逆转录聚合酶链反应检测HCV RNA血标本保存条件分析

Analysis of reserved conditions of blood sample for detection of HCV RNA with RT-PCR

目的 确定用于丙型肝炎病毒 (HCV)RNA逆转录聚合酶链反应测定的血清 (浆 )标本的最佳收集和保存条件。方法 按设定的不同条件处理 ,用逆转录聚合酶链反应测定HCVRNA。结果 用高压消毒的离心管与未用高压消毒的离心管收集、保存HCVRNA血清 ,其阳性检出率有差异 (P <0 0 5 )。血凝固后 4h内离心分离血清对HCVRNA阳性检出率不造成明显改变 (阳性率为 85 % ) ;12h后检测则变化明显 (阳性率降至 75 % )。采用枸橼酸钠和EDTA K2抗凝剂的血浆管HCVRNA检出率分别为 90 %和 95 % ,显著高于血清管 (为 6 5 % )。反复冻融影响标本阳性检出率 ,标本在 - 2 0℃保存 3个月或半年几乎不影响检测结果。结论 收集和保存HCVRNA检测标本应尽量用EDTA抗凝管 ,在 4h内分离血浆 ;若用血清标本 ,最好在 4h之内分离血清 ,标本长期保存应分装且存放在 - 70℃。

Objective To investigate the optimal collection and storage parameters of plasma (serum) for HCV RNA detection. Methods The serum (plasma) obtained from HCV RNA positive by different means and different processing and storage conditions, then HCV RNA was detected with RT-PCR. Results When HCV RNA serum is collected and reserved with disinfectant tube and infectant tube. The centrifugation was performed within 4 hours after formation of the clot, the positive rate of HCV RNA was 85%; the significart change of HCV RNA positive rate 75% was observed after 12 h. The positive rate of HCV RNA in plasma with anticoagulants were much higher than that in serum samples (citrate sodium anticoagulized 90%, EDTA-anticoagulated 95%). A significant decrease in HCV RNA positive rate was observed with up to three freeze thaw cycles. There was no change of HCV RNA at -20℃ after three months or six months. Conclusion It is important to process and store clinical samples for HCV RNA detection. The EDTA-K 2 as anticoagulants the plasma must be collected within 24 h; and the serum must be prepared within 4 h after hemosposia. It is essential to store the sample under -70℃ for the stability of HCV RNA for a longer time.