到馬偕報告

1. Are manual Polybrene ( MP) only antibodies clinical
significance ? Serological View.
張志昇
馬偕紀念醫院
2014/11/5

2. Outline
 Definition of clinical significant antibody & MP only allo-antibody
 Background
 Literature review
 Purpose
 Case reports
 Laboratory finding
 Conclusion
小張的報告 Page 2

3. Definition of clinical significant
 An antibody is considered clinically significant if it is capable of inducing either hemolytic disease of the
fetus/newborn (HDFN) or accelerated red cell destruction.
 A term used to describe the likelihood a particular blood group antibody that can cause one of two
problems: 1) Hemolytic transfusion reactions (accelerated red blood cell destruction), or 2) Hemolytic
disease of the fetus/newborn (HDFN).
 In general, red cell antibodies that react at body temperature (37 C) are more likely to be significant
 Most warm antibodies are IgG class rather than IgM.
 Antibodies that can cross the placenta and circulate in the fetus are obviously more likely to cause
HDFN, so IgG antibodies are more likely than IgM to be significant from that perspective. However, this
does NOT mean that all IgM antibodies are insignificant.
 One of the clues as to whether or not an antibody will be "significant" is whether or not the antibody
reacts at body temperature (37C). The optimal reaction temperature of an antibody is not always
predictive of its clinical significance, but it can definitely guide you in the right direction
小張的報告 Page 3

4. Definition of Manual Polybrene (MP) only allo-antibodies
 Manual polybrene (MP) : Modified manual Polybrene method : Marie Lin林媽利醫師所引進及改進
適用於台灣地區的方法，和文獻上使用的Manual Polybrene test ( MPT) 或Low ionic Polybrene
( LIP )略有不同。
 台灣地區常最使用的主要抗體篩檢方法為manual Polybrene法，臨床上常見紅血球異體抗體在傳統
classical AHG法無法偵測，但使用Manual polybrene法卻可以偵測到，此類紅血球抗體定義為MP
only allo-antibodies。
 Manual polybrene法反應溫度在室溫，又台灣使用的modified manual Polybrene並沒有使用anti-human
IgG antibody，通常對冷型抗體反應較傳統classic AHG 反應較敏感（sensitivity較高）．
小張的報告 Page 4

5. Outline
 Definition of clinical significant antibody & MP only allo-antibody
 Background
 Literature review
 Purpose
 Case reports
 Laboratory finding
 Conclusion
小張的報告 Page 5

6. Background
 導入血庫自動化檢驗儀器前，曾評估MP法和CAT法的差異性結果如下:
使用傳統MP法執行抗體篩檢之異體不規則抗體檢出率約為0.86％(506 / 59095)(抗體/總檢體數)，其
中抗體鑑定結果為MP only 約占18％(94/506)；而CAT法之異體不規則抗體檢出率約為1％(520 /
51727)，抗體鑑定結果為MP only約為3.5％(18/520)。分析抗體特異性，MP法異體抗體之較高比例
為anti-Mia(42％), anti-E(&c)(31％), anti-Lea(6％) , 而CAT 法之較高抗體分析為anti-Mia(43％), anti-
E(&c)(25％) , anti-M(8％)
 使用血庫自動化檢驗儀器後，經常發現使用CAT法抗體篩檢陰性，但使用ＭＰ交叉試驗法卻為交叉不
合，抗體鑑定結果為anti-e, anti-Jka, anti-E, anti-Mia等。且多例為輸血後檢體才檢驗出，疑為輸血產
生異體抗體。
 常見的MP only allo-antibodies經常為冷反應型(cold reactive allo-antibodies), 如anti-Mia, anti-E,
antiP1, anti-M, anti-Lea, anti-Leb, anti-I等，但也偶見如anti-Jka, anti-Jkb, anti-e, anti-c等抗體。本研
究希利用血清學方法分析探討MP only allo-antibodies的血清特性。
小張的報告 Page 6

