詳解抗凝收集血漿的方法

<p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;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="宋體">收集抗凝全血，輕輕顛倒充分抗凝后，可直接離心分離血漿待用或保存。根據(jù)不同抗凝劑的性能特征，選擇合適的抗凝劑。實(shí)驗(yàn)室常用的抗凝劑有肝素的各種鹽、</font>EDTA<font face="宋體">及枸櫞酸鈉。其方法如下：</font></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">1<font face="宋體">、肝素抗凝：</font></span></strong></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">最常用的抗凝劑，一般情況下對(duì)相關(guān)指標(biāo)都不會(huì)有干擾，同時(shí)抗凝比例較大（抗凝劑：</font> <font face="宋體">全血</font>=1:30<font face="宋體">），對(duì)血液基本沒(méi)有稀釋影響。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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:宋體;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="宋體">肝素是一種含有硫酸基團(tuán)的粘多糖，帶有強(qiáng)大的負(fù)電荷，具有加強(qiáng)抗凝血酶</font>III<font face="宋體">滅活絲 氨酸蛋白酶的作用，從而阻止凝血酶的形成，并有阻止血小板聚集等多種抗凝作用。肝素管一般用于生化及血流變的檢測(cè)，是電解質(zhì)檢測(cè)的</font><font face="Calibri">zui</font><font face="宋體">佳選擇，檢驗(yàn)血標(biāo)本中的鈉離子時(shí)，不能使用肝素鈉，以免影響檢測(cè)結(jié)果。也不能用于白細(xì)胞計(jì)數(shù)和分類，因肝素會(huì)引起白細(xì)胞聚集。 </font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">盡管用肝素的三種鹽抗凝所得電解質(zhì)濃度無(wú)顯著差別，但肝素鋰還是被認(rèn)為是</font>zui<font face="宋體">好的。這主要是人們認(rèn)為肝素鈉可使鈉的測(cè)定值偏高，肝素銨可使血氨測(cè)定值偏高（尿素酶法測(cè)定）。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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:宋體;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="宋體">肝素可抑制分子生物學(xué)中常用的一些工具酶，如限制性內(nèi)切酶、</font>Taq<font face="宋體">酶等，可影響</font><font face="Calibri">PCR </font><font face="宋體">的實(shí)驗(yàn)結(jié)果。可采用一些方法消除肝素的抑制效應(yīng)，如利用肝素酶，或在分離白細(xì)胞后用緩沖液洗滌白細(xì)胞兩次以上等。然而，許多實(shí)驗(yàn)工作者仍不愿意在進(jìn)行分子生物學(xué)實(shí)驗(yàn)時(shí)采用肝素作抗凝劑。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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: 宋體; font-size: 9pt;">2<font face="宋體">、</font><font face="Calibri">EDTA</font><font face="宋體">抗凝：</font></span></strong></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">乙二胺四乙酸是一種氨基多羧基酸，可以有效地鰲合血液中的鈣離子，鰲合鈣會(huì)將鈣從</font> <font face="宋體">反應(yīng)點(diǎn)移走將阻止和終止內(nèi)源性或外源性凝血過(guò)程，從而防止血液凝固，與其他抗凝劑比較而言，其對(duì)血球的凝集及血細(xì)胞的形態(tài)影響較小，故通常使用</font>EDTA<font face="宋體">鹽</font><font face="Calibri">(2K</font><font face="宋體">、</font><font face="Calibri">3K</font><font face="宋體">、</font><font face="Calibri">2Na)</font><font face="宋體">作為抗凝劑。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">用于一般血液學(xué)檢查，不能用于血凝、微量元素及</font>PCR<font face="宋體">檢查。由于</font><font face="Calibri">EDTA</font><font face="宋體">會(huì)螯合重金屬離子，用該抗凝劑的話，部分帶有金屬離子的大分子酶都會(huì)有大量損失，具體看客戶測(cè)定的指標(biāo)來(lái)選擇，做血常規(guī)的話必須用</font><font face="Calibri">EDTA</font><font face="宋體">抗凝。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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: 宋體; font-size: 9pt;">3<font face="宋體">、枸櫞酸鹽抗凝：</font></span></strong></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">檸檬酸鈉通過(guò)作用于血樣中鈣離子鰲合而起抗凝作用，國(guó)家臨床實(shí)驗(yàn)室標(biāo)準(zhǔn)化委員會(huì)</font>(NCCLS)<font face="宋體">推薦為</font><font face="Calibri">3.2</font><font face="宋體">％或</font><font face="Calibri">3.8</font><font face="宋體">％，抗凝劑與血比例為</font><font face="Calibri">1</font><font face="宋體">：</font><font face="Calibri">9</font><font face="宋體">，主要用于纖溶系統(tǒng)</font><font face="Calibri">(</font><font face="宋體">凝血酶原時(shí)間、凝血酶時(shí)間、活化部分凝血酶時(shí)間、纖維蛋白原</font><font face="Calibri">)</font><font face="宋體">。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">采血時(shí)應(yīng)注意采足血量，以保證檢驗(yàn)結(jié)果的準(zhǔn)確性，采血后應(yīng)立即輕輕顛倒混勻</font>5-8<font face="宋體">次。由于抗凝比例較小（抗凝劑：全血</font><font face="Calibri">=1:9</font><font face="宋體">），對(duì)血液有一定的稀釋，一般不建議。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">選擇抗凝的注意點(diǎn)：</span></strong><span style="font-family: 宋體; font-size: 9pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">每一份樣本所加的抗凝劑的量要一致，同時(shí)所取的全血的量也要盡量一致</font><font face="Calibri">;</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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:宋體;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="宋體">收集抗凝全血后一定要輕輕顛倒，充分抗凝，防止部分血液未接觸到抗凝劑而導(dǎo)致凝固</font><font face="Calibri">;</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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:宋體;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="宋體">抗凝全血收集的血漿相對(duì)較多（</font><font face="Calibri">1ml</font><font face="宋體">抗凝全血能分離出</font><font face="Calibri">0.4-0.5ml</font><font face="宋體">血漿）</font><font face="Calibri">;</font></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體" style="font-family: 宋體;">抗凝收集的血漿冷凍保存后，解凍時(shí)可能會(huì)出現(xiàn)絮狀渾濁，如有則需要離心去掉渾濁后 用于測(cè)定。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; 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: 宋體; font-size: 9pt;">4<font face="宋體">、血清、血漿收集好后，暫時(shí)不測(cè)定，可立即低溫凍存，溫度越低越好，中間如不反復(fù)凍融，</font><font face="Calibri">-20</font><font face="宋體">℃以下可保存一個(gè)月</font><font face="Calibri">,-70</font><font face="宋體">℃以下可保存三個(gè)月。</font></span></span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;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>