ARUP Institute for Clinical and Experimental Pathology, ARUP Laboratories, Salt Lake City, UT 84108, USA.
J Virol Methods. 2011 Oct;177(1):31-7. doi: 10.1016/j.jviromet.2011.06.009. Epub 2011 Jun 23.
The objective of this study was to develop a DNA sequencing assay that examines sensitively and reliably all conserved domains of the reverse transcriptase-encoding region of the HBV genome for antiviral resistance-associated mutations while simultaneously producing ample information for precise genotyping and determination of HBsAg mutation. This assay was used to examine 1000 de-identified HBV DNA positive samples with known viral loads from a broad-based, unselected patient population from across the United States. Of these, 946 were assayed successfully. Antiviral resistance-associated mutations were identified in 104 samples. The escape mutation sG145R in the surface antigen was identified in 0.8% of patient samples. Infections with genotypes A, B, C, D, E, F, G and H were observed in 36.6%, 19.6%, 21.7%, 13.5%, 3.6%, 0.7%, 2.2%, and 0.5% of patient samples respectively. Fifteen samples (1.6%) appeared to harbor infections with multiple genotypes as shown by the presence of double peaks throughout sequence electropherograms. The limit of detection of this assay was approximately 150IU/mL.
本研究的目的是开发一种 DNA 测序检测方法，该方法能够灵敏且可靠地检测乙型肝炎病毒（HBV）基因组中逆转录酶编码区域的所有保守结构域，以发现与抗病毒药物耐药相关的突变，同时为准确的基因分型和 HBsAg 突变的确定提供充分的信息。该检测方法用于检测来自美国各地广泛、未选择的患者人群中具有已知病毒载量的 1000 份经鉴定的 HBV DNA 阳性样本。其中，946 份样本成功进行了检测。在 104 份样本中发现了与抗病毒药物耐药相关的突变。在 0.8%的患者样本中发现了表面抗原中的逃逸突变 sG145R。在患者样本中分别观察到基因型 A、B、C、D、E、F、G 和 H 的感染率为 36.6%、19.6%、21.7%、13.5%、3.6%、0.7%、2.2%和 0.5%。15 份（1.6%）样本似乎存在多重基因型感染，这可以从整个序列电泳图谱中存在双峰看出。该检测方法的检测下限约为 150IU/mL。
