1. Unexpected Antibody Screening & Identification 제품 소개
주식회사 유니온랩

2. DIAGNOSTIC GRIFOLS
Spain 에 본부를 둔 약 50년 역사의 수혈의학관련 제품 및 면역혈청진단 및 치료제 전문기업입니다.
1998년, 세계 최초로 전자동 혈액은행 검사 장비 WADiana 를 출시하였습니다.
혈액은행검사 자동화 분야에 있어서는 가장 앞서 있는 선두주자입니다.
DiaMed 사에는 ID Gel Station 이란 이름으로, J&J Ortho 에는 ProVue 라는 이름으로 자동화 장비를 OEM 공급하고 있습니다.
2006년 현재, 전세계에 1,700 여대의 장비가 가동 중에 있습니다.
WA Diana 는 미국 FDA 의 승인을 받은 유일한 혈액은행검사 자동화 장비입니다.

3. Gel Column Technique 안정한 반응 표준화 기법 높은 감도 보다 분명한 결과 간편한 취급
항글로불린검사시 세척과정 불필요
적은 검체량
자동화 용이

4. Grifols DG Gel ® Cards 종류
Cat #
DG Gel
Configuration 1
210310
DG Gel ABO/Rh
A B AB D CDE Ctrl A1 B 2
210311
DG Gel ABO/Rh (2D)
A B AB D D' Ctrl A1 B 3
210313
DG Gel ABO-CDE
A B AB D C E A1 B 4
210312
DG Gel Confirm
2 x (A B D Ctrl) 5
210314
DG Gel Anti-D
4 x (D' Ctrl) 6
210315
DG Gel Rh
2 x (C c E e) 7
210316
DG Gel Rh+Kell
D' C Cw c E e Kell Ctrl 8
210319
DG Gel Coombs
8 x (AHG) 9
210320
DG Gel Neutral
8 x (N) 10
210321
DG Gel Neutral/Coombs
N N N N AHG AHG AHG AHG 11
210317
DG Gel Neonatal
A B AB D D' Ctrl IgG AHG 12
210323
DG Gel DC Scan
2 X (AHG IgG CD3 Ctrl) 13
210318
DG Gel CT
A B D Ctrl N N AHG AHG 14
210324
DG Gel AB
4 X (A B) 15
210322
DG Gel IgG
8 x (IgG)

5. Unexpected Antibody Test 용 진단제품
DG Gel Coombs Card
마이크로 컬럼법을 사용하는 혈액형판정용 카드제품중 Sephadex Gel column 방식은 Micro glass bead column 방식보다 specificity 가 뛰어납니다. 또, Gel column 방식은 Glass bead column 방식에 비해 false positive 율이 거의 없습니다.
카드 한 개당 8개의 column (4 환자분)으로 구성되어 있습니다.

6. Unexpected Antibody Test 용 진단제품
SeraScan Diana - 비예기항체 선별용 Screen cells
제품 사용기간이 제조일로부터 8주로서 타제품들 (6주)보다 사용기간이 길어 손실율이 적습니다.
제품 자체가 LISS (Low Ion Strength Solution) 처리되어 있어 항체 검출율을 높이고 별도의 LISS 용액시약이 필요없으며 따라서 검사과정도 간단합니다.
검출 가능한 비예기항체의 종류는 국제표준인 23종의 항체를 포함합니다. D,C,E, c, e, K, k, Kpb, Jsb, M, N, S, s, Fya, Fyb, Jka, Jkb, Lea, Leb, Lub, P1, Xga

7. Unexpected Antibody Test 용 진단제품
SeraScan Diana - 비예기항체 선별용 Screen cells
Cw 항체를 항상 포함하고 있습니다.
가장 흔히 검출되는 Rh, Duffy, MNS, Kidd 시스템을 망라하고 있습니다.
Homozygous 인 D, C, c, E, e, Fya, Fyb, Jka, Jkb 를 각각 검출합니다.
R1R1 와 R2R2 profiles 이 있습니다.

8. Unexpected Antibody Test 용 진단제품
IdentiSera - 비예기항체 동정용 cells
비예기항체 동정을 위한 적혈구 시약으로 11 vial로 구성되어 있습니다.
정확히 동정되지 않는 특수케이스를 위해 추가동정을 할 수 있는 4 vial 이 별도로 준비되어 있습니다. (IdentiSera Extend)
세 개의 Kell-positive cells 을 함유하고 있습니다.
Homozygous 항체 - M, N, S, s, C, c, E, e, D, Lub, k, Kpb, Jsb, Fya, Fyb, Jka, Jkb 모두 동정 가능합니다.

