鐘青,許業(yè)棟,吳永國(guó),劉文毅,肖靜,胡國(guó)剛

(松崗人民醫(yī)院檢驗(yàn)科,廣東深圳518105)

部分血小板參數(shù)缺失時(shí)小紅細(xì)胞/裂紅細(xì)胞對(duì)XS1000i血細(xì)胞分析儀血小板計(jì)數(shù)的影響

鐘青,許業(yè)棟,吳永國(guó),劉文毅,肖靜,胡國(guó)剛

(松崗人民醫(yī)院檢驗(yàn)科,廣東深圳518105)

目的探討XS1000i血細(xì)胞分析儀部分血小板參數(shù)缺失時(shí)小紅細(xì)胞/裂紅細(xì)胞對(duì)血小板計(jì)數(shù)的影響.方法對(duì)XS1000i血細(xì)胞分析儀分析后MCV<80fl且部分血小板參數(shù)缺失的標(biāo)本再使用PENTRA120Retic血細(xì)胞分析儀計(jì)數(shù)血小板、顯微鏡計(jì)數(shù)血小板及瑞氏染色鏡檢.結(jié)果PLT>100X109/L時(shí),XS1000i與PENTRA120Retic、顯微鏡計(jì)數(shù)血小板的差異有統(tǒng)計(jì)學(xué)意義(P<0.01),PLT<100X109/L時(shí),XS1000i與PENTRA120Retic、顯微鏡計(jì)數(shù)血小板的結(jié)果差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.01);當(dāng)MCV分別為70~80fl,60~70fl,<60fl時(shí),XS1000i均高于顯微鏡計(jì)數(shù)血小板的結(jié)果,差異有統(tǒng)計(jì)學(xué)意義(P<0.01).顯微鏡檢查結(jié)果發(fā)現(xiàn):101例標(biāo)本均檢出小紅細(xì)胞或(和)裂紅細(xì)胞,其中60例伴有大血小板或(和)巨血小板,4例伴有大血小板和血小板聚集.結(jié)論對(duì)于電阻抗血細(xì)胞分析儀,當(dāng)MCV<80fl且部分血小板參數(shù)缺失時(shí)的標(biāo)本應(yīng)用顯微鏡或ICSH推薦的參考方法計(jì)數(shù)血小板,同時(shí)染色鏡檢異常血細(xì)胞的形態(tài)、數(shù)量及分布.

血液分析儀;血小板計(jì)數(shù);小紅細(xì)胞;裂紅細(xì)胞;血小板參數(shù)

XS-1000i血細(xì)胞分析儀是Sysmex公司開發(fā)的一款體積最小的機(jī)型,可檢測(cè)WBC,RBC,HGB, HCT,MCV,MCH,MCHC,PLT,NEUT%,LYMPH%, MONO%,EO%,BASO%,NEUT#,LYMPH#,MONO#, EO#,BASO#,RDW-SD,RDW-CV,PDW,MPV,PCT,PLCR24個(gè)參數(shù),還提供IG%,IG#2個(gè)研究參數(shù),以及紅細(xì)胞直方圖、血小板直方圖、白細(xì)胞分類散點(diǎn)圖,極大地滿足了各種規(guī)模的醫(yī)院對(duì)臨床標(biāo)本檢測(cè)的需求.在實(shí)際運(yùn)用過(guò)程中,經(jīng)常遇到某些血液疾病如地中海貧血、缺鐵性貧血及某些慢性病貧血等標(biāo)本中的部分血小板參數(shù)如PDW(血小板體積分布寬度)、MPV(平均血小板體積)、PCT(血小板比容)、P-LCR(大型血小板比率)的檢測(cè)結(jié)果為"----",且血小板直方圖異常(如圖1,2),這給臨床疾病的診療和檢驗(yàn)報(bào)告的規(guī)范帶來(lái)諸多不利[1].筆者使用多種方法對(duì)該類標(biāo)本的血小板進(jìn)行檢測(cè),包括XS1000i、PENTRA120Retic、顯微鏡計(jì)數(shù)法、瑞氏染色鏡檢等,現(xiàn)將檢測(cè)結(jié)果報(bào)告如下.

