Now let me make a brief list of the reference subjects described by the National Committee for Clinical Laboratory Standards. All the topics belong to sodium citrate collected samples.

14

15

16

17

18

19

20

21

22

23

From NCCLS, 4rd ed., Vol. 20, No. 27, December 2000

Acceptable values comparing EDTA with citrate collected samples

EDTA Fixed 20 43 20 21 22 23 24 25 26 27 28 36 37 38 39 40 41 42 43 s a m e s a m p l e

5-17

6-17 6-18 6-19 7-19 7-20 8-21 8-21 9-22 13-29 13-30 14-30 14-31 15-32 15-33 16-34 17-35 Citrate Variables 5-17 17-35

24

Just from one of the points you can read from the original document, I use page 8 to see the limits of all citrate methods and instruments using this type of preservant (sodium citrate). TEST1 has a big advantage: it works with EDTA,considered to be the best blood preservant.

In fact, if you see this slide, the variation of citrate is intrinsic to the list shown above offering an acceptable range, but variable against a fixed value offered by EDTA samples.

25

26

27

28

29

SAMPLE

THE CLASSICAL ESR

ESR 1h ESR – HEMATOCRIT 20% Area requested for well mixing Agglomerin Normal Pathological t=0 t=1h WESTERGREN 35% Normal Hematocrit range t=12 h or Centrifugation

30

ESR is a phyisical phenomenon circumscribed on time From NCCLS: recognised curve is a sigmoid type helped by the presence of specific proteins called agglomerins Without these agglomerins there is no sedimentation

31

ESR values increase

• •

Pathologies

– leukaemia, rheumatic polimialgy, rheumatoid arthritis, thromboflebitis, subacute endocarditis, pneumoniae, colangitis, osteomyelitis, SLE, pyelonephritis, post cardiosurgery, acute hearth stroke, lung stroke, linphoma, rheumatic fever, viral hepatitis, glomerular nephritis, infectious mononucleosis, breast cancer, lung cancer, hepatic metastatis, ipernephroma, uremia, acute meningithis, ictus, sarcoidosis, pelvic infections (gonococcus and others), tubercolosis, Crohn’s disease, burns, bone fracture, anemia, arthrosis, gout, cholecystitis, typhus, mumps, acute allergy, ulcerative colitis, pregnancy, postpartum, menstruation, Cytomegalovirus infection, Toxoplasmosis, Rickettsiosis, fever, appendicitis.

Drugs

– contraceptive , A Vitamin – others: B Hepatitis vaccination 32

32

ESR values decrease

33

Pathologies

– dehydration, hepatic necrosis, Thrichinosis, polycythemia, fibrinogenopenia and iperfibrinogenemia, cachexy, anticoagulants •

Drugs

– salicylates, cortisone, ACTH, Cyclophosphamide (chemotherapeutic agent), chinin, oxalate 33

Westergren method and use of Sodium Citrate show

: (

from NCCLS Vol.20 No.27 H2-A4

) - errors in the dilution step - poor temperature control - problems connected to vibrations,verticality,diameter of pipets - test must be run within 4 hours from blood collection - PCV adjustment (<0.35) - No control available - plastic tubes may interfere with red cells surface charges due to the plastic material 34

VACUTAINER TUBE IN CITRATE

Errors in the dilution phase Correct Level Exceeding level of blood (fresh tubes) NO RATIO RESPECTED insufficient level of blood (old tubes) NO RATIO RESPECTED NO OK

Correct level of blood

Dilution ratio 1:4 RATIO RESPECTED

NO

35

THE IMPORTANCE OF HEMATOCRIT

DIFFERENT RESULTS OF THE SAME SAMPLE DILUTED WITH AUTOLOGOUS PLASMA

140 120 100 80 60 40 20 0 0 Hct of 26 20 40 60 80 100 120 140 160 180 200 TIME (minutes) Hct of 28 Hct of 34 Hct of 45

Sedimentation as a function of time with different Hct values artificially obtained adding autologous plasma

(Blood

, Vol. 70, No 5, Nov. 1987: pp 1572-1576).

