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首页> 外文OA文献 >Application of a Novel, Rapid, and Sensitive Oligonucleotide Ligation Assay for Detection of Cancer-Predicting Mutations in the Precore and Basal Core Promoter of Hepatitis B Virus▿
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Application of a Novel, Rapid, and Sensitive Oligonucleotide Ligation Assay for Detection of Cancer-Predicting Mutations in the Precore and Basal Core Promoter of Hepatitis B Virus▿

机译：一种新颖，快速，灵敏的寡核苷酸连接法在检测乙型肝炎病毒前核心和基础核心启动子中的癌症预测突变中的应用▿

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Hepatocellular carcinoma (HCC) and cirrhosis are important causes of mortality worldwide. Persistent hepatitis B virus (HBV) infection is a major cause of these diseases. Double mutations in the basal core promoter (BCP) (A1762T and G1764A) and precore (pre-C) (G1896A) regions of the virus are associated with progression to HCC. The current study is aimed at developing a simple method for screening and detecting BCP and pre-C mutations in HBV carriers. We have developed and validated an oligonucleotide ligation assay (OLA) to detect point mutations in the HBV core gene. We have applied OLA methods to samples from HBV-infected carriers recruited from the Gambia Liver Cancer Study (GLCS) comprising asymptomatic HBsAg carriers, patients with cirrhosis, and patients with HCC. We observed an 89.3% and 95.8% concordance between the OLA and DNA sequencing for BCP and pre-C mutations, respectively. OLA detected the mutations in single-strain infections and in infections with mixtures of wild-type and mutant viruses under conditions where sequencing detected only the single dominant strains. BCP mutations were detected in 75.7% of patients with advanced liver disease (cirrhosis/HCC) compared to 47.6% of asymptomatic carriers, while pre-C mutations were detected in 34.5% of advanced liver disease patients and in 47.6% of asymptomatic HBsAg carriers. There was a significant association between the presence of BCP mutations and advanced liver disease. In conclusion, OLA is a simple, economical, and reliable assay for detection of pre-C and BCP mutations. Its application can lead to improvement in diagnosis and clinical care in regions where HBV is endemic.
机译：肝细胞癌（HCC）和肝硬化是全球范围内重要的死亡原因。持久性乙型肝炎病毒（HBV）感染是这些疾病的主要原因。病毒的基础核心启动子（BCP）（A1762T和G1764A）和前核心（pre-C）（G1896A）区的双重突变与进展为HCC有关。当前的研究旨在开发一种简单的方法来筛选和检测HBV携带者的BCP和pre-C突变。我们已经开发并验证了寡核苷酸连接测定法（OLA），以检测HBV核心基因中的点突变。我们将OLA方法应用于从冈比亚肝癌研究（GLCS）招募的HBV感染携带者的样本中，这些样本包括无症状HBsAg携带者，肝硬化患者和HCC患者。我们观察到BCP和pre-C突变的OLA和DNA测序之间的一致性分别为89.3％和95.8％。 OLA在仅检测到单个显性菌株的条件下，检测了单株感染以及野生型和突变型病毒混合物感染中的突变。在进行性肝病（肝硬化/ HCC）的患者中检测到BCP突变的比例为75.7％，而在无症状携带者中为47.6％，而在进行性肝病患者中，有34.5％的患者为C-pre突变，在无症状HBsAg携带者中为47.6％。 BCP突变的存在与晚期肝病之间存在显着关联。总之，OLA是一种用于检测前C和BCP突变的简单，经济且可靠的检测方法。它的应用可以改善HBV流行地区的诊断和临床护理。

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