圖1 血小板直方圖

圖2 血小板直方圖

1 材料與方法

1.1 儀器XS-1000i全自動(dòng)血液分析儀,由sysmex公司生產(chǎn);PENTRA120Retic全自動(dòng)血液分析儀,由HORIBA公司生產(chǎn);Olympus-CX31顯微鏡,由Olympus公司生產(chǎn);血球計(jì)數(shù)板,上海市求精生化試劑儀器有限公司生產(chǎn).

1.2 試劑XS-1000i血細(xì)胞分析儀原裝配套試劑及質(zhì)控品,PENTRA120Retic血細(xì)胞分析儀原裝配套試劑及質(zhì)控品,EDTA-K2抗凝管由廣州陽(yáng)普醫(yī)療科技股份有限公司提供,瑞氏染液由珠海BASO公司提供,血小板計(jì)數(shù)稀釋液(1%草酸銨稀釋液)按照《中華醫(yī)學(xué)檢驗(yàn)全書》[2]進(jìn)行配制.

1.3 標(biāo)本來(lái)源來(lái)自于2011年4月18日至2011年5月12日就診的門診及住院病人的靜脈血(n= 101),嚴(yán)格按照儀器原始操作手冊(cè)和《全國(guó)臨床檢驗(yàn)操作規(guī)程》[3]進(jìn)行真空采血管采集,標(biāo)本采集后顛倒混勻10次,肉眼可見標(biāo)本適量、無(wú)凝固、無(wú)凝塊、無(wú)溶血.

1.4 方法

1.4.1 嚴(yán)格按照血細(xì)胞分析儀的要求定期對(duì)XS-1000i血液分析儀、PENTRA120Retic血液分析儀進(jìn)行校準(zhǔn),每天儀器開機(jī)后進(jìn)行質(zhì)控檢測(cè),在各校準(zhǔn)參數(shù)和質(zhì)控參數(shù)均符合相關(guān)標(biāo)準(zhǔn)后,再對(duì)各標(biāo)本進(jìn)行自動(dòng)檢測(cè).

1.4.2 采集的靜脈血先用XS1000i血細(xì)胞分析儀進(jìn)行自動(dòng)檢測(cè),根據(jù)檢測(cè)結(jié)果,對(duì)MCV<80fl且無(wú)MPV、PCT、PDW、P-LCR參數(shù)的標(biāo)本先用PENTRA120Retic血液分析儀進(jìn)行自動(dòng)檢測(cè),然后用顯微鏡計(jì)數(shù)法計(jì)數(shù)血小板,同時(shí)進(jìn)行血涂片制備和染色,標(biāo)本采集后在4小時(shí)內(nèi)完成以上檢測(cè).

1.4.3 經(jīng)嚴(yán)格培訓(xùn)且有理論、實(shí)踐經(jīng)驗(yàn)的檢驗(yàn)人員進(jìn)行細(xì)胞形態(tài)學(xué)檢查選擇理想的區(qū)域進(jìn)行鏡檢(理想紅細(xì)胞分布區(qū)域是紅細(xì)胞之間相近排列而不重疊),用油鏡觀察血膜體尾交界處的細(xì)胞形態(tài)[4].小紅細(xì)胞/裂紅細(xì)胞>2%有診斷價(jià)值[5](注:小紅細(xì)胞:直徑小于6μm,大血小板:直徑5~7.5μm,巨血小板:直徑大于7.5μm[6].)

1.5 分組

1.5.1 按XS1000i血細(xì)胞分析儀血小板的檢測(cè)結(jié)果分為高、低2組,高值組(n=98):PLT>100X109/L,低值組(n=3):PLT<100X109/L;

1.5.2 按XS1000i血細(xì)胞分析儀MCV的檢測(cè)結(jié)果分為3組:(a)MCV:70~80fl,n=21,(b)MCV:60~70fl, n=59,(c)MCV:<60fl,n=21

1.6 統(tǒng)計(jì):使用SPSS Statistics 17.0統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì),進(jìn)行配對(duì)樣本t檢驗(yàn),數(shù)據(jù)以均值±標(biāo)準(zhǔn)差()表示,P<0.01表示差異有統(tǒng)計(jì)學(xué)意義.

