LAg-Avidity EIA用于中国HIV-1新发感染率估算的适用性研究

发布时间：2018-07-10 19:44

本文选题：人类免疫缺陷病毒Ⅰ型 + 限制性抗原亲和力酶联免疫方法　；参考：《中国疾病预防控制中心》2013年博士论文

【摘要】：研究背景利用实验室检测技术从横断而样本中发现HIV-1新近感染样本,从而估算HIV-1新发感染率的实验室方法越来越受到重视。目前,普遍使用的BED方法存在影响因素多,假近期感染率较高等问题；为了解决这些问题,最近国际上研发出了一种新的亲和力方法——限制性抗原亲和力酶联免疫方法(Limiting antigen avidity enzyme immunoassay, LAg-Avidity EIA),经初步评价该方法受影响因素较少,误判率大大降低,可以明显提高估算的HIV-1新发感染率的准确性。研究目的一、评价LAg-Avidity试剂盒的重复性和准确度,探讨其在中国推广应用的可行性；二、获得LAg-Avidity方法应用于中国HIV-1新发感染率计算所必需的窗口期参数；三、研究LAg-Avidity方法应用于中国HIV-1新发感染率计算所必需的假近期感染率参数,初步探讨CD4+T淋巴细胞计数对LAg-Avidity方法的影响；四、研究抗病毒治疗对LAg-Avidity方法准确性的影响。研究方法一、采用试剂盒自带的质控品和校准品通过检测精密度和孔问精密度指标,评价LAg-Avidity试剂的重复性,通过四位实验员之间刘BOCA101盘检测结果的一致性,四位实验员对该标准盘检测结果的平均值与该标准盘期望值结果的一致性,进一步评价试剂盒的重复性；同时,BOCA101盘的判读结果用于评价LAg-Avidity试剂的准确度。二、通过检测531份血清学阳转队列样本(151个血清学阳转感染者),利用SPSS17.0软件对检测结果进行统计学分析,研究LAg-Avidity方法的窗口期。三、通过检测513份已知长期感染(诊断时间大于方法学两倍窗口期)并且未进行抗病毒治疗的感染者样本,研究LAg-Avidity方法的假近期感染率参数。四、通过对720份抗病毒治疗不同时间的病人样本的检测,计算误判比例,研究抗病毒治疗时间以及治疗后病毒载量值与LAg-Avidity方法误判的关系。研究结果一、检测精密度结果显示,经过校准品校准以后,试剂盒低阳对照和高阳对照的ODn值的CV小于10%；不同实验员实验结果的一致性较好,四个实验员实验结果的平均值与美国疾病预防控制中心提供的实验结果的相关性较高,R2为0.9849；孔间精密度结果显示,大批量生产的试剂盒可能存在酶联板包被量不均匀的情况。准确性检验中,试剂盒对BOCA101盘评判结果与预期值完全一致。二、LAg-Avidity方法的假近期感染率参数为0.39%(2/513,95%CI0.00%-0.93%),不受CD4+T细胞计数影响(0/52)。三、当方法学的临界值取1.0时,LAg-Avidity方法的窗口期参数为126天。四、抗病毒治疗对LAg-Avidity方法影响较大,误判率为10.00%(72/720,95%CI7.81%-12.19%),误判率和治疗时间没有相关性,治疗以后病毒载量值越低(小于等于1000copies/ml)越容易造成误判。研究结论一、大规模生产的LAg-Avidity试剂盒重复性较好,试剂盒准确度较高,可以在中国推广应用；但试剂盒质量仍有提高的空间。二、初步获得了LAg-Avidity方法用于中国HIV-1新发感染率估算的核心参数,窗口期为126天,假近期感染率为0.39%；与BED方法相比假近期感染率更低,可以使估算的新发感染率更接近真实水平。三、LAg-Avidity方法不受CD4+T淋巴细胞计数的影响,但易受抗病毒治疗的影响,在横断面调查时,应删除已经进行抗病毒治疗的病人样本。
[Abstract]:Background of the study

HIV - 1 newly infected samples were found in the samples from cross - cutting using laboratory testing techniques , so that the method of estimating HIV - 1 infection rate was more and more attention . At present , there are many problems affecting the commonly used BED method , such as many factors , higher false - term infection rate and higher infection rate .
In order to solve these problems , a new affinity method _ restriction antigen avidity enzyme immunoassay ( LAg - Avidin EIA ) has been developed recently .

