探讨预固定对中性粒细胞碱性磷酸酶染色结果的影响
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摘要
目的探讨预固定对中性粒细胞碱性磷酸酶(NAP)的阳性率和积分产生的影响。方法随机抽取182例来某自三甲综合医院血液科住院患者的新鲜静脉血,乙二胺四乙酸二钾(EDTA-K2)真空管抗凝,制备3份血涂片,第1份(A)在1h内进行NAP染色,第2份(B)预先进行固定,1d后进行NAP染色,第3份(C)不做任何处理,1d后进行NAP染色;同时在其做骨穿时取其新鲜末梢血,制备血涂片,在1h内进行NAP染色;将上述血涂片用NAP染色液进行NAP细胞组化染色,用显微镜在油镜视野下观察100个中性杆状、分叶核粒细胞,以阳性率和积分记录其检测结果。结果 EDTA-K2抗凝静脉血涂片A的NAP阳性率和积分与新鲜末梢血相比差异无统计学意义(P>0.05);EDTA-K2抗凝静脉血涂片B的NAP阳性率和积分与新鲜末梢血相比略低,但差异仍无统计学意义(P>0.05);EDTA-K2抗凝静脉血涂片C的NAP阳性率和积分与新鲜末梢血相比差异有统计学意义(P<0.05)。结论经过预固定的EDTA-K2抗凝静脉血涂片放置1d后,其NAP染色的结果依然可靠,不经过固定的EDTA-K2抗凝静脉血涂片放置1d后,其NAP染色的结果显著降低。
Objective To explore the influence of fixed in advance on the positive rate and integral of neutrophilic alkaline phosphatase(NAP)dyeing.Methods Totally 182 cases of fresh venous blood from inpatients in the top three hospital department of hematology were randomly selected and anticoagulated in the EDTA-K2 vacuum tube.Three blood smears from which were prepared as follows:the first blood smear(Named A)NAP dyeing completed within an hour;the second one(Named B)were fixed in advanceand NAP dyeing after one day;the third one(Named C)didn′t do any processing and NAP dyeing after one day.At the same time,the blood samples were taken from the fresh blood,and the blood smears were prepared and stained with NAP in an hour.NAP dyeing were performed by NAP dyeing for the blood smears,and 100 neutral rods,nucleus granulocyte were observed under microscope in the oil mirror vision,the test results record by positive rate and integral.Results No significant difference of NAP positive rate and integral was found in EDTA-K2 anticoagulation venous blood smear A when compared with fresh peripheral blood(P>0.05).The NAP positive rate and integral of EDTA-K2 anticoagulation venous blood smear B was slightly lower than that from fresh peripheral blood,but no significant difference was found(P>0.05).However,the NAP positive rate and integral in EDTA-K2 anticoagulant venous blood smear C has a significant difference from fresh peripheral blood(P<0.05).Conclusion The NAP dyeing results of EDTA-K2 anticoagulation venous blood smear fixed and placed one days are still reliable,while the NAP dyeing results was significantly reduced in the unfixed EDTA-K2 anticoagulation venous blood smear placed after one day.
Objective To explore the influence of fixed in advance on the positive rate and integral of neutrophilic alkaline phosphatase(NAP)dyeing.Methods Totally 182 cases of fresh venous blood from inpatients in the top three hospital department of hematology were randomly selected and anticoagulated in the EDTA-K2 vacuum tube.Three blood smears from which were prepared as follows:the first blood smear(Named A)NAP dyeing completed within an hour;the second one(Named B)were fixed in advanceand NAP dyeing after one day;the third one(Named C)didn′t do any processing and NAP dyeing after one day.At the same time,the blood samples were taken from the fresh blood,and the blood smears were prepared and stained with NAP in an hour.NAP dyeing were performed by NAP dyeing for the blood smears,and 100 neutral rods,nucleus granulocyte were observed under microscope in the oil mirror vision,the test results record by positive rate and integral.Results No significant difference of NAP positive rate and integral was found in EDTA-K2 anticoagulation venous blood smear A when compared with fresh peripheral blood(P>0.05).The NAP positive rate and integral of EDTA-K2 anticoagulation venous blood smear B was slightly lower than that from fresh peripheral blood,but no significant difference was found(P>0.05).However,the NAP positive rate and integral in EDTA-K2 anticoagulant venous blood smear C has a significant difference from fresh peripheral blood(P<0.05).Conclusion The NAP dyeing results of EDTA-K2 anticoagulation venous blood smear fixed and placed one days are still reliable,while the NAP dyeing results was significantly reduced in the unfixed EDTA-K2 anticoagulation venous blood smear placed after one day.
引文
[1]许文荣.临床血液学和血液检验实验指导[M].2版.北京:人民卫生出版社,2006.
[2]叶应妩,王毓三.全国临床检验操作规程[M].2版.南京:东南大学出版社,1997.