7. 臨床抗體篩檢CAT法與MP法檢出率及檢出抗體
特異性之比對分析
簡妙如林美華林霈文廖文綱詹如萍王綉棉劉斐雲羅仕錡
國立台灣大學附設醫院檢驗醫學部
Background Results
Table 1.MP & CAT檢出抗體陽性結果比較表
Table 2.MP & CAT檢出抗體特異性結果分析表
Anti-Lea
Conclusions
Reference
Column Agglutination Technology (CAT)是1990年初發展出的新技術用
以檢測抗原抗體間的反應，本院於2001年開始將CAT法應用在抗體
鑑定，直至2012年9月中才全面以自動化血型分析儀應用於抗體篩
檢，取代Manual Polybrene (MP)法。根據CAP 2013 Transfusion
medicine JB survey ，結果分析顯示Gel testing佔所有Antibody
screening 方法約66%(2290/3492) ， Tube method 約31%(1075/3492);
因此以CAT執行抗體篩檢已漸漸成為國際趨勢。
使用傳統MP法執行抗體篩檢之異體不規則抗體檢出率約為0.86％
(506 / 59095)(抗體/總檢體數)，其中抗體鑑定結果為MP only 約
占18％(94/506)；而CAT法之異體不規則抗體檢出率約為1％(520 /
51727)，抗體鑑定結果為MP only約為3.5％(18/520)。分析抗體特
異性，MP法異體抗體之較高比例為anti-Mia(42％), anti-E(&c)(31
％), anti-Lea(6％) , 而CAT 法之較高抗體分析為anti-Mia(43
％), anti-E(&c)(25％) , anti-M(8％)。
1.檢出率：檢體樣本數以回溯方式分析比較自2011年10月至2012年
9月所有輸血及備血檢
體以MP法執行抗體篩檢
及2012年10月至2013年
8月中以CAT法執行抗體
篩檢之總檢體量(為分
母)，及新檢出之異體抗
體數(為分子)，分析比
較抗體檢出率。
2.抗體特異性：新檢出抗體以CAT為主要抗體鑑定法，其他則為MP、
3 phases AHG method；
本次分析抗體以新檢出
異體抗體為主，已鑑定
過之抗體及自體抗體則
不重覆計算，多重抗體
則可重覆計數。
AHG抗體
MP Method CAT
Anti-M
分析結果顯示：CAT法之異體不規則抗體檢出率較MP法高；Rh類
抗體有較明顯比例差異，探討原因可能為MP法對Rh類抗體較
sensitive，所以檢出較高MP only之Rh類抗體約20％( 41/198)。但就
anti-M及anti-Lea之檢出結果則為CAT法檢出較多，其原因可能為
部份較弱之anti-M及anti-Lea抗體反應特性在MP法判讀時較容易散
開，不易被檢出。其他具臨床意義之抗體如anti-Jka, -Jkb, -S及-Fyb
則在CAT法(6.3％)較MP法(4.7％)有較佳之檢測結果。
結論：抗體檢出率CAT法檢出率為(1％)較傳統MP法(0.86％)好，而
就檢出抗體分析，以CAT法screening出較多AHG反應之抗體如anti-
Jka, -Jkb, -S及-Fyb。自動化血型分析儀之CAT法作業模式可以將實
驗室間人員操作及判讀上差異降低，提高抗體之檢出。此外Anti-
Mia, anti- E(含c)為本院最常見之異體不規則抗體，此與國內之其他
發表文獻結果ㄧ致。
Objectives
MP是臺灣最常用的抗體篩檢法，其特點是反應時間短，對冷型抗
體及Rh類抗體之敏感度高，但對於一些反應溫態為AHG的抗體如
anti-Kidd, anti-Duffy & anti-Kell則較不敏感； CAT法則對AHG抗體有
較高的敏感性，以自動化血型分析儀可將操作及判讀更標準化，可
減少人為操作及判讀上的誤差。本研究目的為比對CAT與MP此兩種
不同抗體篩檢方法，並分析其免液凝集敏感度對抗體篩檢檢出率及
檢出抗體特異性。
Figure 1.MP抗體陽性處理流程
Abbrev.
Ab: antibody; Ab SRC: antibody screen; AC: autocontrol; Ag: antigen; DAT:
Direct Coombs’ test; IS: immediate spin; XM: crossmatch
MP 法抗體篩檢CAT 法抗體篩檢
檢體收集時間2011.10-2012.09 2012.10-2013.08
月數12個月10.5個月
備輸血檢體數59095 51727
抗體篩檢陽性個數506 520
陽性率0.86% 1%
MP only抗體陽性率94/506=18% 18/520=3.5%
RH類及其他類抗體
MP Method CAT Method
Specificit
y
Antibody frequency
Mia 236 (46.6%) 246 (47.3%)
E&c 174 (34.4%) 142 (27.3%)
C&e 24 (5%) 13 (2.5%)
D 3 (0.6%) 3 (0.58%)
Dia 6 (1.2%) 13 (2.5%)
Wra 4 (0.8%) 2 (0.38%)
% 447/506
(88.3%)
419/520
(80.6%)
冷型抗體
MP Method CAT Method
Specificity Antibody frequency
M 30 (5.9%) 47 (9.4%)
Lea 34 (6.7%) 42 (8.1%)
Leb 7 (1.4%) 18 (3.5%)
P1 14 (2.8%) 9 (1.7%)
% 85/506 (16.8%) 116/520 (22.3%)
Method
Specificit
y
Antibody frequency
Jka 9 (1.8%) 14 (2.7%)
Jkb
4 (0.8%)
5 (0.96%)
Jk3
1 (0.2%)
0
Fyb 3 (0.6%) 9 (1.7%)
S 8 (1.6%) 5 (0.96%)
% 25/506
(4.9%)
33/520
(6.3%)
Figure 2. MP法檢出抗體
Anti-Mia
Anti-E & c
Figure 3. CAT法檢出抗體
Anti-Mia
Anti-E & c
Manual polybrene Ab SRC
(AC not performed)
CAT panel & AC
CAT panel &
AC negative
Repeat MP Ab
SRC
No
further
MP Ab
panel
IS XM
Determine Ab
Completely XM
Ag negative units
CAT panel positive
& AC positive
Follow work-up for
CAT panel positive &
AC positive
CAT panel poisitve
& AC negative
CAT completely
XM with Ag
negative units
No further
work-up
IS XM
Negative positive
Transfused in
three months ,
then perform
elution test.
Determine Ab
and phenotype
paitent
Byrne T, Nolan T, O'Donnell R, Br J Biomed Sci. 1996 Sep;53(3):193-5.
V. Weisbach, A. Ziener, R. Zimmermann, TRANSFUSION 1999;39:1045-
1050.
Methods

