Mp only 馬偕

Are manual Polybrene ( MP) only antibodies clinical

significance ? Serological View.

張志昇

馬偕紀念醫院

2014/11/5

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 2

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


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）．


Outline


Background

Literature review

Purpose

Case reports

Laboratory finding

Conclusion


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。


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

Reference

Conclusions

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。

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。

ObjectivesMP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%

Table 1.MP & CAT檢出抗體陽性結果比較表

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%)

Table 2.MP & CAT檢出抗體特異性結果分析表

冷型抗體


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%)

AHG抗體


Specificity

Antibody frequency

Jka 9 (1.8%) 14 (2.7%)

Jkb

Jk34 (0.8%)1 (0.2%)

5 (0.96%)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

Anti-Lea

Figure 3. CAT法檢出抗體

Anti-Mia

Anti-E & c

Anti-M

Manual polybrene Ab SRC (AC not performed)

CAT panel & AC

CAT panel & AC negative

Repeat MP Ab SRC

No further

MP Ab panel

IS XMDetermine 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

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

ABScr 3 Poly

AutoVue-5065 (AHG IgG) Autovue-5066 (AHG IgG) AutoVue-原機 (AHG POLY) MP法

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



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



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

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！

結論

40400430400430400A6

+/-1400330400330400A5

00044000?MF44FIB0?MF4A4

30400200400200400A3

004100004?00004?0A2

33100430000430000A1

B cellsA1 cellsAnit-DAnti-BAnti-AB cellsA1 cellsControlAnit-DAnti-BAnti-AB cellsA1 cellsControlAnit-DAnti-BAnti-Asample

tube methodAutoVue (5066)AutoVue (5065)

024未測

0345NEG

2018POS

AutoVue (7272)Poly

0012未測

0351NEG

2114POS

AutoVue (5066)IgG

000未測

0357NEG

2120POS

AutoVue (5065)IgG

未測NEGPOS

MP method抗體篩檢結果比較

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tubemethod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。

2.抗體篩檢在MP法與AutoVue之間的比對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 card做65個檢體比對。36個檢體在MP法表現與AutoVue 兩種卡片篩檢皆為Negative(僅sample 2 之MP法表現為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

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。

簡介

輸血前檢驗，包括ABO/RH血型、不規則抗體篩檢、抗體鑑定

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。

本院以AutoVue自動化血型鑑定之柱狀凝集法CAT(column 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 Type1. Tube Method:• Anti-A & Anti-B: NOVACLONE, Murine Monoclonal• Anti-D: NOVACLONE, Anti-D IgM+IgGMonoclonal 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 Screen1. Manual Polybrene(MP):

0.8% cells & Serum

Air Gap

Antihuman Globulin

and Diluent

Glass Beads



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%

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


RH類及其他類抗體



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



Jka 9 (1.8%) 14 (2.7%)

Jkb

Jk34 (0.8%)1 (0.2%)

5 (0.96%)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



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%)

Outline


Background

Literature review

Purpose

Case reports

Laboratory finding

Conclusion


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.


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


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


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

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.

Evaluation of Ortho BioVue system, the DiaMed Gel system, and Gamma-

ReACT test and their comparison with Manual Polybrene test (II)


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


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

antibodies

0 6 3 10 3

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)。

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

MP sensitive

2014/11/422

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

Outline


Background

Literature review

Purpose

Case reports

Laboratory finding

Conclusion


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。


Outline


Background

Literature review

Purpose

Case reports

Laboratory finding

Conclusion


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.


Outline


Background

Literature review

Purpose

Case reports

Laboratory finding

Conclusion



Result of manual Polybrene and Rh phenotype


Manual Polybrene Test ( MPT-AGT )


0.02M DTT (DL-DITHIOTHREITOL) treated


Enzyme treated panel cells reactive @ RT with poly AHG card


Enzyme treated panel cell @ 37°C with mono-IgG card


Direct Coombs’ test ( microscope view)


Eluent from DAT positive cells


仍具anti-E, anti-Jka特性, 唯anti-E 仍呈現 MP only 特性。

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

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‧

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.


Mother antibody screening ( MP only)


Baby antibody screening ( MP only )


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?


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





玻璃試管下顯微鏡觀看結果


Outline


Background

Literature review

Purpose

Case reports

Laboratory finding

Conclusion


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


Thanks for attention張志昇

FB:張志昇

Email: [email protected]

Line: jschang


mailto:[email protected]

Health & Medicine

Mp only 馬偕