[3]邓宽国.三种抗凝方法对中性粒细胞碱性磷酸酶染色结果的分析[J].医学检验与临床,2005,16(2):64.
[4]张立文,冉宝春,王海,等.成人中性粒细胞碱性磷酸酶影响因素探讨[J].解放军医学院学报,2004,25(2):152-153.
[5]王鸿利.实验诊断学[M].北京:人民卫生出版社,2001.
[6]Borregaard N,Kjeldsen L,Lollike K,et al.Granules and secretory vesicles of the human neutrophil[J].Clin Exp Immunol,1995,101(s1):6-9.
[7]Rustin GJ,Wilson PD,Peters TJ.Studies on the subcellular localization of human neutrophil alkaline phosphatase[J].J Cell Sci,1979,36(4):401-412.
[8]Smith GP,Sharp G,Peters TJ.Isolation and characterization of alkaline phosphatase-containing granules(phosphasomes)from human polymorphonuclear leucocytes[J].J Cell Sci,1985,76(10):167-178.
[9]许文荣,王建中.临床血液学与检验[M].4版.北京:人民卫生出版社,2010.
[10]Iqbal SJ,Davies T,Cole R,et a1.Neutrophil alkaline phosphatase(NAP)score in the diagnosis of hypophosphatasia[J].Clin Chim Aeta,2000,302(1/2):49-57.
[11]常玉荣,石峻,殷华,等.血细胞形态学观察对血液系统疾病诊断的意义[J].检验医学,2007,22(1):78-80.
[12]沈亚文,王仲泉,邓宽国.标本采集方法对中性粒细胞碱性磷酸酶染色结果影响的观察[J].临床和实验医学杂志,2003,2(2):126-127.
[13]许宏敏,顾春刚,周淑芬,等.抗凝血对中性粒细胞碱性磷酸酶染色结果的影响[J].广东医学,2012,33(12):1735-1736.
[14]郑丽梅,徐国洁,葛晓军.rhG-CSF对急性粒细胞白血病化疗后粒细胞缺乏患者NAP活性的影响[J].实验与检验医学,2011,29(1):47-48.
[15]余安乐.骨碱性磷酸酶免疫试剂盒在儿童缺钙检测中的应用[J].实验与检验医学,2008,26(6):683.
[16]汤萌,周义,田露.80例外周血中性粒细胞碱性磷酸酶染色与结果分析[J].实验与检验医学,2011,29(3):311.
[2]叶应妩,王毓三.全国临床检验操作规程[M].2版.南京:东南大学出版社,1997.
[3]邓宽国.三种抗凝方法对中性粒细胞碱性磷酸酶染色结果的分析[J].医学检验与临床,2005,16(2):64.
[4]张立文,冉宝春,王海,等.成人中性粒细胞碱性磷酸酶影响因素探讨[J].解放军医学院学报,2004,25(2):152-153.
[5]王鸿利.实验诊断学[M].北京:人民卫生出版社,2001.
[6]Borregaard N,Kjeldsen L,Lollike K,et al.Granules and secretory vesicles of the human neutrophil[J].Clin Exp Immunol,1995,101(s1):6-9.
[7]Rustin GJ,Wilson PD,Peters TJ.Studies on the subcellular localization of human neutrophil alkaline phosphatase[J].J Cell Sci,1979,36(4):401-412.
[8]Smith GP,Sharp G,Peters TJ.Isolation and characterization of alkaline phosphatase-containing granules(phosphasomes)from human polymorphonuclear leucocytes[J].J Cell Sci,1985,76(10):167-178.
[9]许文荣,王建中.临床血液学与检验[M].4版.北京:人民卫生出版社,2010.
[10]Iqbal SJ,Davies T,Cole R,et a1.Neutrophil alkaline phosphatase(NAP)score in the diagnosis of hypophosphatasia[J].Clin Chim Aeta,2000,302(1/2):49-57.
[11]常玉荣,石峻,殷华,等.血细胞形态学观察对血液系统疾病诊断的意义[J].检验医学,2007,22(1):78-80.
[12]沈亚文,王仲泉,邓宽国.标本采集方法对中性粒细胞碱性磷酸酶染色结果影响的观察[J].临床和实验医学杂志,2003,2(2):126-127.
[13]许宏敏,顾春刚,周淑芬,等.抗凝血对中性粒细胞碱性磷酸酶染色结果的影响[J].广东医学,2012,33(12):1735-1736.
[14]郑丽梅,徐国洁,葛晓军.rhG-CSF对急性粒细胞白血病化疗后粒细胞缺乏患者NAP活性的影响[J].实验与检验医学,2011,29(1):47-48.
[15]余安乐.骨碱性磷酸酶免疫试剂盒在儿童缺钙检测中的应用[J].实验与检验医学,2008,26(6):683.
[16]汤萌,周义,田露.80例外周血中性粒细胞碱性磷酸酶染色与结果分析[J].实验与检验医学,2011,29(3):311.