8. 以自動血型檢測機AutoVue與試管法偵測血型及Manual
Polybrene法偵測不規則抗體之平行測試
林美華戴宜娥簡妙如廖美玟劉斐雲羅仕錡
國立台灣大學附設醫院檢驗醫學部
簡介
及交叉試驗，其目的是提供安全的血品，防止受血者產生溶血性
輸血反應，以達輸血治療之目的！其中不規則抗體篩檢的目是在
最短時效內，儘可能篩檢出越多具有臨床意義的抗體，並減少篩
檢出不具臨床意義抗體。因此選擇一適當方法做輸血前檢驗更顯
其重要性。
agglutination technology)和傳統試管法(tube method)進行血型
和不規則抗體篩檢之平行測試。AutoVue CAT之不規則抗體篩檢原
理為改良傳統三相法，使步驟簡化，以達到AABB Standard
5.13.3抗體篩檢須包括37℃之AHG test的要求。
試劑與方法
Blood Type
1. Tube Method:
• Anti-A & Anti-B: NOVACLONE, Murine Monoclonal
• Anti-D: NOVACLONE, Anti-D IgM+IgG
Monoclonal Blend
• A cells & B cells: 台塑抗體篩檢細胞組
2. Column Agglutination Technology(CAT):
• Ortho BioVue System Blood Grouping
Reagents(ABO-Rh/Reverse Grouping Cassette)
• A cells & B cells: 台塑抗體篩檢細胞組
Antibody Screen
1. Manual Polybrene(MP):
ABScr 3 Poly
結果
1.血型檢驗在手工試管法與AutoVue之間的比對
取98隻EDTA管抽取之全血，用手工試管法與兩台AutoVue機台
做血型檢驗的比對。
92隻臨床隨機Routine檢體結果
100% 達到一致，為了檢測在
弱反應血型上之差異，選取
了6個在tube method弱反應
血型，有4隻AutoVue與tube
method結果不一致。
不一致結果4個當中檢體A1的Rh typing於兩台AutoVue機台結果顯
示陰性結果，但手工試管法呈現1+的弱陽性反應；
檢體A2於兩台AutoVue機台ABO forwardtyping反應太
弱顯示？結果，檢體A4的ABO forwardtyping在兩台
AutoVue機台顯示mixfield與？的結果，reverse
typing於其中一台AutoVue有fibrinogen結果干擾判
讀，與B cells反應也為4+，此檢體Rh typing於兩台
AutoVue機台無法判讀結果，但是以試管法可以清楚
判讀出血型。檢體A6於兩台AutoVue機台reverse
typing皆在A1 cells有很強的反應。
AutoVue (5065) AutoVue (5066) tube method
sample Anti-A Anti-B Anit-D Control A1 cells B cells Anti-A Anti-B Anit-D Control A1 cells B cells Anti-A Anti-B Anit-D A1 cells B cells
A1 0 0 0 0 3 4 0 0 0 0 3 4 0 0 1 3 3
A2 0 ? 4 0 0 0 0 ? 4 0 0 0 0 1 4 0 0
A3 0 0 4 0 0 2 0 0 4 0 0 2 0 0 4 0 3
A4 4 MF ? 0 FIB 4 4 MF ? 0 0 0 4 4 0 0 0
A5 0 0 4 0 3 3 0 0 4 0 3 3 0 0 4 1 +/-
A6 0 0 4 0 3 4 0 0 4 0 3 4 0 0 4 0 4
2.抗體篩檢在MP法與AutoVue之間的比對
抗體篩檢結果比較
AutoVue (5065)IgG
card做65個檢體比對。36個
檢體在MP法表現與AutoVue
AutoVue (5066)IgG
兩種卡片篩檢皆為Negative
(僅sample 2 之MP法表現為
AutoVue (7272)Poly
Negative，在IgG card為
部分)。而MP法Positive的27個
檢體，有13個檢體在CAT法表現Negative或是不同的反應結果(黃
不規則抗體篩檢的比對
則採用MP(Manual Polybrene)
法與AutoVue CAT之poly(poly-specific
AHG)card ＆ IgG
色部分)，進一步抗體鑑定後發現其差異在於，CAT
對於較不具臨床意義之IgM冷型抗體反應較弱或不反
應；但對較具臨床意義之IgG抗體CAT法反應價數通
常比MP強。本院輸血前檢驗檢體為EDTA管採集的全
血，因此在AutoVue poly card ＆ IgG card反應強
度結果無明顯差異。
輸血前檢驗，包括ABO/RH血型、不規則抗體篩檢、抗體鑑定
本院以AutoVue自動化血型鑑定之柱狀凝集法CAT(column
0.8% cells & Serum
Air Gap
Antihuman Globulin
and Diluent
Glass Beads
AutoVue-5065 (AHG IgG) Autovue-5066 (AHG IgG) AutoVue-原機(AHG POLY) MP法
MP method
POS NEG 未測
POS 20 1 2
NEG 7 35 0