36

LOW INFLUENCE OF HEMATOCRIT ON TEST 1

From

Thomas L.Fabry, Mechanism of Erythrocyte Aggregation and Sedimentation, Blood, Vol. 70, No. 5 (Nov.), 1987: pp 1572-1576.

Fabry’s Formula to adjust the value of ESR

Example of adjustment for a sample with hematocrit 30.3

- Correct value = 69.2

- WG non corrected value = 114 - TEST 1 =

67 Example of adjustment for a sample with hematocrit 26.3

- Correct value = 66.3

- WG non corrected value = 127 - TEST 1 =

67

37

LOW HEMATOCRIT

The instrument indicates with an asterics (*), near the result, the samples with a low hematocrit value.

In the example reported the hematocrit value is low 38

Moreover, the commercial quality controls offered and pushed by our competitors, as you can see again, show the great variations of these values, improperly offered under this name: ranges and limits suggested in the figure show how low level is indicated from 1 to14 and high level from 15 to 55.

39

40

41

TEST1 offers much closer reproducibility as many authors have described using internal samples of their labs, tested 24 and 48 hours later. The correlations you see on the data are comparable to normal clinical chemistry controls and, finally, our statistical quality control gives you a method based on statistical data of your population. The cumulative data of any day (white circles) can be compared with the black one, that are the statistical data collected by the instrument during a month and these collected data can be considered as a standard when you have in the black circle values at least from 500-1000 results. The cumulative black circle maintains a revolving number of 6000 samples as a standard.

42

TEST 1 REPRODUCIBILITY DURING THE SAME DAY Samples collected on January 28th and processed on January 29th

29/01/2003

12.07.am

29/01/2003

12.18am

29/01/2003

12.28am

29/01/2003

15.38 pm

ESR

2 38 76 14 2 15 42 7 26 16 11 52 19 59 22 3 41 79 13 3 16 44 8 25 16 11 50 17 57 22 4 44 79 13 3 16 40 7 22 16 10 47 20 55 23 2 42 74 15 4 14 41 9 23 17 11 45 18 58 24

R = 0,99 in the correlation between column B column C =

R2 0,9944

R = 0,99 in the correlation between column B column D =

R2 0,9831

R = 0,99 in the correlation between column B column E =

R2 0,988 Samples were properly stored in the refrigerator (4°C) and let them get room temperature taking them out of the refrigerator 30-40 minutes before running the tests performed with Test 1 Instrument. Samples are properly mixed in Test 1, thus aggregated cells are re-suspended in the plasma and ESR is consequently correctly measured.

alive during the training! The red column is the reference towards which the correlations of columns C, D and E have been calculated.

CORRELATION CHART TEST 1 REPRODUCIBILITY DURING THE SAME DAY

90 y = 0,9798x + 0,4054 Samples collected on January 28th 80 R 2 = 0,9831 y = 1,0066x + 0,0901 R 2 = 0,9944 70

ESR

60 50 40 30 20 10

29/01/2003 29/01/2003

12.07.am

12.18am y = 0,9446x + 1,2142 2 R 2 = 0,988 76 41 79 3 14 2 13 3 15 42 7 26 16 11 52 19 59 22 16 44 8 25 16 11 50 17 57 22

29/01/2003

12.28am

4 44 79 13 3 16 40 7 22 16 10 47 20 55 23

29/01/2003

15.38 pm 2 42 74 15 4 14 41 9 23 17 11 45 18 58 24 0 0 20 40 60

fig 1

80 R = 0.99

44

REPRODUCIBILITY AFTER 48 HOURS Samples collected on 28/1/2003 and processed on 30/1/2003

SAMPLES HAVE BEEN STORED AT +4°C DURING THE TIME LAPSE BETWEEN 29 AND 30 JANUARY ESR 29/01/2003

12.07am

30/01/2003

9.21am

2 38 76 14 2 15 42 7 26 16 11 52 19 59 22 2 36 77 11 2 12 35 8 22 14 9 43

20

56 22

30/01/2003

9.32am

R = 0.99 correlation between columns B- columns C R = 0.99 correlation between columns B- columns D

R2 0,9832 R2 0,9844 2 41 75 12 2 12 39 8 20 15 10 44

20

55 22 The following day we took out from the refrigerator the 15 samples tested the day before and let them get room temperature for about 30-40 minutes. Then we put samples into Test 1 and the working session started. After 3 minutes of mixing cycle the measurement of ESR begun.

