Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, USA.
J Clin Microbiol. 2013 Jan;51(1):95-100. doi: 10.1128/JCM.02155-12. Epub 2012 Oct 24.
The Abbott HBV RUO Sequencing assay (Abbott Molecular Inc., Des Plaines, IL), which combines automated sample processing, real-time PCR, and bidirectional DNA sequencing, was evaluated for detection of nucleos(t)ide analogue (NA) resistance-associated mutations located in the hepatitis B virus (HBV) polymerase (Pol) gene. Interpretive software from the assay manufacturer was modified to allow interrogation of the overlapping HBV surface (S) gene sequence for HBV genotype determination and detection of immune escape mutations. Analytical sensitivity (detection and sequencing) of the assay was determined to be 103.9 IU/ml (95% confidence interval [CI], 80.0 to 173.3) for HBV genotype A. Testing of commercially available HBV genotype panels consisting of 23 individual members yielded complete agreement between expected results and results obtained from the laboratory-developed HBV genotype library. Excellent specificity was observed among clinical specimens with serologic or molecular markers for various unrelated blood-borne viruses (n = 6) and sera obtained from healthy, HBV-negative blood donors (n = 20). Retrospectively selected clinical specimens tested by a commercial reference laboratory HBV sequencing assay (n = 54) or the Trugene HBV Genotyping kit (n = 7) and the Abbott HBV RUO Sequencing assay showed minor differences in detection and reporting of NA resistance-associated mutations in 7 of 61 (11.5%) specimens but complete agreement of genotype results. The Abbott HBV RUO Sequencing assay provided a convenient and efficient assay workflow suitable for routine clinical laboratory use, with the flexibility to be modified for customized detection of NA resistance-associated mutations, HBV genotype determination, and detection of immune escape mutations from a single contiguous HBV sequence.
Abbott HBV RUO 测序检测（雅培分子公司，德斯普兰斯，IL），结合自动化样本处理、实时 PCR 和双向 DNA 测序，用于检测乙型肝炎病毒（HBV）聚合酶（Pol）基因中核苷（酸）类似物（NA）耐药相关突变。该检测的制造商解释软件进行了修改，允许对重叠的 HBV 表面（S）基因序列进行查询，以确定 HBV 基因型并检测免疫逃逸突变。该检测的分析灵敏度（检测和测序）对于 HBV 基因型 A 为 103.9 IU/ml（95%置信区间 [CI]，80.0 至 173.3）。对由 23 个个体成员组成的商业可用 HBV 基因型面板进行测试，结果与实验室开发的 HBV 基因型文库获得的结果完全一致。在具有各种无关血液传播病毒的血清学或分子标志物的临床标本（n=6）和来自健康、HBV 阴性献血者的血清（n=20）中观察到极好的特异性。通过商业参考实验室 HBV 测序检测（n=54）或 TruGene HBV 基因分型试剂盒（n=7）和 Abbott HBV RUO 测序检测进行的回顾性选择的临床标本在 61 个标本中的 7 个（11.5%）标本中检测和报告 NA 耐药相关突变方面存在微小差异，但基因型结果完全一致。Abbott HBV RUO 测序检测提供了一种方便且高效的检测工作流程，适用于常规临床实验室使用，并且具有灵活性，可以进行修改，以从单个连续 HBV 序列中定制检测 NA 耐药相关突变、HBV 基因型确定和检测免疫逃逸突变。
