Garrido Carolina, Zahonero Natalia, Corral Angélica, Arredondo Miguel, Soriano Vincent, de Mendoza Carmen
Department of Infectious Diseases, Hospital Carlos III, Calle Sinesio Delgado 10, Madrid 28029, Spain.
J Clin Microbiol. 2009 Apr;47(4):1031-6. doi: 10.1128/JCM.02099-08. Epub 2009 Feb 4.
The plasma human immunodeficiency virus (HIV) RNA load is used in the clinical routine for the monitoring of HIV infection and the patient's response to antiretroviral therapy. Other body fluids or dried blood spots (DBS) can be used, however, to assess the level of viremia. The use of DBS may be especially helpful for the monitoring of HIV-infected patients in resource-poor settings, where access to adequate laboratory facilities is often difficult. However, the correlation between the HIV RNA levels in plasma and those in DBSs has not been well established. Paired plasma and DBS samples obtained from HIV type 1 (HIV-1)-infected patients were tested for HIV RNA copy numbers by using two different commercial assays, the Nuclisens EasyQ HIV-1 (version 1.1) test (the Nuclisens test; Biomerieux) and the m2000rt RealTime HIV test (the m2000rt test; Abbott). Nucleic acid extraction was performed manually by using either the Nuclisens isolation kit (which uses the Boom methodology) or the m2000rt sample preparation kit (an iron particle-based method). A total of 103 paired plasma and DBS samples were tested. Viral load results were obtained for 97 (94.2%) samples with the Nuclisens isolation kit and 81 (78.6%) samples with the m2000rt kit. The overall correlation between the RNA loads in plasma and DBS was good, although better results were obtained by the Nuclisens test (R(2) = 0.87, P < 0.001) than by the m2000rt test (R(2) = 0.70, P < 0.001). While the specificities were excellent and similar for both the Nuclisens and the m2000rt tests (97.1% and 100%, respectively), the sensitivity was greater by the Nuclisens test than by the m2000rt test (75.8% and 56.6%, respectively). Overall, the viral loads in DBS tended to be lower than those in plasma, with mean differences of 0.3 log unit (standard deviation, 0.5 log unit) and 0.76 log unit (standard deviation, 0.8 log unit) for the Nuclisens and the m2000rt tests, respectively. The levels of agreement between the measurements in plasma and DBS were assessed by using the Bland-Altman plot for each assay. The Nuclisens test gave results within its defined limits (-0.65 to 1.26) for 95.9% of the samples, while the m2000rt test gave results within its limits (-0.83 to 2.33) for 100% of the samples. In summary, the HIV-1 load can accurately be quantified by testing DBS by either the Nuclisens or the m2000rt test, although the Nuclisens test may outperform the m2000rt test when nucleic acids are extracted manually.
血浆人类免疫缺陷病毒(HIV)RNA载量在临床常规中用于监测HIV感染以及患者对抗逆转录病毒疗法的反应。不过,也可以使用其他体液或干血斑(DBS)来评估病毒血症水平。对于资源匮乏地区的HIV感染患者监测而言,使用DBS可能特别有用,因为在这些地区往往难以获得足够的实验室设施。然而,血浆和DBS中HIV RNA水平之间的相关性尚未完全确立。我们使用两种不同的商业检测方法,即Nuclisens EasyQ HIV-1(版本1.1)检测(Nuclisens检测;生物梅里埃公司)和m2000rt实时HIV检测(m2000rt检测;雅培公司),对从1型HIV(HIV-1)感染患者获得的配对血浆和DBS样本进行HIV RNA拷贝数检测。使用Nuclisens分离试剂盒(采用Boom方法)或m2000rt样本制备试剂盒(基于铁颗粒的方法)手动进行核酸提取。总共检测了103对血浆和DBS样本。使用Nuclisens分离试剂盒对97个(94.2%)样本获得了病毒载量结果,使用m2000rt试剂盒对81个(78.6%)样本获得了病毒载量结果。血浆和DBS中RNA载量之间的总体相关性良好,不过Nuclisens检测(R² = 0.87,P < 0.001)获得的结果比m2000rt检测(R² = 0.70,P < 0.001)更好。虽然Nuclisens和m2000rt检测的特异性都非常好且相似(分别为97.1%和100%),但Nuclisens检测的灵敏度高于m2000rt检测(分别为75.8%和56.6%)。总体而言,DBS中的病毒载量往往低于血浆中的病毒载量,Nuclisens检测和m2000rt检测的平均差异分别为0.3 log单位(标准差,0.5 log单位)和0.76 log单位(标准差,0.8 log单位)。通过使用每种检测方法的Bland-Altman图评估血浆和DBS测量值之间的一致性水平。Nuclisens检测对95.9%的样本给出的结果在其定义的限值(-0.65至1.26)内,而m2000rt检测对100%的样本给出的结果在其限值(-0.83至2.33)内。总之,通过Nuclisens或m2000rt检测对DBS进行检测可以准确量化HIV-1载量,不过在手动提取核酸时,Nuclisens检测可能优于m2000rt检测。