Unidad de Investigacion Medica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatria, Centro Medico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Delegacion Cuauhtemoc, Mexico City 06720, Mexico.
Arch Virol. 2010 Jul;155(7):1117-25. doi: 10.1007/s00705-010-0696-y. Epub 2010 May 23.
In third-world countries, dried blood samples (DBS) are a convenient alternative to plasma for monitoring viral load during HIV-1 therapy. In this study, we evaluated the feasibility of using DBS to perform HIV-1 drug resistance genotyping in a ViroSeq assay in which the protease and reverse transcriptase regions of the pol gene are analyzed. Fifty-seven antiretroviral genotypes from plasma samples were tested, and drug resistance genotypes were determined. Only 38.6% paired DBS samples were sequenced. Failure to amplify DNA from DBS samples generally correlated with plasma viral loads below log(10) 5.1. The majority of the mutations identified in plasma pol sequences were also found in their DBS counterpart, with a concordance in genotype interpretation of 96.4%. Several factors were identified that could potentially improve both the sensitivity and the quality of genotype data, such as sample storage conditions and sequence analysis. Therefore, DBS sampling is useful to determine viral load and drug resistance genotypes in HIV patients.
在第三世界国家，干血斑（DBS）是监测 HIV-1 治疗期间病毒载量的一种替代血浆的便利方法。在这项研究中，我们评估了使用 DBS 进行 HIV-1 耐药性基因分型的可行性，其中分析了 pol 基因的蛋白酶和逆转录酶区域。对 57 份来自血浆样本的抗逆转录病毒基因型进行了检测，并确定了耐药基因型。只有 38.6%的配对 DBS 样本进行了测序。从 DBS 样本中扩增 DNA 的失败通常与血浆病毒载量低于 log(10)5.1 相关。在血浆 pol 序列中发现的大多数突变也存在于它们的 DBS 对应物中，基因型解释的一致性为 96.4%。确定了一些潜在因素，可以提高基因型数据的灵敏度和质量，例如样本储存条件和序列分析。因此，DBS 采样可用于确定 HIV 患者的病毒载量和耐药基因型。