Purpose of study

1 . To evaluate the repeatability and accuracy of LAg - Avifauna kit and to explore the feasibility of its popularization and application in China ;

secondly , obtaining a LAg - Avifauna method which is applied to the window period parameters necessary for calculating the new infection rate of HIV - 1 in China ;

Thirdly , the study of LAg - Avifauna method was applied to the calculation of HIV - 1 infection rate in China , and the effect of CD4 + T lymphocyte count on the LAg - Avifauna method was discussed .

Fourthly , the effect of antiviral therapy on the accuracy of LAg - Avidin method was studied .

Research Methods

1 , the repeatability of the LAg - Avidin reagent is evaluated by detecting the precision and the hole - to - precision index by using the quality control substance and the calibrator provided by the kit , and the consistency of the average value of the detection result of the standard disc and the expected value of the standard disc is evaluated by four experimenter , and the repeatability of the kit is further evaluated ;
At the same time , the interpretation of the BOCA101 disc was used to evaluate the accuracy of the LAg - Avidin reagent .

Secondly , through the detection of 531 serologic positive transfer cohort samples ( 151 serologic positive patients ) , the detection results were analyzed statistically by SPSS 17.0 software , and the window period of LAg - Avidin method was studied .

3 . The false - recent infection rate parameters of the LAg - Avidin method were investigated by detecting 513 known long - term infections ( diagnosis time greater than twice the window period of the methodology ) and did not perform anti - viral treatment .

Fourthly , the relationship between the anti - virus treatment time and the value of post - treatment virus load and the LAg - Avidin method was studied by detecting the sample of 720 patients with different time .

Results of the study

1 , the detection precision result shows that after the calibration of the calibration sample , the CV of the ODn value of the kit low - positive control and the high - yang control is less than 10 % ;
The results showed that the correlation between the results of the four experimenters was higher than that of the American Centers for Disease Control and Prevention , and R2 was 0.998 ;
The results of inter - hole precision showed that the kit of mass production may be non - uniform in enzyme linked plates . In the accuracy test , the results of the kit ' s evaluation of BOCA101 disc are completely consistent with the expected value .

The infection rate was 0.39 % ( 2 / 513 , 95 % CI 0.00 % - 0.93 % ) in the LAg - Avidin method , which was not affected by CD4 + T cell count ( 0 / 52 ) .

Thirdly , when the critical value of methodology is 1.0 , the window period parameter of the LAg - Avidin method is 126 days .

4 . The effect of antiviral therapy on the LAg - Avifauna method was greater , the rate of misjudgment was 10.00 % ( 72 / 720 , 95 % CI 7.81 % - 12.19 % ) , the rate of misjudgment and treatment time were not correlated , the lower the value of viral load after treatment ( less than or equal to 1000 copies / ml ) , the easier to misjudgment .

Conclusions of the study

firstly , the repeatability of the LAg - Avifauna kit produced by mass production is good , the accuracy of the kit is high , and the LAg - Avifauna kit can be popularized and applied in China ;
but the quality of the kit still has improved space .

Secondly , the core parameters of the LAg - Avifauna method for HIV - 1 infection in China were obtained . The window duration was 126 days , and the infection rate was 0.39 % .
涓嶣ED鏂规硶鐩告瘮鍋囪繎鏈熸劅鏌撶巼鏇翠綆,鍙互浣夸及绠楃殑鏂板彂鎰熸煋鐜囨洿鎺ヨ繎鐪熷疄姘村钩.

3 . The LAg - Avifauna method is not affected by CD4 + T lymphocyte count , but is susceptible to antiviral therapy . In cross - sectional investigation , patients with antiviral therapy should be deleted .
【学位授予单位】：中国疾病预防控制中心
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R512.91

【参考文献】