The results of two running sessions are reported on columns C and D. Column B has been copied and pasted from the previous Excel sheet in order to have an immediate comparison among reproducibility of Test 1 and good quality of results.

45

CORRELATION CHART

90 80

ESR

70 60 50 40 30 20 10 0 0

29/01/2003

12.07am

y = 0,9557x - 0,9483 R 2 = 0,9832 2 38 R 2 10 = 0,9844 20 14 2 15 42 7 26 16 11 52 19 59 22 30

30/01/2003

9.21am

40 2 36 77 11 2 12 35 8 22 14 9 43

20

56 22 50

30/01/2003

9.32am

60 2 41 75 12 2 12 39 8 20 15 10 44

20

55 22 70 80 R = 0.99

46

I specified that the value of our control is to be interpreted upon the reproducibility of the correlation value R=0,99 and this is a real reproducibility as a clinical chemistry control, not based on the possible variation of a sample that may also give not so reproducible results the day after. This is not so frequent but if it occurred you would be able to give the correct answer: it is the reproducibility R that creates the stability of control.

Moreover, I graphically represented the normal, pathological and myeloma conditions to show the time and curve typology of the myeloma-ESR case. I have intentionally begun a discussion in which, as a first topic, I have reminded the audience that the representation of a myeloma-ESR curve is not even reported by the NCCLS, as this organisation describes only a Sigmoid-like curve to represent a physical condition, in this case the ESR testing, and not a curve that remains plain for many minutes and then precipitates.

47

Stability Study

Storage of blood sample at 4 °C for up 24 hours caused a decrease in ESR values obtained with the TEST1: the mean difference was 2.86 ( 95% CI, 2.41-3.31) and 2.28 (95% CI, 1.90-2.65) respectively for two different analyzers with the same samples (n=1.140). The decreases were of 9% and 11% .

48

STATISTICAL QUALITY CONTROL DATA REPORT

For Range 2-120 mm/h and 2-30 mm/h Print out date Scale of values Value of cumulative average  (black circle) Value of daily average O (white circle) STD S. cumulative standard deviation STD D. daily standard deviation N.B. The values printed are only examples not to be considered as absolute but only indicative.

 (black circle ) = 6000 data of cumulative values are equivalent to your statistical population, which can be considerd a Standard 49

STATISTICAL QUALITY CONTROL DATA REPORT

For control of population to obtain internal normal range 50

51

STABILITY OF VACUTAINER TUBE IN EDTA

THE EDTA COLLECTED SAMPLE REFRIGERATED AT +4 °C CAN BE TESTED EVEN AFTER 24 HOURS FROM BLOOD COLLECTION

From

Clinical Chemistry

, Vol. 47, No. 6, Supplement 2001, p. 162 -

Internal quality control for erythrocyte sedimentation rate (ESR) measured by TEST 1 analyzer

by D. Giavarina, S. Capuzzo, M. Carta, F. Caoduro, G. Soffiati,

Clin. Chem. & Hematol Lab - San Bortolo Hospital: Vicenza, Italy

52

Technical advantages

SAVE of timemoney SAVE of waste money

(expired tube – tube volume)

SAVE of stock money

53

Quality improvements The unique method for Lab Accreditation

54

55

New software benefits for LAB ACCREDITATION The unique characteristics of ALIFAX ESR analyzers provides a Statistical Internal Quality Control useful to the certification of the lab.

The new ISO9001/UNI EN ISO 9001 – Ed. 2000 certification is used for the validation of the lab test in which the lab instrumentation, controls and, in general, the analysis system can have the recognition of a value and its maximum validation to guarantee operation functionality and reliability of the results.