未測0 0 0
POS 14 1 2
NEG 1 35 0
未測12 0 0
POS 18 0 2
NEG 5 34 0
未測4 2 0
No. Surg1 Surg2 Surg3 interpretation Surg1 Surg2 Surg3 interpretation Surg1 Surg2 Surg3 interpretation Surg1 Surg2 Surg3 interpretation Remark
1 0 3 0 ABscr POS 0 3 0 ABscr POS 0 3 0 ABscr POS 0 2 0 ABscr POS anti-E+Mia
2 0 3 0 ABscr POS 0 3 0 ABscr POS 0 0 0 ABscr NEG 0 0 0 ABscr NEG anti-Mia
3 0 0.5 0 ABscr POS 0 2 0 ABscr POS 0 2 0 ABscr POS 0 2 0 ABscr POS anti-Mia
4 0 2 0 ABscr POS 0 2 0 ABscr POS 0 2 0 ABscr POS 1 1 1 ABscr POS anti-Mia
5 2 2 0 ABscr POS 2 2 0 ABscr POS 2 2 0 ABscr POS 1 1 1 ABscr POS anti-E+Mia+Cold
6 0 2 0 ABscr POS 0 2 0 ABscr POS 0 3 0 ABscr POS 0 2 0 ABscr POS anti-Mia
7 0 4 0 ABscr POS 0 4 0 ABscr POS 0 4 0 ABscr POS 0 3 0 ABscr POS anti-Mia
8 3 1 0.5 ABscr POS 3 0.5 0.5 ABscr POS 3 1 0.5 ABscr POS 0 1 1 ABscr POS anti-E+c+Jkb
9 4 0 0 ABscr POS 4 0 0 ABscr POS 4 0 0 ABscr POS 2 0 0 ABscr POS anti-E
10 0.5 0.5 0.5 ABscr POS 0.5 0.5 0.5 ABscr POS 0.5 0.5 0.5 ABscr POS 1 1 1 ABscr POS warm auto
11 0 0 0 ABscr NEG 0 0 0.5 ABscr POS 0 0 0 ABscr NEG 0 0 1 ABscr POS cold auto
12 0 0 0 ABscr NEG x 0 0 0 ABscr NEG 2 0 0 ABscr POS anti-c+E
13 0 0 0 ABscr NEG 0 0 0 ABscr NEG 0 0 0 ABscr NEG 0 1 0 ABscr POS anti-Mia
14 0 0 0 ABscr NEG 0 0 0 ABscr NEG 0 2 0 ABscr POS Anti-Mia
15 0 2 0 ABscr POS 0 3 0 ABscr POS 0 2 0 ABscr POS Anti-Mia
16 0 2 1 ABscr POS 0.5 2 2 ABscr POS 0 3 3 ABscr POS Anti-C+e+Mia
17 0 0 0 ABscr NEG 0 0 0 ABscr NEG 1 1 1 ABscr POS Anti-Mia+Wra+Dia+M+War,
18 0 0 0 ABscr NEG 0.5 0.5 0 ABscr POS 1 0 0 ABscr POS Anti-E+c
19 0 0 1 ABscr POS 0 (WLL) 0 1 ABscr POS 0 0 1 ABscr POS Lea
20 0 0 0 ABscr NEG 0.5 ? 0 ABscr POS 2 0 0 ABscr POS Anti-E+c
21 0 3 0 ABscr POS 0 3 0 ABscr POS 0 3 0 ABscr POS Anti-Mia
22 0 2 0 ABscr POS 0 1 0 ABscr POS 0 1 0 ABscr POS Anti-Mia
23 0.5 3 0 ABscr POS 0 3 0 ABscr POS 1 2 0 ABscr POS Anti-E+c+Mia
24 3 3 3 ABscr POS 3 3 3 ABscr POS 1 1 1 ABscr POS Anti-Mia+M
25 2 1 1 ABscr POS 2 2 1 ABscr POS x 2 2 2 ABscr POS anti-c+Jkb+warm auto
26 2 0 0 ABscr POS 2 0 0 ABscr POS x 2 2 2 ABscr POS anti-c+E / cold auto anti-I
27 0 0.5 0 ABscr POS 0 0.5 0 ABscr POS x 0 1 0 ABscr POS anti-Lea
28 0 3 4 ABscr POS 0 3 4 ABscr POS x 0 1 0 ABscr POS anti-Mia
結論
此次評估結果發現，AutoVue CAT和傳統試管法進行血型比對結果幾乎達到一致性；但在不規則抗體篩檢之比對差異較大，主要是因MP
法會篩出的不具臨床意義之IgM冷型抗體！輸血前檢驗自動化除了可標準化減少人為操作及判讀上之誤差，最重要是可以減少文書抄寫錯誤，
所以已漸漸成為國際驅勢！

