摘要
目的 探讨单份及16份混合标本2种检测模式对献血者血液病毒核酸检测（nucleic acid test,NAT）效果的影响.方法 2009年2至6月顺序留取北京无偿献血者标本,用诺华Procleix ULTRIO Assay进行单份（ID）或16份混合标本（P16）乙型肝炎病毒（HBV）、丙型肝炎病毒（HCV）和人类免疫缺陷病毒-1（ HIV-1）三项联合核酸检测.单份NAT反应性同时HBsAg、抗-HCV或抗-HIV血清学不合格的标本,血清学合格的单份NAT反应性经双孔NAT复检阳性的标本,以及混合NAT反应性/拆分NAT为阳性的标本,进一步用诺华Procleix HBV、HCV和HIV-1鉴别试剂进行鉴别试验.血清学合格、HBV NAT单独阳性标本进一步用Roche HBV定量实验加以验证和进行病毒含量测定、血清学分析、并进行稀释以模拟是否能被P16-NAT检出.阳性检出率进行四格表连续校正的x2检验.结果 （1）在7613份单份NAT （ID-NAT）标本中,检出NAT阳性26份,ID-NAT阳性率0.34％（26/7613）；（2）在16 064份共1004份P16混合标本NAT（P16-NAT）中,检出NAT阳性27份,P16-NAT阳性率为0.17％ （27/16 064）；（3）在血清学合格标本中,单份检测的NAT单独阳性检出率为0.12％ （9/7438）,高于16份混样检测的NAT单独阳性检出率0.01％ （2/15 750）（x2=11.880,P＜0.05）.9份ID-NAT及2份P16-NAT单独阳性标本经鉴别均为HBV NAT阳性,未检出 HCV NAT单独阳性或HIV NAT单独阳性；（4）9份ID-NAT HBV单独阳性血样模拟P16-NAT,仅有2份可被检出；（5）对8份ID-NAT及2份P16-NAT单独阳性标本进行Roche HBV定量测定,均可确证其核酸检测结果,但病毒含量很低.其中2份HBV病毒含量为472 IU/ml及15 IU/ml,6份含量＜12 IU/ml,另2份原倍不能定量经10倍浓缩处理后测得含量为＜ 12 IU/ml和14.3 IU/ml;（6）11份HBV NAT单独阳性标本中,3份（27.3％）为潜在的窗口期感染,其余8份（72.7％）抗-HBc阳性或抗-HBe阳性,但抗-HBc-IgM均为阴性,为隐匿性感染；（7） P16-NAT初检呈反应性需要进行拆分试验的混合样本比率为2.49％ （25/1004）,其中由血清学合格标本所致初检反应性的混合样本比率为0.20％ （2/1004）.结论 ID-NAT单独阳性检出率高于P16-NAT单独阳性检出率.为避免低病毒含量HBV的漏检,应选用灵敏度高的核酸检测试剂,并尽量采用小标本量混合检测,甚至采用单份检测方式.
：Objective To investigate the effect of individual donation-nucleic acid amplification test （ID-NAT） and minipool of 16 donations-NAT （P16-NAT） on the results of NAT of blood donors.Methods From February 2009 to June 2009,samples randomly collected from voluntary blood donors in Beijing were tested individually or in pooling of 16 donations by the PROCLEIX ULTRIO assay.For ID-NAT reactive samples with HBsAg,anti-HCV,or anti-HIV serologically unqualified,ID-NAT repeat reactive samples with serologically qualified,and P16-NAT reactive and followed resolution ID-NAT reactive samples,were performed for further discriminatory assays for HIV-1,samples and followed resolution ID-NAT reactive samples,were performed for further discriminatory assays for HBV,HCV and HIV-1 discriminatory reagents.Samples which were HBV NAT ＋ alone with serologically qualified were further quantified and confirmed of HBV DNA by Roche HBV quantitative PCR,analyzed by HBV serology and were diluted to simulate if they could be detected in P16-NAT.Results （ 1 ） Among 7613 samples tested by ID-NAT,26 were NAT positive,i.e.the ID-NAT positive rate was 0.34% （ 26/7613 ）. （ 2 ） Among 1004 P16 samples from 16 064 blood donations,27 were NAT positive,i.e.the P16-NAT positive rate was 0.17% （27/16 064）.（3）In serological qualified donations,ID-NAT yield rate （1 in 826,9/7438 ） was much higher than P16-NAT （ 1 in 7875,2/15 750） （x2 =11.880,P 〈 0.05 ）.All these 9 ID-NAT positive and 2 P16-NAT positive donations were discriminated as HBV NAT positive.There were no HCV NAT yield or HIV NAT yield samples. （4） Dilution assay showed only 2 of the 9 （22.22% ） ID-NAT HBV yields were detected by P16-NAT.（5）Eight ID-NAT and 2 P16-NAT positive samples were quantified for HBV DNA and confirmed as HBV NAT yield,although the virus loads were very low：2 samples had HBV viral loads of 15 IU/ml and 472 IU/ml,6 samples 〈 12 IU/ml,and 2 could not be detected in the original samples while had 〈 12 IU/ml and 14.3 IU/ml in the 10 times concentrated samples.（6）Among 11 HBV NAT yield cases,3 （27.3% ） were possible HBV window-period donors with all HBV seromarkers negative,the other 8 （72.7% ） had occult HBV infections with anti-HBc or anti-HBe positive,however anti-HBc IgM negative.（7） The rate of initial P16-NAT reactive pools needed to be further tested by ID-NAT was 2.49%（25/1004）.Initial P16-NAT reactive pools which caused by serologically qualified donations was 0.20%（2/1004）.Conclusions HBV NAT yield cases are detected at a higher frequency with ID-NAT than P16-NAT.In order to avoid samples with low viral loads would be undetected,NAT assay with high sensitivity should be selected and tested in minimized minipool donations or even with individual donation.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2012年第1期53-58,共6页
Chinese Journal of Laboratory Medicine
基金
基金项目：北京市人事局“新世纪百千万人才工程”资助项目（2007-01）
卫生部“卫生公益性科研专项”资助项目（200902008）