• •

TEST1 TH new software offers: Control of the functionality of reading sensors for each washing Data control by a statistical quality system based on the control validity of

• • • • •

the collected data of a population

Control of the daily value relative to a population of 6000 memorized data in the instrument Control of the single result scanned 1000 times by our system for each analysis Control of sample reproducibility on samples tested the day after Control of data reproducibility at R value equivalent to 0.98-0.99

Control for samples improper to the test below the normal hematocrit values < 20 Control of data transmission to LIS.

55

TECHNICAL CHARACTERISTICS

• Autodiagnostics • Internal or external bar code reader • Interfacing through RS232 • QUERY HOST Software for PC LAB management • No dedicated tubes • Compatible with cell counter racks (Bayer, Beckman Coulter, Sysmex, Abbott, ABX, etc.) • Capacity of 60 samples random batch access • 180 samples/hr • Automatic sample mixing • Blood collection according to the ICSH recommendations • Flow control to check of eventual clots • Minimum Blood Volume > 1 ml • Working volume 150 µL of blood 56

TECHNICAL CHARACTERISTICS

• Cell measure volume 1 µL • Thermostation 37°C • First result after 3 minutes and 20 seconds • Low influence of hematocrit < 35 HCT • No carryover • Autowashing • Safe-control closed cycle system • Automatic reading and print out of the results • Totally safe waste volume tank • Waste production reduced (Test 1=3 L, others=1.000L) • Waste disposal costs reduced • Statistical internal quality control of population for normal values • Safety check card • Electronic calibration for alignment of results 57

What is

TEST 1

• The only instrument in the world which can

MEASURE

the ESR values • Remember that all other instruments on the market are simply READERS 58

• Test 1 measures the kinetics of blood sedimentation phenomenon • all the other instruments simply read the final result of this phenomenon, that is why they need more time to give results 59

TEST1

ESR

TELEMETRY

60

CAPACITY OF SEDIMENTATION AND AGGREGATION

TEST 1 studies the sedimentation and aggregation capacity of the blood red cells via optical density Every sample is read 1000 times in 20 seconds verifying the aggregation and sedimentation capacity of the blood red cells Light beam before Aggregation starting t = 0 Aggregation after 1000 scans t = 20 sec.

Light beam after 61

0 Lag phase Normal Status 0

+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +

Time (minutes)

+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +

60 62

Inflammation Status 0 Lag phase 20 seconds Precipitation phase Packaging phase 70 0

+ + + + + + + + + + + + + + + + + + + + + +

Time (minutes)

+ + + + + + + + + + + + + + -

Agglomerine

+ + + + + + + + + + + + + + + + +

60 63

0

Myeloma

Example of collapsed sample for ESR

Lag phase Precipitation phase 70 0

+ + + + + + + + + + + + + + + + + +

Packaging phase Time (minutes)

+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +

60 64

Example of a collapsed sample From 200 mm to the ratio in percentage 100 200 mm 130 ESR 70 Corpuscular Part 100 mm 65 ESR 35 Hematocrit or PCV Westergren pipet 65

Myeloma Example of collapsed sample for ESR

0 Lag phase 70 0

200 mm + + + + + + + + + + + + + + + +

Time (minutes) Precipitation phase Packaging phase 60

+ + + + + + + + + + +

130 ESR

+ + + + + + + + + + + + + + + +

Westergren pipet 70 Corpuscular Part 100 mm 65 ESR

+ + + + + + + + + + +

35 Hematocrit or PCV 66

These blood samples without “ Agglomerins” do not express a ESR value with a “sigmoid” tract, but collapse to the level of the Hematocrit. Final point of reading. 67

CALCULATION METHODOLOGY OF RESULTS

The mathematical algorithm converts the results from optical density to ERYTHROCYTE SEDIMENTATION RATE of the microvolume analyzed

from sample optical density to mm/hr Westergren

68

Test-1 ESR expresses the vitality of aggregation also 12, 24, 48 hr after collection, according to the sigmoid curve of the classic graph, that is the only curve described and represented. 69