9. 小張的報告 Page 9

10. 小張的報告 Page 10
MP 法抗體篩檢CAT 法抗體篩檢
檢體收集時間2011.10-2012.09 2012.10-2013.08
月數12個月10.5個月
備輸血檢體數59095 51727
抗體篩檢陽性個數506 520
陽性率0.86% 1%
MP only抗體陽性率94/506=18% 18/520=3.5%

11. 臨床抗體篩檢CAT法與MP法檢出率及檢出抗體特異性之比對分析
小張的報告 Page 11
RH類及其他類抗體
MP Method CAT Method
Specificity Antibody frequency
Mia 236 (46.6%) 246 (47.3%)
E&c 174 (34.4%) 142 (27.3%)
C&e 24 (5%) 13 (2.5%)
D 3 (0.6%) 3 (0.58%)
Dia 6 (1.2%) 13 (2.5%)
Wra 4 (0.8%) 2 (0.38%)
% 447/506 (88.3%) 419/520 (80.6%)
AHG抗體
MP Method CAT Method
Specificity Antibody frequency
Jka 9 (1.8%) 14 (2.7%)
Jkb
4 (0.8%)
5 (0.96%)
Jk3
1 (0.2%)
0
Fyb 3 (0.6%) 9 (1.7%)
S 8 (1.6%) 5 (0.96%)
% 25/506 (4.9%) 33/520 (6.3%)
冷型抗體
MP Method CAT Method
Specificity Antibody frequency
M 30 (5.9%) 47 (9.4%)
Lea 34 (6.7%) 42 (8.1%)
Leb 7 (1.4%) 18 (3.5%)
P1 14 (2.8%) 9 (1.7%)
% 85/506 (16.8%) 116/520 (22.3%)

12. Outline
 Definition of clinical significant antibody & MP only allo-antibody
 Background
 Literature review
 Purpose
 Case reports
 Laboratory finding
 Conclusion
小張的報告 Page 12

13. Literature review-1
 Manual hexadimethrine bromide (Polybrene) tests (Polybrene in low-ionic medium) were used in
parallel with manual low-ionic-strength solution (LISS) procedures for the routine testing of patient
samples referred to a general hospital blood bank. Of 5646 consecutive sera tested, 5167 (91.5%)
did not react with either technique; 320 sera (5.7%) reacted in both methods. The Polybrene
technique detected 63 antibodies which did not react in the LISS methods. One hundred sera did
not react in the Polybrene test, but did react in the LISS methods. Sera showing discrepant results
between the 2 methods were further tested in a reference laboratory. Polybrene tests appeared to
be better in avoiding reactions due to clinically nonsignificant antibodies. The LISS methods
appeared to be more sensitive in detecting antibodies of potential clinical significance.
Transfusion. 1985 Mar-Apr;25(2):145-8.
小張的報告 Page 13