2 結(jié)果

2.1 XS1000i與顯微鏡計(jì)數(shù)法對(duì)血小板計(jì)數(shù)結(jié)果的比較,XS1000i與PENTRA120Retic對(duì)血小板計(jì)數(shù)結(jié)果的比較分別見表1和表2.從表1可以看到,高值組XS1000i比顯微鏡計(jì)數(shù)法的結(jié)果要偏高,相關(guān)系數(shù)為0.749,P=0.000,差異有統(tǒng)計(jì)學(xué)意義,而低值組XS1000i與顯微鏡計(jì)數(shù)法檢測(cè)的PLT結(jié)果接近,相關(guān)系數(shù)為0.988,P=0.099,差異無(wú)統(tǒng)計(jì)學(xué)意義,可能與標(biāo)本量(n=3)過(guò)少有關(guān).

表1 XS1000i與顯微鏡計(jì)數(shù)法血小板計(jì)數(shù)結(jié)果的比較

表2 XS1000i與PENTRA120Retic血小板計(jì)數(shù)結(jié)果的比較

從表2可以看到,高值組XS1000i比PENTRA120Retic的結(jié)果偏高,相關(guān)系數(shù)為0.858,P= 0.000,差異有統(tǒng)計(jì)學(xué)意義,而低值組XS1000i與PENTRA120Retic檢測(cè)的PLT結(jié)果相近,相關(guān)系數(shù)為0.962,P=0.176,差異無(wú)統(tǒng)計(jì)學(xué)意義,也可能與標(biāo)本量(n=3)過(guò)少有關(guān).

2.2 XS1000i與顯微鏡計(jì)數(shù)法對(duì)MCV不同標(biāo)本的血小板計(jì)數(shù)結(jié)果比較見表3.從表3可以看到,各組的血小板檢測(cè)結(jié)果均是XS1000i比顯微鏡計(jì)數(shù)法高,P值均<0.01,差異有統(tǒng)計(jì)學(xué)意義.

表3 XS1000i與顯微鏡計(jì)數(shù)法對(duì)MCV不同標(biāo)本的血小板計(jì)數(shù)結(jié)果比較

2.3 顯微鏡檢查結(jié)果101例標(biāo)本均檢出小紅細(xì)胞或(和)裂紅細(xì)胞,其中60例伴有大血小板或(和)巨大血小板(如圖3),4例伴有大血小板和血小板聚集,其余37例標(biāo)本未見血小板異常.

圖3 外周血裂紅細(xì)胞、大血小板和巨血小板形態(tài)(X1000)

3 討論

XS1000i血細(xì)胞分析儀是利用流體聚焦法分析紅細(xì)胞和血小板計(jì)數(shù),使用3個(gè)界標(biāo)(2~6fl,12fl, 12~30fl)檢測(cè)血小板的顆粒大小分布,改善了血細(xì)胞計(jì)數(shù)的精確度和重現(xiàn)性,防止產(chǎn)生假性血小板脈沖.但出現(xiàn)小于30fl的小紅細(xì)胞或細(xì)胞碎片時(shí),這些小紅細(xì)胞和碎片會(huì)當(dāng)成血小板計(jì)數(shù),導(dǎo)致血小板假性偏高.

小紅細(xì)胞(MCV<80fl)一般出現(xiàn)在缺鐵性貧血、鐵粒幼細(xì)胞貧血、血紅蛋白病(如:珠蛋白合成障礙性貧血)、遺傳性球形細(xì)胞增高癥及慢性疾病所引起的貧血中,而這些增生性貧血易出現(xiàn)紅細(xì)胞碎片,因此極易干擾PLT的檢測(cè)[7].一般裂紅細(xì)胞增多見于微血管病性溶血性貧血、血栓性血小板減少性紫癜、溶血尿毒癥綜合征、惡性高血壓、創(chuàng)傷性心血管性溶血性貧血等[8].