PATENT The world patent of Test-1 concerns the mathematic algorithm that expresses the same sigmoid function of the ESR 70

A synthetic control with no active electrical charge and with no agglomerins that induce and activate the sedimentation process is not recognized by Test-1 as these controls give no signals of vitality because they are not part of the “sigmoid sedimentation”, but a mixture of water – sand and not a measure of control. 71

• The Test 1 technology is PATENTED

72

73

74

30 µL of blood suitable for pediatric use

75

• Micro Test 1 can work with fresh blood , as soon as it has been collected, without need of anticoagulant or preservatives

76

The only instrument that works without preservant

77

Methodology advantages and characteristics

78

METHODOLOGY ADVANTAGES

Quality Control

• Urgent diagnosis can be fulfilled • Pediatric use • No anticoagulant use- fresh blood for MicroTest1 • Test 1

Measures

ESR,

it is not a

Reader Test 1 is an Analyzer !!!!

79

METHODOLOGY ADVANTAGES

• Capillary • Microvolume 150 or 30 microliters (T1 or MicroT1) • Photometric-kinetic • Scanning rate -total scans 1000 for sample in 20 sec.

• Not influenced from hematocrit • Rapidity of response correlated to Westergren • Reproducibility • Stability during time (EDTA-48 hours against Na Citrate 4 hours) 80

During 20 sec. the sample is scanned 1000 times

Ident. 05483311 ESR = 118 mm/hr Ident. 05725053 ESR = 26 mm/hr Ident. 05725044 ESR = 2 mm/hr ==> Time sec.

==> Time sec.

==> Time sec.

81

METHODOLOGY CHARACTERISTICS

1 It starts from 0 time to 20 sec. following and measuring the evolution of the sed rate curves.

2 It measures the optical density related to the concentration of the erythrocytes/aggregates present at the moment of the analysis.

3 Kinetics following the evolution of the curves with a frequency of 50 measures per second.

4 The capillary system simulates a invivo situation and guarantees minimal optical paths in blood subject to sedimentation which enable the detection of small variations.

82

CORRELATION TEST 1-WESTERGREN

From Clinical Chemistry and Laboratory Medicine, February 2003, 41(2)

Romero A., Muñoz M., Ramirez G., Dept. of Haematology, H.C.U. "Virgen de la Victoria", of Medicine, University of Málaga & GIEMSA, School Málaga, Spain, “Determination of the Length of Sedimentation Reaction in Blood: a Comparison of the Test1 ESR System with the ICSH Reference Method and the Sedisystem ” .

“… the correlation coefficient was 0.98…”

83

COMPARATIVE SCHEDULE WEIGHT OF WASTE DISPOSAL COMPANY

DIESSE BD

TYPE OF PRODUCED WASTE

DEDICATED TUBE + BLOOD TUBE WITH BLOOD

WASTE PRODUCED FOR EACH TESTING

gr. 12 gr. 10

WASTE PRODUCED FOR 20.000 TESTING IN KG.

Kg. 240 Kg. 200

ALIFAX

ON A COLLECTION TANK CAPACITY 2.000 TESTING gr. 0,25

Kg. 5

84

Slides from Customers

85

Imprecision of TEST 1 and Westergren

TEST 1 Westergren

ESR value

13.1

8.0

TEST 1 Westergren 29 25 TEST 1 Westergren 54.7

57.1

Range

12-14 8.0-10 27-32 21-30 51-58 51-60

CV%

4.86

9.68

5.30

8.45

3.37

5.04

86

ADVANTAGES OF EDTA AS AN ANTICOAGULANT

• •

It preserves the red blood morphology.

Does not interfere with mechanisms that lead to erythrocyte sedimentation.

Increases specimen stability.

Does not incur problems related to sample dilution with sodium citrate.

87

ADVANTAGES OF EDTA AS AN ANTICOAGULANT

Specimens anticoagulated with EDTA:

Are suitable for internal quality control programs.

