用于HCV RNA逆转录聚合酶链反应测定的血清(浆)样本的质量控制

Quality contral of plasma(serum) specimen for the detection of HCV RNA with PCR

目的确定用于丙型肝炎病毒（ＨＣＶ）ＲＮＡ测定血清（浆）样本的最佳收集贮存条件。方法采集ＨＣＶＲＮＡ阳性的血样按设定的不同条件处理，用荧光定量ＰＣＲ试剂测定ＨＣＶＲＮＡ。结累血凝集后２小时内离心分高血清ＨＣＶＲＮＡ含量无明显改变（降低１０.４２％）；４小时变化明显（降低４０．４９％）。采用抗凝剂的血浆管ＨＣＶＲＮＡ含量显著高于血清管（拘檬酸钠抗凝管高出４０．９７％，ＥＤＴＡ抗凝管高出５３．１４％）。反复冻融无明显变化；样本在-２０Ｃ贮存１年，ＨＣＶＲＮＡ降低１５６％。结论结果表明制备和贮存用于ＨＣＶＲＮＡ测定临床样本条件是很重要的。应尽量用ＥＤＴＡ抗凝管，３小时内分高血浆；如不用抗凝剂，２小时内离心分离血清；样本长期贮存应在－７０℃。

Objective To define the optimal collection and storage parameters of serum (plasma) forHCV RNA detection. Methods The serum(plasma) obtained from HCV RNA positive patients by differentmeans and different processing and storage conditions, then HCV RNA was detected with the flurescentquantitative PCR. Results The centrifugstion was performed witnin 2 hours aft6r formation of the clot,loss of HCV RNA titers was 10.42% and significant loss of HCV RNA tit6rs 40.49% was observed after4h. The HCV RNA tit6rs in plasma samples with anticoagulants were much higher than that in serumsamples(citrate sodium-anticoagulized 40.97%, EDTA-anticoagLilated 53.14%). There was no loss of HCV RNAwith op to three fi'eeZe-thaw cycles. A significant decrease in HCV RNA(15.6%) was oborved at -20C afterone y6ar. Conclusions The results suggested that it is important to process and store clinical samples forHCV RNA detection. The EDTA-K2 as anticoagUlants the plasma must be collected within 3h, and theserum must be prepared within 2 hours after hemosposia. It is essential to store the sample under -70Cfor the stability of HCV RNA in a longer time.