從表1和表2可以看出,當(dāng)血小板減少時(shí),兩臺(tái)血細(xì)胞分析儀之間以及血細(xì)胞分析儀與顯微鏡計(jì)數(shù)法計(jì)數(shù)血小板的差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.01),相關(guān)系數(shù)分別是0.962、0.988,顯示了良好的相關(guān)性,小紅細(xì)胞/裂紅細(xì)胞對(duì)血小板計(jì)數(shù)無(wú)明顯影響,可能與本次檢測(cè)標(biāo)本量過(guò)少有關(guān).Trabuio等使用多種方法對(duì)原發(fā)性血小板減少性紫癜的血小板計(jì)數(shù)(PLT<20X109/L)進(jìn)行比較顯示了良好的相關(guān)性[9],與本文結(jié)果一致.當(dāng)血小板>100X109/L時(shí),兩臺(tái)血細(xì)胞分析儀之間以及血細(xì)胞分析儀與顯微鏡計(jì)數(shù)法計(jì)數(shù)血小板的相關(guān)系數(shù)分別是0.858、0.749,P值均為0.000(P<0.01),差異有統(tǒng)計(jì)學(xué)意義,小紅細(xì)胞、裂紅細(xì)胞等對(duì)血小板計(jì)數(shù)有明顯的影響.

從表3可以看到,各組的血小板檢測(cè)結(jié)果均是XS1000i比顯微鏡計(jì)數(shù)法高,P<0.01,,差異有統(tǒng)計(jì)學(xué)意義,與梁瑞蓮[10]季明德[11]、張霞等[12]的研究結(jié)果一致.所有101例標(biāo)本經(jīng)顯微鏡涂片檢查均檢出小紅細(xì)胞或(和)裂紅細(xì)胞,部分標(biāo)本還伴有大血小板/巨血小板/血小板聚集現(xiàn)象等等.因此在使用阻抗法檢測(cè)血小板/紅細(xì)胞的血細(xì)胞分析儀如XS1000i、PENTRA120Retic時(shí),若部分血小板參數(shù)缺失,應(yīng)注意紅細(xì)胞碎片、有核細(xì)胞的細(xì)胞質(zhì)碎片、冷球蛋白、細(xì)菌、真菌、油脂等[13]的脈沖信號(hào)可能視為血小板信號(hào)而干擾實(shí)驗(yàn)結(jié)果,造成實(shí)驗(yàn)誤差,導(dǎo)致血小板的假性升高[14,15],這時(shí)要進(jìn)行顯微鏡計(jì)數(shù)法計(jì)數(shù)血小板或使用ICSH推薦的參考方法計(jì)數(shù)血小板,同時(shí)進(jìn)行血涂片檢查并在檢驗(yàn)報(bào)告單上注明異常細(xì)胞的形態(tài)、數(shù)量和分布等等.

[1]閔迅,陳麗華,駱科允.XT-1800i血細(xì)胞分析儀不能提供完整血小板參數(shù)的原因及對(duì)策[J].現(xiàn)代檢驗(yàn)醫(yī)學(xué)雜志,2008,23:38.

[2]李影林.中華醫(yī)學(xué)檢驗(yàn)全書[M].北京:人民衛(wèi)生出版社,2000:202.

[3]葉應(yīng)嫵,王毓三,申子瑜.全國(guó)臨床檢驗(yàn)操作規(guī)程[M].第3版.南京:東南大學(xué)出版社,2006:121-122.

[4]叢玉隆,尹一兵,陳瑜.檢驗(yàn)醫(yī)學(xué)高級(jí)教程[M].北京:人民軍醫(yī)出版社,2010:108.

[5]王永才.血液骨髓細(xì)胞檢驗(yàn)診斷學(xué)[M].大連:大連出版社,1995:21-22.

[6]劉志潔,黃文源,劉方文.臨床血液學(xué)及細(xì)胞學(xué)圖譜[M].第3版.北京:科學(xué)出版社,2006:70-83.

