详解抗凝收集血浆的方法

<p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">收集抗凝全血，轻轻颠倒充分抗凝后，可直接离心分离血浆待用或保存。根据不同抗凝剂的性能特征，选择合适的抗凝剂。实验室常用的抗凝剂有肝素的各种盐、</font>EDTA<font face="宋体">及枸橼酸钠。其方法如下：</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="background-color: rgb(204, 255, 204);"><span style="color: rgb(255, 102, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">1<font face="宋体">、肝素抗凝：</font></span></strong></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">最常用的抗凝剂，一般情况下对相关指标都不会有干扰，同时抗凝比例较大（抗凝剂：</font> <font face="宋体">全血</font>=1:30<font face="宋体">），对血液基本没有稀释影响。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">肝素是一种含有硫酸基团的粘多糖，带有强大的负电荷，具有加强抗凝血酶</font>III<font face="宋体">灭活丝 氨酸蛋白酶的作用，从而阻止凝血酶的形成，并有阻止血小板聚集等多种抗凝作用。肝素管一般用于生化及血流变的检测，是电解质检测的</font><font face="Calibri">zui</font><font face="宋体">佳选择，检验血标本中的钠离子时，不能使用肝素钠，以免影响检测结果。也不能用于白细胞计数和分类，因肝素会引起白细胞聚集。 </font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">尽管用肝素的三种盐抗凝所得电解质浓度无显著差别，但肝素锂还是被认为是</font>zui<font face="宋体">好的。这主要是人们认为肝素钠可使钠的测定值偏高，肝素铵可使血氨测定值偏高（尿素酶法测定）。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">肝素可抑制分子生物学中常用的一些工具酶，如限制性内切酶、</font>Taq<font face="宋体">酶等，可影响</font><font face="Calibri">PCR </font><font face="宋体">的实验结果。可采用一些方法消除肝素的抑制效应，如利用肝素酶，或在分离白细胞后用缓冲液洗涤白细胞两次以上等。然而，许多实验工作者仍不愿意在进行分子生物学实验时采用肝素作抗凝剂。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="background-color: rgb(204, 255, 204);"><span style="color: rgb(255, 102, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">2<font face="宋体">、</font><font face="Calibri">EDTA</font><font face="宋体">抗凝：</font></span></strong></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">乙二胺四乙酸是一种氨基多羧基酸，可以有效地鳌合血液中的钙离子，鳌合钙会将钙从</font> <font face="宋体">反应点移走将阻止和终止内源性或外源性凝血过程，从而防止血液凝固，与其他抗凝剂比较而言，其对血球的凝集及血细胞的形态影响较小，故通常使用</font>EDTA<font face="宋体">盐</font><font face="Calibri">(2K</font><font face="宋体">、</font><font face="Calibri">3K</font><font face="宋体">、</font><font face="Calibri">2Na)</font><font face="宋体">作为抗凝剂。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">用于一般血液学检查，不能用于血凝、微量元素及</font>PCR<font face="宋体">检查。由于</font><font face="Calibri">EDTA</font><font face="宋体">会螯合重金属离子，用该抗凝剂的话，部分带有金属离子的大分子酶都会有大量损失，具体看客户测定的指标来选择，做血常规的话必须用</font><font face="Calibri">EDTA</font><font face="宋体">抗凝。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="background-color: rgb(204, 255, 204);"><span style="color: rgb(255, 102, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">3<font face="宋体">、枸橼酸盐抗凝：</font></span></strong></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">柠檬酸钠通过作用于血样中钙离子鳌合而起抗凝作用，国家临床实验室标准化委员会</font>(NCCLS)<font face="宋体">推荐为</font><font face="Calibri">3.2</font><font face="宋体">％或</font><font face="Calibri">3.8</font><font face="宋体">％，抗凝剂与血比例为</font><font face="Calibri">1</font><font face="宋体">：</font><font face="Calibri">9</font><font face="宋体">，主要用于纤溶系统</font><font face="Calibri">(</font><font face="宋体">凝血酶原时间、凝血酶时间、活化部分凝血酶时间、纤维蛋白原</font><font face="Calibri">)</font><font face="宋体">。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">采血时应注意采足血量，以保证检验结果的准确性，采血后应立即轻轻颠倒混匀</font>5-8<font face="宋体">次。由于抗凝比例较小（抗凝剂：全血</font><font face="Calibri">=1:9</font><font face="宋体">），对血液有一定的稀释，一般不建议。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="font-family: 宋体; font-size: 9pt;">选择抗凝的注意点：</span></strong><span style="font-family: 宋体; font-size: 9pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">①&nbsp;</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">每一份样本所加的抗凝剂的量要一致，同时所取的全血的量也要尽量一致</font><font face="Calibri">;</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">②</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;<font face="宋体">收集抗凝全血后一定要轻轻颠倒，充分抗凝，防止部分血液未接触到抗凝剂而导致凝固</font><font face="Calibri">;</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">③</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;<font face="宋体">抗凝全血收集的血浆相对较多（</font><font face="Calibri">1ml</font><font face="宋体">抗凝全血能分离出</font><font face="Calibri">0.4-0.5ml</font><font face="宋体">血浆）</font><font face="Calibri">;</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">④</span><span style="font-size: 9pt;"><font face="Calibri">&nbsp;</font></span></strong><span style="font-size: 9pt;"><font face="宋体" style="font-family: 宋体;">抗凝收集的血浆冷冻保存后，解冻时可能会出现絮状浑浊，如有则需要离心去掉浑浊后 用于测定。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="color: rgb(255, 102, 0);"><span style="font-family: 宋体; font-size: 9pt;">&nbsp;</span></span></strong></p> <p class="MsoNormal"><span style="background-color: rgb(204, 255, 204);"><strong><span style="color: rgb(255, 102, 0);"><span style="font-family: 宋体; font-size: 9pt;">4<font face="宋体">、血清、血浆收集好后，暂时不测定，可立即低温冻存，温度越低越好，中间如不反复冻融，</font><font face="Calibri">-20</font><font face="宋体">℃以下可保存一个月</font><font face="Calibri">,-70</font><font face="宋体">℃以下可保存三个月。</font></span></span></strong></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>