干血斑用于HIV抗体筛查(免疫层析法)的评价
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摘要
目的研究建立干血斑(DBS)样本用于艾滋病病毒(HIV)抗体筛查(免疫层析法)的方法,为扩大HIV检测提供一个简便可行的筛查方法。方法制备系列稀释20～210干血斑与配对血浆样本,对DBS进行洗脱后,研究来自两家不同生产商的人类免疫缺陷病毒(HIV1/2)抗体检测试剂盒(胶体金法)的检测限,并以最低检出限为依据优化选择用于该两种试剂的最适DBS洗脱液体积和加样体积,通过310份已知样本与1 304份临床样本对该方法进行验证与应用。结果来自两个不同生产商的试剂检测限一致,但同一试剂下配对血浆样本与DBS样本检测限不同(血浆为20～27,DBS为20～23);两种试剂优化选择出的最适DBS洗脱液体积(200μL)和加样体积(150μL)一致,可将最低检测限降低至24;两种试剂进行方法验证的结果一致,阳性一致率均为95.45%(105/110),阴性一致率均为100%(200/200),总一致率均为98.39%(305/310),Kappa值0.964(P<0.001);两种试剂进行临床应用结果完全一致,来自北京市男男性行为者(MSM)1 304份临床DBS样本中(以随访确认结果为金标准),免疫层析法的敏感性为92.31%(120/130),特异性为100%(1 174/1 174),检测准确率为99.23%(1 294/1 304);与酶联免疫吸附试验检测结果相比,免疫层析法存在7.69%(10/130)弱阳性漏检,无假阳性出现。结论DBS样本的HIV快速筛查方法简便可行,无假阳性,但存在弱阳性漏检,其应用存在地区与人群差异,更适用于落后偏远、弱阳性率低的地区与人群筛查。
Objective To establish a simple and feasible HIV antibody screening method(immune-chromatography)based on DBS(dry blood spot samples)so as to expand HIV testing in China.Methods A series of diluted 20-210 dry blood spots was prepared with matching plasma samples,and DBS samples were eluted to study the detection limit of human immunodeficiency virus(HIV1/2)antibody detection kit(colloidal gold method)from two different manufacturers,and to optimize DBS elution volume and the test sample volume according to the minimum detection limit,in order to establish rapid screening method for HIV antibodies of DBS.310 samples of known infection status and 1304 samples from clinical applications were used to verify and apply the method.Results The detection limit of kits from two different manufacturers was the same,but the detection limit of the matched plasma samples and DBS samples was different based on the same kits(plasma was 20-27 and DBS was 20-23).The optimal DBS elution volume(200μL)and the test sample volume(150μL)of the two kits were the same.The results of the method verification of the two reagents were consistent,including 95.45%(105/110)of the positive coincidence rate,100%(200/200)of the negative coincidence rate,and 98.39%(305/310)of the total coincidence rate,and 0.964(P<0.001)of the Kappa value.The clinical application results of the two reagents were completely consistent.Among 1304 clinical DBS samples from the MSM people in Beijing,the sensitivity was 92.31%(120/130),the specificity was 100%(1 174/1 174),and the detection accuracy was 99.23%(1294/1304).Compared with the ELISA test results of DBS,the weak positiveleakage test rate was 7.69%(10/130).Conclusion Rapid HIV screening method used to test DBS samples is simple,feasible,and no false positives.However,it has a weak positive leak detection,and there are regional and population differences in its application.It is more suitable for HIV screening in the remote areas and the population with a low rate of weak positive.
Objective To establish a simple and feasible HIV antibody screening method(immune-chromatography)based on DBS(dry blood spot samples)so as to expand HIV testing in China.Methods A series of diluted 20-210 dry blood spots was prepared with matching plasma samples,and DBS samples were eluted to study the detection limit of human immunodeficiency virus(HIV1/2)antibody detection kit(colloidal gold method)from two different manufacturers,and to optimize DBS elution volume and the test sample volume according to the minimum detection limit,in order to establish rapid screening method for HIV antibodies of DBS.310 samples of known infection status and 1304 samples from clinical applications were used to verify and apply the method.Results The detection limit of kits from two different manufacturers was the same,but the detection limit of the matched plasma samples and DBS samples was different based on the same kits(plasma was 20-27 and DBS was 20-23).The optimal DBS elution volume(200μL)and the test sample volume(150μL)of the two kits were the same.The results of the method verification of the two reagents were consistent,including 95.45%(105/110)of the positive coincidence rate,100%(200/200)of the negative coincidence rate,and 98.39%(305/310)of the total coincidence rate,and 0.964(P<0.001)of the Kappa value.The clinical application results of the two reagents were completely consistent.Among 1304 clinical DBS samples from the MSM people in Beijing,the sensitivity was 92.31%(120/130),the specificity was 100%(1 174/1 174),and the detection accuracy was 99.23%(1294/1304).Compared with the ELISA test results of DBS,the weak positiveleakage test rate was 7.69%(10/130).Conclusion Rapid HIV screening method used to test DBS samples is simple,feasible,and no false positives.However,it has a weak positive leak detection,and there are regional and population differences in its application.It is more suitable for HIV screening in the remote areas and the population with a low rate of weak positive.
引文
[1]赫晓霞,蒋岩.干血斑用于HIV检测的研究与展望[J].中国艾滋病性病,2018,24(11):1171-1173,1177.
