血站核酸检测实施后献血者血液检测模式的研究

Blood Test Patterns for Blood Donors after Nucleic Acid Detection in the Blood Center

目的:探讨血站核酸检测实施后献血者血液检测的模式。方法:对无偿献血者献血后留取的血液样本先进行常规ELISA检测,对31 981阴性样本用罗氏Cobas s201仪器采用6人份混合样本(22 716例)或单人份样本(9265例)的HBV/HCV/HIV三项联合核酸检测。混合检测的具体方法为:采用常规6混合样本核酸检测,与此同时,6混合样本后用聚乙二醇沉淀法进行病毒浓缩再进行核酸检测。单份核酸检测方法为:常规单份核酸检测,在此基础上反应性样本经核酸复检阳性者,进行6倍稀释以模拟6混合样本核酸检测。对混合核酸检测出的核酸阳性样本、混合病毒浓缩核酸法检测出的阳性样本与单份核酸检测经复检核酸阳性样本的阳性率进行统计学分析;对HBV^+者进行补充血清学检测。结果:用6混合样本核酸检测﹙MP-6-NAT﹚法检测22 716份样本,9例HBV^+(0.40‰,9/22 716);同时,用6混合样本病毒浓缩核酸检测法检测同样的样本29例,显示HBV^+﹙1.28‰,29/22716﹚。用单份核酸检测﹙ID-NAT﹚法检测9 265份样本,HBV+样本12例(1.30‰,12/9 265﹚;在此基础上,对这12例HBV^+样本进行6倍稀释以模拟6混合样本核酸检测,只检出4例HBV^+样本。在血清学合格样本中,IDNAT不合格率1.30‰高于MP-6-NAT不合格率0.40‰﹙χ~2=8.11,P<0.05﹚;ID-NAT不合格率1.30‰与6混合样本病毒浓缩NAT不合格率1.28‰比较差异无统计学意义﹙χ~2=0.00,P>0.05﹚。41例ELISA法HBs Ag-HBV DNA+样本,经ELISA法HBsAb、HBcAb确认36例为隐匿性HBV感染者﹙OBI﹚,其中33例HBcAb^+,占OBI的91.66%。5例可能为HBV"窗口期"感染者。未发现HCV RNA、HIV RNA阳性样本。结论:为避免低病毒含量献血者漏检,在献血者血液检测模式为混合样本核酸检测时一定进行混合样本病毒浓缩处理并再进行核酸检测,否则要进行单检的检测模式,以实现更高的病毒核酸检出率;在血清学检测HBsAg基础上,应注重HBcAb血清学检测和加大献血前体检力度:对具有高危行为人群劝导不献血的模式是最简单的模式。

Objective： To investigate the blood test patterns for blood donors after nucleic acid detection in blood center. Methods： The collected blood samples after voluntary blood donors first were detected by conventional ELISA,then 31 981 negative samples were detected via HBV/HCV/HIV combined nucleic acid test of 6 mixed samples（ 22716 cases）or single samples（ 9265 cases） by means of Roche cobas s201 instrument. The combined detection method as follows： the blood samples were assayed by conventional nucleic acid test of 6 mixed samples,at same time,6 mixed samples were treated with polyethylene glycol precipitation method to concentrate the virus,then the nucleic acid test of blood samples was performed; the single detection method as follows： firstly the conventional nucleic acid test of single sample was performed,then the positive reactive samples after re-examination were 6-fold diluted to simulate the nucleic acid test of 6-mixed samples. The positive rate of positive samples detected by combined nucleic acid test,positive samples detected by nucleic acid test of mixed virus concentration and positive samples detected by single nucleic acid test was statistically analyzed. In addition,for HBV＋persons the serological test yet should be performed. Results： In22 716 samples detected by nucleic acid test of 6 mixed samples（ MP-6-NAT）,9 cases were HBV＋（ 0. 40‰,9/22716）;at same time,the detection of same samples by nucleic acid test of mixed sample virus concentration showed 29 cases of HBV＋（ 1. 28‰,29/22716）. In 9265 samples detected by single nucleic acid test（ ID-NAT） 12 cases showed HBV^＋（ 1. 30‰,12/9265）,meanwhile the detection of these 12 samples with HBV＋by 6-fold dilution for virus concentration found only 4 samples with HBV^＋. In serological qualified samples,ID-NAT unqualified rate was 1. 28‰,which was higher than that of MP-6-NAT（ 0. 4‰）（ χ^2= 8. 11,P〈0. 05）; but there was no statistical difference between unqualified rate of ID-NAT and MP-6-NAT（ 1. 3‰ vs 1. 28‰）（ χ^2= 0. 00,P〈0. 05）. In 41 samples with HBsAg-HBV DNA＋detected by ELISA,36 samples were confirmed to be occult HBV infective（ OBI） by HBsAb,HBcAb test of ELISA; out of these 41 samples,33 samples showed HBcAb^＋（ 91. 66% of OBI）,5 might be HBV ＂ window period＂ infective,moreover the HCV RNA and HIV RNA positive samples were not found. Conclusion： To avoid the missdiagnosis of donors with low level of virus,the nucleic acid test must be carried out after virus concentration of mixed samples when the blood test pattern of donors is nucleic acid test of mixed samples,otherwise the single nucleic acid test must be performed to obtain more high detected rate of virus nucleic acid. The HBcAb serologic test and physical examination of donors before blood donation must be enhanced on basis of serological test of HBsAg; for high risk people,the persuading no blood donation is simplest pattern.