14. Literature review-2
 To explore the comparison of values of microcolunm gel test and polybrene test in irregular
antibody detection.Methods From 2010 to 2011,
6 720 cases with blood transfusion were test by microcolumn gel and polybrene for detecting
irregular antibody,and then positive results were tested about antibody′s specificity by anti-human
globulin test.
28 irregular antibody positive cases were detected by microcolumn gel test and positive rate was
0.42%.Only 26 cases were detected by polybrene test, and positive rate was 0.39%,there was no
statistically significant difference between two methods
《Laboratory Medicine and Clinic》2013-01
小張的報告 Page 14

15. Literature review-3
 Anti-Jka, -C, and -E in a single patient, initially demonstrable only by the manual hexadimethrine
bromide (Polybrene) test, with incompatibilities confirmed by 51Cr-labeled red cell studies.
 Analysis Antibody Identification Data from Blood Donation and Transfusion Services in Kaohsiung
2007 19:3 J Biomed Lab Sci p99~105（陳嘉文主任）
 Comparison Study of the Manual Polybrene, Gel Test, Autovue Innova and Conventional Tube Test
for Antibody Screening. 2006
 The comparative analysis and research between microcolumn gel test and polybrene test in
irregular antibody detection. Loboratory medicine 2010, Vol. 25 Issue (03): 179-181
 Evaluation of the test using polybrene for the detection of anti-erythrocyte antibodies.
 The manual polybrene test: a simple and rapid procedure for detection of red cell antibodies.
Transfusion 20:2 pg 206-11
小張的報告 Page 15

16. Evaluation of Ortho BioVue system, the DiaMed Gel system, and Gamma-ReACT
test and their comparison with Manual Polybrene test
 12 sample with positive antibody screening results by MP were
collected ans tested again with GEL, CAT, and ReACT antibody
specificities were determined.
MP GEL CAT RACT
Anti-Mia 4 2 3 2
Anti-E 2 1 2 -
Anti-M 2 1 2 -
Anti-C+e 1 1 - 1
Anti-I 3 - 2 -
Nonspecific reaction - 1 3 1
No antibody - 6 - 8

17. Evaluation of Ortho BioVue system, the DiaMed Gel system, and Gamma-
ReACT test and their comparison with Manual Polybrene test (II)
Five hundred and twenty-eight sera which were negative for red cell
antibodies by MP tests were tested with LISS, GEL, CAT,and ReACT.
One hundred and five samples with positive antibody screening results
by MP were collected and again screened with LISS, GEL, CAT, and
ReACT. Antibody was the identified using the corresponding methods.

18. Evaluation of Ortho BioVue system, the DiaMed Gel system, and Gamma-
ReACT test and their comparison with Manual Polybrene test (II) result:
MP LISS GEL CAT ReACT total
Anti-Mia 0 1 1 3 1 3
Anti-Lea 0 0 0 4 0 4
Nonspecific
antibodies
0 0 2 19 0 0
No
antibody
528 527 525 502 527 521

19. Evaluation of Ortho BioVue system, the DiaMed Gel system, and Gamma-
ReACT test and their comparison with Manual Polybrene test (II) Result-2
MP LISS GEL CAT ReACT Total
Anti-Mia 46 28 33 35 25 46
Anti-Mia 11* 2 2 11 2 14
Anti-E 36 14 25 25 18 36
Anti-C 7 1 4 4 2 7
Anti-c 7 0 2 2 0 7
Anti-e 2 0 2 2 2 2
Anti-M 10 0 0 3 0 10
Anti-Lea 7 0 3 3 1 7
Anti-D 2 0 1 1 1 2
Anti-Jka 2 0 1 1 0 2
Anti-P1 2 0 0 0 1 2
Nonspecific
0 6 3 10 3
antibodies

20. 以LISS、ONE-Step MP、ReACT、Capture方法對抗體篩檢試驗之評估
 第一組檢體為商業化抗血清，種類有Rh, Kidd, Duffy,
MNS, P, Lewis, Mia共１８個已知抗血清，做力價測試。
 第二組檢體為高雄榮總血庫收集並已做鑑定４６個病人
血清及高雄捐血中心提供２０個已作鑑定捐血者血清檢
體。
 結果：第一組Rh以One-step MP及capture方法力價倍數
高。MN, P system以one-step MP較佳，而S, Kidd,
Duffy, Kell, Lewis以capture較佳，anti-Mia則ReACT有
較高的力價。
第二組，one-step MP方法表現出最多個positive
reaction(56)。

