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。實驗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,一般情況下對相關(guān)指標(biāo)都不會有干擾,同時抗凝比例較大(抗凝劑:</font> <font face="宋體">全血</font>=1:30<font face="宋體">),對血液基本沒有稀釋影響。</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;"> </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的粘多糖,帶有強大的負電荷,具有加強抗凝血酶</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,是電解質(zhì)檢測的</font><font face="Calibri">zui</font><font face="宋體">佳選擇,檢驗血標(biāo)本中的鈉離子時,不能使用肝素鈉,以免影響檢測結(jié)果。也不能用于白細胞計數(shù)和分類,因肝素會引起白細胞聚集。 </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)濃度無顯著差別,但肝素鋰還是被認為是</font>zui<font face="宋體">好的。這主要是人們認為肝素鈉可使鈉的測定值偏高,肝素銨可使血氨測定值偏高(尿素酶法測定)。</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;"> </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">的實驗結(jié)果??刹捎靡恍┓椒ㄏ嗡氐囊种菩?yīng),如利用肝素酶,或在分離白細胞后用緩沖液洗滌白細胞兩次以上等。然而,許多實驗工作者仍不愿意在進行分子生物學(xué)實驗時采用肝素作抗凝劑。</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;"> </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,鰲合鈣會將鈣從</font> <font face="宋體">反應(yīng)點移走將阻止和終止內(nèi)源性或外源性凝血過程,從而防止血液凝固,與其他抗凝劑比較而言,其對血球的凝集及血細胞的形態(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有大量損失,具體看客戶測定的指標(biāo)來選擇,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;"> </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">檸檬酸鈉通過作用于血樣中鈣離子鰲合而起抗凝作用,國家臨床實驗室標(biāo)準(zhǔn)化委員會</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">凝血酶原時間、凝血酶時間、活化部分凝血酶時間、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">采血時應(yīng)注意采足血量,以保證檢驗結(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?。鼓齽喝?lt;/font><font face="Calibri">=1:9</font><font face="宋體">),對血液有一定的稀釋,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:</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 </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;"> <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;"> <font face="宋體">抗凝全血收集的血漿相對較多(</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"> </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,解凍時可能會出現(xiàn)絮狀渾濁,如有則需要離心去掉渾濁后 用于測定。</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;"> </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,暫時不測定,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">℃以下可保存一個月</font><font face="Calibri">,-70</font><font face="宋體">℃以下可保存三個月。</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>