[7]鄒漢良,祝玲玲,蔣明,等.XS-800i血細(xì)胞分析儀對(duì)地中海貧血患者血小板參數(shù)檢測(cè)的影響分析[J].國(guó)際檢驗(yàn)醫(yī)學(xué)雜志,2009,30 (7):639-640.

[8]王鴻利.實(shí)驗(yàn)診斷學(xué)[M].北京:人民衛(wèi)生出版社,2005:35.

[9]Trabuio E,Valverde S.Antico F,et al.Performance of automated platelet quantification using different analysers in comparison with an immunological reference method in thrombocytopenic patients [J].Blood Transfus,2009,7(1):43-48.

[10]梁瑞蓮,周遠(yuǎn)青,唐躍華,等.小紅細(xì)胞對(duì)不同原理血液分析儀的血小板計(jì)數(shù)的影響[J].國(guó)際檢驗(yàn)醫(yī)學(xué)雜志,2009,30(7):699.

[11]季明德,顧萬(wàn)健.Sysmex XT-2000i檢測(cè)血小板直方圖分布異常時(shí)儀器計(jì)數(shù)與手工計(jì)數(shù)的比較[J].醫(yī)學(xué)檢驗(yàn)與臨床,2007,18:69.

[12]張霞.影響Sysmex-1800i血液分析儀血小板計(jì)數(shù)因素復(fù)檢原則的探討[J].中國(guó)醫(yī)藥導(dǎo)報(bào),2010,7(15):73

[13]Zandecki M,Genevieve F,Gerard J.Spurious counts and spurious results on haematology analysers:a review.Part I:platelets[J].Int J Lab Hematol,2007,29:4-20.

[14]彭文紅,樂(lè)家新,蘭亞婷,等."微孔故障"影響血小板計(jì)數(shù)及直方圖變化的探討[J].陜西醫(yī)學(xué)檢驗(yàn),2001,16(3):30-31.

[15]Tatsumi N,Tsuda I,Funahara Y.Assessment of hematological data obtained by the aperture impedance method in blood from patients with thalassemia or hemoglobinpoathy[J].ICMR Annals,1998,8(1): 49-60.

Effect of microcytes/schistocytes on platelet count when deficiency of portion platelet parameters measured by XS1000i Hematology analyzer

ZHONG Qing,XU Yedong,WU Yongguo,et al.Department of Laboratory Medicine,Songgang People's Hospital,Shenzhen Guangdong 518105,China

ObjectiveTo investigate the effect of microcytes/schistocytes on platelet count when deficiency of portion platelet parameters measured by XS1000i hematology analyzer.MethodsAfter analyzed by XS1000i hematology analyzer,the samples which have MCV<80fl and loss partial platelet parameters were re-measured with the PENTRA120Retic hematology analyzer,microscopic platelet count and,Wright staining for platelets.ResultsWhen PLT>100X109/L,there were statistically significant differences in the platelet counts among XS1000i,PENTRA120Retic and microscopic counting(P<0.01);when PLT<100X109/L,there were no statistically significant differences in the platelet counts among XS1000i PENTRA120Retic and microscopic counting(P> 0.01).When MCV was 70~80fl,60~70fl and<60fl,respectively,the platelet counts of XS1000i were higher than those of microscopic counting,the difference was statistically significant(P<0.01).The results of microscope examination were as follows:all 101 specimens were detected and had microcytes or(and)schistocytes,of which 60 cases were accompanied with large platelets and/or giant platelets,and 4 cases were accompanied with large platelets and platelet aggregation.ConclusionsThe samples with MCV< 80fl and missing partial platelet parameters when analyzed by impedance hematology analyzer should be re-measured with the reference method recommended by ICSH or microscopic platelet counting,meanwhile,detect the morphology,number and distribution of abnormal blood cells by staining and microscopic examination.

Hematology analyzer;Platelet count;Microcyte;Schistocyte;Platelet parameters

R446.11+1

A

1674-1129(2014)03-0264-03

10.3969/j.issn.1674-1129.2014.03.011

2013-10-29;

2014-2-25)

鐘青,女,1970年出生,主管技師,主要研究方向?yàn)榕R床血液學(xué).

許業(yè)棟,男,1967年出生,副主任技師.