9. Unexpected Antibody Test 용 진단제품
IdentiSera - 비예기항체 동정용 cells
Minimum two homozygous cells for: Fya, Fyb, Jka, Jkb, S, s, M, N
Presence of: P1 negative, Le(a-b-), 2 cells Le(a+)
한 판넬에 3 R1R1 cells, 4 rr homozygous cells, 그리고 보조적 Jk, Fy, MNS cells 이 모두 있습니다.
With presence of anti-D, anti-C or anti-E, a 2nd antibody of Kidd, Duffy, MNS or Kell can be detected
4 possible complementary combinations for Fy and Jk

10. Identisera + Identisera Extended
Total 15 cells – 비예기항체 동정이 완벽합니다.
Control
1 patient = 2 cards = 15 cells + 1 Control

11. DG Gel® + Identisera + Identisera Extended
최대수의 homozygous 항원과 보조적 적혈구항원의 조합으로 비예기항체의 검출과 동정을 완벽히 수행 할 수 있습니다.
Maximum efficiency and information per card and test
Optimization of the unique 8 column format
Grifols Clear Card Technology

12. Diana System Equipment
Diana Incubator
24 Gel cards 동시장착가능
(24 cards X 4 Pt’s/card = 96 Pts)
Timer 내장
Incubation 시간 : 37도 15분간
열전도에 의한 가온법
실험 튜브도 가온 가능

13. Diana System Equipment
DianaFuge
동시 24 Gel cards 원심분리 가능
1100 rpm 에서 9분간 원침

14. Unexpected Antibody Test 용 진단제품
우수한 성능
기존 제품과의 평가에서 가장 우수한 성적을 보였습니다. (2005 AABB poster 및 2006 논문 full text 참조)
(J. Cid et al.
Comparison of three microtube column agglutination systems for antibody screening: DG Gel, DiaMed-ID and Ortho BioVue
Transfusion Medicine 2006, )
간편한 2 step 법
혈청 25uL 분주
선별용 혈구 50uL 분주 (LISS 사용 step 없음)
 Incubation + Centrifuge
경제성
명확한 결과 및 정확한 검사
Gel card 한 개당 4환자 검사