21. 血庫作業的本土化－台灣經驗
異體抗體數量LIAT MP
Anti-E 55 42 55
Anti-E+c 10 9 10
Anti-D 5 4 5
Anti-Jka 2 2 2
Anti-Jkb 2 2 2
Anti-Jsa 2 2 2
Anti-c 4 2 4
Anti-D+C 4 3 4
Anti-e 4 2 4
Anti-C 1 0 1
Anti-C+e 1 0 1
Anti-Jk3 1 1 1
Anti-Lea 4 2 4

22. MP sensitive
2014/11/4
22
Rh MNSs Lewis Lutheran Kell Duffy Kidd
D C E c e f V VS Cw M N S s P1 Lea Leb Lua Lub K k Kpa Jsa Fya Fyb Jka Jkb Xga IS RT Eny AHG MP
1 0 + 0 + + + 0 0 0 + + + 0 0 + 0 + + 0 + 0 0 + + + + + 0 0 3+ 0 1+
2 + + 0 0 + 0 0 0 + 0 + 0 + + + 0 0 + 0 + 0 0 + 0 + + + 0 0 3+ 0 1+
3 + + 0 0 + 0 0 0 0 + + 0 + + 0 + 0 + + + 0 0 0 + 0 + + 0 0 0 0 0
4 + 0 + + O 0 0 / 0 + 0 0 + + 0 0 0 + 0 + 0 0 0 0 + 0 + 0 0 4+ 1+ 2+
5 0 0 + + + + 0 / 0 + 0 + + + 0 + 0 + 0 + 0 0 + + + + + 0 0 4+ 1+ 2+
6 0 0 0 + + + + + 0 0 + 0 + + 0 + 0 + 0 + 0 0 + 0 + 0 0 0 0 3+ 0 1+
7 0 0 0 + + + 0 0 0 + 0 0 + W 0 + 0 + + 0 0 0 0 + + + 0 0 0 3+ 0 1+
8 0 0 0 + + + 0 0 0 + + 0 + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 0 3+ 0 1+
9 0 0 0 + + + 0 / 0 + + + + 0 0 + 0 + 0 W + 0 + 0 + 0 + 0 0 3+ 0 1+
10 0 0 0 + + + 0 0 0 0 + 0 + + 0 + 0 + 0 + 0 0 + + 0 + + 0 0 3+ 0 1+
11 + + 0 0 + / 0 / 0 + 0 + 0 + 0 0 + + 0 + 0 0 + + 0 + + 0 0 0 0 0
Auto
Control
0 0 0 0 0

23. Outline
 Definition of clinical significant antibody & MP only allo-antibody
 Background
 Literature review
 Purpose
 Case reports
 Laboratory finding
 Conclusion
小張的報告 Page 23

24. Purpose
 台灣地區常最使用的主要抗體篩檢方法為manual Polybrene法，臨床上常見紅血球異體抗體在傳統
classical AHG法無法偵測，但使用Manual polybrene法卻可以偵測到，此類紅血球抗體定義為MP
only allo-antibodies。
 常見的MP only allo-antibodies經常為冷反應型(cold reactive allo-antibodies), 如anti-Mia, anti-E, anti-
P1, anti-M, anti-Lea, anti-Leb, anti-I等，但也偶見如anti-Jka, anti-Jkb, anti-e, anti-c等抗體。本研究希
利用血清學方法探討MP only allo-antibodies的血清特性。
 自從採用血庫自動化設備，偶見MP only allo-antibodies影響血庫作業程序，此探討對MP only allo-antibodies
的血清特性了解，可以增進對此類抗體的特性了解，改善輸血作業流程，並增加輸血安全。
小張的報告 Page 24

25. Outline
 Definition of clinical significant antibody & MP only allo-antibody
 Background
 Literature review
 Purpose
 Case reports
 Laboratory finding
 Conclusion
小張的報告 Page 25

26. Case report
 某月2013 : 男46Yrs HCC 因anemia Hb 6.5g/dL輸血每週2U  3，第三次輸血前AutoVue Innova
BioVue antibody screening negative, 但交叉試驗使用manual Polybrene法呈現incompatible. 經抗體
鑑定為MP only anti-Jka .
 March 某年: 女52 yrs Myoma Hb 7.2g/dL 輸血，分別於2/28, 3/26, 4/2欲輸血packed RBC各2U, 期
間似乎輸注效果不甚理想，4/2的MP 交叉試驗呈現incompatible，經抗體鑑定為MP only anti-E.
小張的報告 Page 26