[2]He X,Liu G,Xia D,et al.An innovative HIV testing service using the internet:Anonymous urine delivery testing service at drugstores in Beijing,China[J].PLoS One,2018,13(2):e0192255.
[3]He XX,Feng X,Liu P,et al.An Innovative Vending MachineBased HIV Testing and Intervention Service in China:Anonymous Urine Collection Kits Distributed at Universities[Z].JIAS,Under review,2018.
[4]Zou H,Wu Z,Yu J,et al.Internet-Facilitated,Voluntary Counseling and Testing(VCT)Clinic-Based HIV Testing among Men Who Have Sex with Men in China[J].PLoS One,2013(8):e51919.
[5]Zhang GL,Zhang XY,Liang X.Results analysis of anonymous HIV antibody urine delivering detection based on the pharmacy.Chin[J].J AIDS STD,2016(10):810-812.
[6]赫晓霞,粟斌,程焕义,等.干血斑样本用于HIV抗体检测方法学优化与应用[J].中国艾滋病性病,2018,24(6):431-435.
[7]中国疾病预防控制中心.全国艾滋病检测技术规范[S].2015.
[8]郭颜艳.ELISA、PA、ICA检测人类免疫缺陷病毒抗体的结果比较[J].世界临床医学,2017,1(11):239.
[9]Pinkerton SD.How many sexually-acquired HIV infection in the USA aredue to acute-phase HIV transmission[J].AIDS,2007,21(12):1625.
[10]宋炜路,于茂河,李志军,等.男男性行为者HIV感染急性期筛查诊断和治疗工作模式探索[J].中国艾滋病性病,2017,23(12):1124-1127.
[11]李秀华,宋爱京,王佑春.HIV抗体快速检测试剂的临床评价[J].中华实验与临床病毒学杂志,2006,9(3):279-281.
[12]可秋萍,娄莹,刘传振,等.危重症手足口病患儿的早期预警指标[J].实用儿科医学杂志,2011,26(10):804-805.
[13]王希英,张国林.免疫层析试验检测抗-HIV假阳性原因分析[J].中国误诊学杂志,2007,1(7):246-247.
[14]夏冬艳,刘国武,曾吉,等.北京市HIV抗体快速检测为阳性的男男性行为人群后续确认检测行为的相关因素分析[J].中华预防医学杂志,2016,2(2):153-157.
[15]施玉华,苏莹珍,方清艳,等.HIV抗体快速检测在自愿咨询检测中的应用[J].传染病信息,2014,2(1):29-34.
[16]闫莉,肖培培,闫红静.HIV自我检测技术研究进展[J].中华预防医学杂志,2017,11(51):1053-1056.
[2]He X,Liu G,Xia D,et al.An innovative HIV testing service using the internet:Anonymous urine delivery testing service at drugstores in Beijing,China[J].PLoS One,2018,13(2):e0192255.
[3]He XX,Feng X,Liu P,et al.An Innovative Vending MachineBased HIV Testing and Intervention Service in China:Anonymous Urine Collection Kits Distributed at Universities[Z].JIAS,Under review,2018.
[4]Zou H,Wu Z,Yu J,et al.Internet-Facilitated,Voluntary Counseling and Testing(VCT)Clinic-Based HIV Testing among Men Who Have Sex with Men in China[J].PLoS One,2013(8):e51919.
[5]Zhang GL,Zhang XY,Liang X.Results analysis of anonymous HIV antibody urine delivering detection based on the pharmacy.Chin[J].J AIDS STD,2016(10):810-812.
[6]赫晓霞,粟斌,程焕义,等.干血斑样本用于HIV抗体检测方法学优化与应用[J].中国艾滋病性病,2018,24(6):431-435.
[7]中国疾病预防控制中心.全国艾滋病检测技术规范[S].2015.
[8]郭颜艳.ELISA、PA、ICA检测人类免疫缺陷病毒抗体的结果比较[J].世界临床医学,2017,1(11):239.
[9]Pinkerton SD.How many sexually-acquired HIV infection in the USA aredue to acute-phase HIV transmission[J].AIDS,2007,21(12):1625.
[10]宋炜路,于茂河,李志军,等.男男性行为者HIV感染急性期筛查诊断和治疗工作模式探索[J].中国艾滋病性病,2017,23(12):1124-1127.
[11]李秀华,宋爱京,王佑春.HIV抗体快速检测试剂的临床评价[J].中华实验与临床病毒学杂志,2006,9(3):279-281.
[12]可秋萍,娄莹,刘传振,等.危重症手足口病患儿的早期预警指标[J].实用儿科医学杂志,2011,26(10):804-805.
[13]王希英,张国林.免疫层析试验检测抗-HIV假阳性原因分析[J].中国误诊学杂志,2007,1(7):246-247.
[14]夏冬艳,刘国武,曾吉,等.北京市HIV抗体快速检测为阳性的男男性行为人群后续确认检测行为的相关因素分析[J].中华预防医学杂志,2016,2(2):153-157.
[15]施玉华,苏莹珍,方清艳,等.HIV抗体快速检测在自愿咨询检测中的应用[J].传染病信息,2014,2(1):29-34.
[16]闫莉,肖培培,闫红静.HIV自我检测技术研究进展[J].中华预防医学杂志,2017,11(51):1053-1056.