15. Advantages of DG Gel Card
8-column format
분명한 결과
간편한 사용법
쉬운 판독
사전 수혈검사시
높은 신뢰성

16. Mean±SD score of antibody titration
Joan Cid, Núria Nogués, Rosa Montero, Marta Hurtado, Albert Briega and Rafael Parra.
Blood and Tissue Bank. Barcelona, Spain.
Comparison of three microtube column agglutination systems for antibody screening: DG Gel, DiaMed-ID and Ortho BioVue
INTRODUCTION
Antibody screening is performed to detect clinically relevant red blood cell (RBC) antibodies in transfusion recipients and blood donors. The classical low-ionic-strength-solution (LISS) tube indirect antiglobulin test (IAT) for detecting clinically significant antibodies has been replaced in many countries by microtube column agglutination systems, described for first time in 1988 and published in 1990 (1). The first microtube system was manufactured and distributed by DiaMed-ID (Switzerland), followed by Otho BioVue (Otho Diagnostic GmBH, Germany). Recently, a new microtube column agglutination system has been launched on the market by Diagnostic Grifols, S.A. (Barcelona, Spain) with the name of DG Gel.
The aim of the present study was to evaluate the diagnostic accuracy of this new system, to analyze the antibody reactivity and to compare the data with the two cited well-established methods.
METHODS
Samples. We collected 3,024 specimens: 2,408 (79.6%) were plasma samples obtained from EDTA tubes and 616 (20.4%) were serum samples. We obtained 2,422 (80.1%) samples from blood donors in our blood bank; 382 (12.6%) samples from transfusion recipients and 220 (7.3%) from pregnant women in Hospital Vall d’Hebron (Barcelona, Spain). We stored all these samples in the refrigerator at 2-8 ºC and we processed them before 72 hours from extraction. We additionally obtained 100 samples containing antibodies of known specificity from the immunohematology laboratory of our blood bank. These samples had been stored in a frozen state at -30 ºC and were thawed immediately before processing.
Test RBCs. The RBCs used for the antibody screening and for the panel identification were obtained from our selected blood donors. These RBCs were suspended to final concentration 0.8 % in the corresponding modified LISS provided by the manufacturers according to their instructions as follows. We calculated the conductivity of these solutions.
DG Gel: 50 µL of RBCs in diluent provided by Diagnostic Grifols (Cond.: 5.09 mS/cm) + 25 µL of sample. Incubation: 15 min, 37 ºC. Centrifugation: 9 min.
DiaMed-ID: 50 µL of RBCs in ID-CellStab (Cond.: 5.89 mS/cm) + 25 µL of sample. Incubation: 15 min, 37 ºC. Centrifugation: 10 min.
Ortho BioVue: 50 µL of RBCs in Ortho BLISS (Cond.: 3.03 mS/cm) + 40 µL of sample. Incubation: 10 min, 37 ºC. Centrifugation: 5 min.
RESULTS
Antibody screening
We performed the antibody screening of 3,024 fresh samples with the three microtube systems and we obtained concordant results in 3,019 (99.83%) and discrepant results in 5 (0.17%). Table 1 shows the specificities of the antibody after the identification steps using 11-cell panel and additional techniques, e.g. papain-treated reagent RBCs and IAT with polyethylenglycol (PEG) tube technique (2).
Antibody titration
We titrated 100 frozen samples containing antibodies with known specificities. The score mean and standard desviation of the antibody titration for each microtube system were the 34.31±26.7 for DG Gel, 30.3±27.76 for Diamed-ID, and 37.38±28.68 for Ortho BioVue (see Table 2).
The antibody titration score was higher with the DG Gel when compared with DiaMed-ID in samples containing anti-K (46.23 vs ; p<0.05) and in samples containing anti-M (11.88 vs. 5.38; p<0.05). No significant differences were seen in other antibodies specificities.
CONCLUSION
All three microtube column agglutination systems work well showing a high estimated sensitivity and specificity.
REFERENCES
1. Lapierre, Y. Rigal, D., Adam, J. Josef, D., Meyer, F., Greber, S. and Drot, C. (1990) Transfusion, 30,
2. AABB (2002) Technical Manual, 14 th edition, Beshesda.
Table 1. Antibody detected by the three systems using 2 screening cells.
Mean±SD score of antibody titration
37.38
30.3
34.31
100
Total 74 86 1
Anti-Xga 43 33 46
Anti-Vel 24 17
Anti-Ge:2
4829.69
41.5037.47
48.5034.64 2
Anti-PP1PK
20.6714.18
12.3310.69
13.3313.79 3
Anti-H/HI/I
161.41
5.503.53
11.502.12
Anti-Leb
87.07
41.41
80
Anti-Lea
4121.15
30.4020.73
3417.01 5
Anti-S
29.6718.14
24.6727.79
2417.08
Anti-s 25 39
Anti-N
16.887.69
5.382.82
11.884.73 8
Anti-M 19 12
Anti-Kpb
2614.14
16.5012.02
17.506.36
Anti-Kpa
50.3137.60
44.3836.72
46.2336.73 13
Anti-K
5217.37
45.4511.14
50.2514.68 4
Anti-Fyb
35.8026.01
24.4024.70
3424.87
Anti-Fya
210
194.24
Anti-Jkb
22.4316.20
15.8611.65
20.5714.50 7
Anti-Jka
3618.38
248.48
38.5019.09
Anti-Ch
167.21
14.679.50
15.339.6
Anti-Cw
130
200
Anti-e
34.3029.33
33.3028.98
37.4023.14 10
Anti-E
28.2224.80
2319.55
23.6719.44 9
Anti-c
70±19.71
60.75±15.54
63±16.45
Anti-C
82.29±29.35
74.43±23.86
74.14±24.23
Anti-D
Ortho BioVue
DiaMed-ID
DG Gel n
Specificity
False negative
Kell
Duffy
Kidd
3024
Antibody specificity
2998
3003
3000
Negative
False positive
Panaglutinin
Nonspecific
Anti-Jkb/-K
Anti-E/c/panaglutinin
Anti-C/-D
Mixed
Lewis
MNS 6 Rh
Number of Samples Detected
Blood Group
System
Table 2. Samples containing antibodies of known specificities used for antibody titration.
99.94%
( )
99.9%
( )
100%
( )
Efficiency
( )
PNV(b)
98.41%
( )
(97-100)
( )
PPV(a)
99.93%
( )
97.58%
( )
Sensitivity
Table 3. Estimated diagnostic accuracy. In parenthesis the 95 % CI. (a) PPV: predictive positive value, (b) PNV: predictive negative value.
Sensitivity and Specificity
In order to obtain a comparative value between the new test and the other two we calculated the estimated diagnostic accuracy for each system (see Table 3). For calculating estimates of these statistical measures we include the 100 positive samples and we reported the confidence intervals (CI) for a sample size of 3,124.