Allow the creation of a unique workstation for measuring ESR and performing other hematological tests (erythrocyte, leukocyte, reticulocyte counts and differential analysis in a single specimen).

88

Reference limits, 2.5

th and 97.5

th pervcentiles and their 95% confidence intervals (CI) for SRB in undiluted EDTA-anticoagulant blood. Variation with age and sex

Age (year) n Sex 2.5 percentiles and 95%CI 97.5 percentiles and 95%CI

0-14 15-50 15-50 51-70 51-70 >70 80 190 150 120 130 170 W and M Women Men Women Men W and M 2 (2-2) 2 (2-2) 2 (2-2) 2 (2-3) 2 (2-2) 3 (3-3) 34 (26-41) 37 (36-39) 28 (20-30) 39 (38-45) 37 (31-44) 46 (45-55)

Clin Chem Lab Med 2001;39(5):451-4.

89

TEST 1 TH

150 µL of blood

90

ROLLER

91

TEST 1 TH RACK Any kind of vacutainer tube available on the market can be used to perform the ESR, e.g.:

TEST 1 TH works by loading cell counter racks directly, eg.: Bayer, Sysmex, Beckman Coulter, Abbott, ABX, etc… 92

Tubes lock

TUBES LOCK

Tubes unlock 93

SAMPLE ID WITH EXTERNAL BAR-CODE READER

Tube identification With the bar code reader the ID patient is automatically assigned to the ESR result Tube introduction into the rack 94

AUTOMATIC ID PATIENT WITH QUERY HOST SOFTWARE

Racks used: Internal Bar Code Reader Sample Identification with automatic selection of the tubes requiring the ESR test both with dedicated racks.

95

WORKFLOW

Rack blocking Tubes are blocked with an elliptic movement Rack loading Open the front door and insert the rack into the instrument.

Close the front door and press the START key.

96

RACK ADAPTOR FOR BECKMAN COULTER

97

RACK ADAPTOR FOR BAYER

98

RACK ADAPTOR FOR SYSMEX - ABBOTT - ABX

99

The most recent work made by an Hospital in Verona, Italy, is an important example of TLA (Total Lab Automation), published at Euromedlab, Barcelona 2003, in which the author declares what you can see in the poster (500 tubes of ESR per day not using, as in the past, a dedicated sodium citrate tube).

100

The picture of the lab shows an integration of Sysmex cell counter and 2 TEST1 connected to the LIS. For the purpose of this automation the workflow is now available to perform the ESR test using directly the cell counter racks of Abbott, ABX, Beckman Coulter, Bayer and Sysmex. Thus, TEST1 is loaded with special rack adaptors to contain the original hematology racks with an internal bar code reader selecting the ESR test connected to LIS. I remind you no other author, as far as I can know, has published anything new with the old method and with the old reader systems.

101

102

INSTRUMENTS IN THE WORLD

updated 26.04.04

102

103

T O T A L INSTRUMENTS IN THE WORLD TEST 1 734 41 40 21 20 13 12 9 5 6 1

TEST1 MicroTEST1

308 48 75 42 75 55 21 31 17 10 6 41 17 32 3 1 5 13 1 3 4 5 9 4 3 1 3 1 1 1 1 2 1 2 1 1 1 1 1 1 1 1 1

Roller 20

8 14 3 2 3 1 2 2 2 1 3

Country

Italy South Africa Spain Korea Brasil Turkey China Belgium Portugal Australia Germany Colombia Ireland/UK Israel New Zealand Slovenia Uruguay Costa Rica Malaysia Guatemala Nederlands India Hungary Syria Norway U.S.A.

Nicaragua El Salvador Panama Greece Indonesia Singapore Vietnam Chile Japan Micro TEST 1 T O T A L 285

103

Sales of TEST 1 tests 12.000.000

10.000.000

10.161.000

104

8.000.000

7.104.000

6.000.000

4.000.000

2.000.000

819.000

2.218.000

3.058.500

4.580.500

0 1998 1999 2000 2001 2002 2003

104

Download