Determination of HBV DNA viral load in Serum positive Hepatitis B patients using Quantitative Real Time PCR (qPCR)

Voke Toye, Emmanuel Afolayemi, Dorcas Akinuli, Alfred Akinlalu, Ijeoma Asilebo
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Abstract

Viral DNA quantification of Hepatitis B Virus (HBV) is crucial in treating and managing patients with chronic HBV disease. Although serological testing has contributed immensely to the disease diagnosis, molecular methods such as the quantitative Polymerase Chain Reaction (qPCR) are vital in viral load determination. This quantification technique is important for physicians to monitor the progress of antiviral therapy and determine future therapeutic options. In this study, we evaluated the detection efficiency of commercially available kits used in qPCR detection for the HBV viral load quantification. Serum cell-free circulating DNA of 37 patients that have been serologically confirmed positive for HBV and 13 healthy controls were extracted, and qPCR viral load determination was carried out. The results showed that only 13.51 % of the total HBV-positive samples were sufficiently high to be detected with viral load ranges from 0.87 – 1012 IU/mL. The high number of undetected cases 86.49 % may be attributed to the qPCR kit sensitivity as well as variations in the region of detection of viral genome found in the Nigerian population since these kits were designed for a different populace. This necessitates further studies that will validate the underlying mechanisms for the detection of the virus and see the need for indigenous detection kits to address the disease burden of HBV in Nigeria.
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定量实时PCR (qPCR)检测血清乙型肝炎阳性患者HBV DNA病毒载量
乙型肝炎病毒(HBV)的病毒DNA定量对慢性HBV疾病患者的治疗和管理至关重要。虽然血清学检测对疾病的诊断贡献巨大,但分子方法如定量聚合酶链反应(qPCR)在病毒载量测定中至关重要。这种量化技术对医生监测抗病毒治疗的进展和确定未来的治疗方案很重要。在这项研究中,我们评估了用于qPCR检测HBV病毒载量定量的市售试剂盒的检测效率。提取血清学证实HBV阳性的37例患者和13例健康对照者的血清无细胞循环DNA,进行qPCR病毒载量测定。结果显示,只有13.51%的hbv阳性样本的病毒载量在0.87 ~ 1012 IU/mL之间,达到了检测的高度。未检出病例的高数量(86.49%)可能归因于qPCR试剂盒的敏感性以及尼日利亚人群中发现的病毒基因组检测区域的差异,因为这些试剂盒是为不同的人群设计的。这需要进一步的研究,以验证病毒检测的基本机制,并认识到需要本土检测试剂盒来解决尼日利亚乙型肝炎病毒的疾病负担。
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