27. Outline
 Definition of clinical significant antibody & MP only allo-antibody
 Background
 Literature review
 Purpose
 Case reports
 Laboratory finding
 Conclusion
小張的報告 Page 27

28. 小張的報告 Page 28

29. Result of manual Polybrene and Rh phenotype
小張的報告 Page 29

30. Manual Polybrene Test ( MPT-AGT )
小張的報告 Page 30

31. 0.02M DTT (DL-DITHIOTHREITOL) treated
小張的報告 Page 31

32. Enzyme treated panel cells reactive @ RT with poly AHG card
小張的報告 Page 32

33. Enzyme treated panel cell @ 37°C with mono-IgG card
小張的報告 Page 33

34. Direct Coombs’ test ( microscope view)
小張的報告 Page 34

35. Eluent from DAT positive cells
 仍具anti-E, anti-Jka 特性, 唯anti-E 仍呈現MP only 特性。
小張的報告 Page 35

36. 案例分享
 一泌尿科F82歲婦女血色素6.8備輸血LDPR 3u, 使用CAT法antibody screening negative, QD輸血1U
三天, 第一天輸血使用is交叉, 第二天則使用mp交叉, 但都相合血, 第三天mp交叉不合, 經抗體鑑定無
抗體, 重新發出血液
 經重新MP antibody screening 為2+,0,0
 MP抗體鑑定為MP only anti-E

37. 案例討論
 假如輸了MP-only抗體交叉不合的血, 輸血反應及輸注療效評估。
 MP only 抗體的來源可能性
- IgM抗體
- 之前輸血或懷孕產生，經長期未刺激維持低titer的存在
- 最近輸血產生，active IgM先產生或產生titer尚無法使用CAT或AHG偵測到，但MP法可以偵試到‧

38. Anti-Mia MP only trans-placenta
 A case report of a anti-Mia (MP only )
 Because of few fetus plasma , MP antibody screening positive, but CAT antibody screening
negative
 recheck mother antibody screening : CAT antibody screening negative, MP only anti-Mia antibody
present.
小張的報告 Page 38

39. Mother antibody screening ( MP only)
小張的報告 Page 39

40. Baby antibody screening ( MP only )
小張的報告 Page 40

41. Continue study
 MP antibody screening positive, CAT antibody identification negative, MP-only antibody.
 CAT antibody screening negative，從MP cross match , 可以找到交叉不相容的血液，進而鑑定出
MP-only allo-antibody.
 從CAT antibody screening positive , period transfusion , antibody decreasing 是否可以找到MP-only
antibody?
小張的報告 Page 41

42. CAT Antibody screening decreasing during period transfusion
Ab screen日期姓名抗體鑑定日期MP 備註
2014/7/22 邵華x -E 2010/8/26 1,0,0 CAT negative
2014/7/23 林麗x -Fyb 2014/4/10 0,0,0 CAT negative
2014/8/24 呂永x -c,-E,-S,-Wra 2013/7/18 1,0,0 CAT negative
2014/8/24 林梅x -c.-E 2012/8/2 1,0,0 CAT negative
2014/8/24 吳金x -E 2011/4/25 w,0,0 CAT negative
2014/9/24 蔣秀x -E 2013/4/24 0,0,0 CAT negative
2014/9/27 陳桂x -C,-e 2011/12/26 0,w,w CAT negative
2014/9/28 林于x -Mia 2013/9/29 0,0,0 CAT negative
2014/9/28 黃士x -Mia 2011/10/27 0,w,0 CAT negative
小張的報告 Page 42

43. 小張的報告 Page 43

44. 小張的報告 Page 44

45. 小張的報告 Page 45

46. 玻璃試管下顯微鏡觀看結果
小張的報告 Page 46

47. Outline
 Definition of clinical significant antibody & MP only allo-antibody
 Background
 Literature review
 Purpose
 Case reports
 Laboratory finding
 Conclusion
小張的報告 Page 47

48. Conclusion
 The MP only allo-antibody show
induced antibodies
due to DAT (polyAHG and mono-IgG) positive( DPC)
eluent specificity same as origin plasma
IgG or IgG+IgM form
same reacted with enzyme treated panel cells
 Need DAT IgG1/IgG3 confirm hemolysis potential.
 If gel or CAT automation antibody screen was done,
cross-matching with MP method needed
小張的報告 Page 48

49. Thanks for attention
張志昇
FB:張志昇
Email: jschang@seed.net.tw
Line: jschang
小張的報告 Page 49