新测试可改善慢性乙型肝炎的诊断和管理

StephenW

                    New test offers improved diagnosis and management of chronic hepatitis B                                

COLD-PCR/FMCA enables simultaneous and more precise evaluation of HBV DNA, genotypes, and mutant DNA, as well as better assessment of infection status, disease progression, and response to treatment, reports The Journal of Molecular Diagnostics

               

Elsevier

   

            
  
  
                                    IMAGE: This is a schematic workflow for coamplification at lower denaturation temperature PCR (COLD-PCR)/fluorescence melting curve analysis (FMCA).       view more
   

Credit: The First Affliated Hospital of Fujian Medical University

          Philadelphia, October 10, 2019 - A report in the Journal of Molecular Diagnostics, published by Elsevier, describes a new and powerful laboratory tool that may improve the diagnosis and treatment of hepatitis B virus (HBV) infection. The technique can simultaneously assess several indicators important for optimal patient management.
        A team of scientists has developed a highly sensitive coamplification at lower denaturation temperature PCR (COLD-PCR) coupled with probe-based fluorescence melting curve analysis (FMCA) for precision diagnosis of chronic hepatitis B (CHB) patients. This novel tool is simple, stable, convenient, practical, inexpensive, and may be used routinely in the average hospital laboratory.
        "Guidelines have confirmed that dynamic monitoring of HBV DNA, genotypes, and reverse transcriptase (RT) mutant DNA is of great importance to assess infection status, predict disease progression, and judge treatment efficacy in HBV-infected patients," explained lead investigator Qishui Ou, PhD, Department of Laboratory Medicine, The First Af?liated Hospital of Fujian Medical University, the Gene Diagnostic Laboratory, Fujian Medical University, and the Fujian Key Laboratory of Laboratory Medicine, Fuzhou, China. "We believe COLD-PCR/FMCA provides a powerful laboratory tool for precise diagnosis and treatment of HBV-infected patients."
        Although a number of molecular methods have been developed for measuring these parameters, many are limited by poor sensitivity or inability to detect more than one mutation at a time. Others are too cumbersome or expensive for clinical use. "Our goal was to establish a more practical and inexpensive method with high sensitivity to detect genotype and RT mutations while detecting HBV DNA," noted Dr. Ou.
        Moreover, COLD-PCR/FMCA can detect HBV mutations at much lower concentrations than other techniques such as PCR/FMCA or PCR Sanger sequencing (1 percent vs. 10 percent vs. 20 percent, respectively). This new technique can also distinguish different phases of HBV infection according to the proportion and type of mutations as well as by detecting HBV DNA.
        The researchers also report that the genotype and mutation detected by COLD-PCR/FMCA may predict whether a patient will respond to antiviral therapy. Analysis of serum samples from 41 patients with CHB who were receiving entecavir revealed that the drug was most effective for patients with genotype B and those with a lower percentage of RT mutations at baseline or week 4.
        "Until now there have not been high-throughput approaches to detect HBV DNA, genotype, and RT mutations simultaneously. Therefore, it is necessary to establish a more practical and inexpensive method with high sensitivity to detect genotype and RT mutations while detecting HBV DNA. COLD-PCR/FMCA has that potential," said Dr. Ou.
        HBV infection affects the liver. According to the World Health Organization, as of 2015, 257 million individuals were living with and 887,000 died from HBV infection, usually as a result of cirrhosis (loss of liver cells and irreversible scarring of the liver) or liver cancer. In 2016, only about 10.5 percent of individuals infected with HBV were aware of their status, only a fraction of whom were receiving treatment. The virus is typically spread through contact with infected blood or bodily fluids. HBV can be prevented by vaccines that offer almost total protection against HBV infection.

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StephenW

新测试可改善慢性乙型肝炎的诊断和管理

COLD-PCR / FMCA能够同时更精确地评​​估HBV DNA，基因型和突变DNA，并更好地评估感染状况，疾病进展和对治疗的反应，《分子诊断杂志》报道

爱思唯尔
图片

图像：这是在较低变性温度PCR（COLD-PCR）/荧光熔解曲线分析（FMCA）时进行共扩增的示意性工作流程。查看更多

图片来源：福建医科大学附属第一医院

费城，2019年10月10日-Elsevier发表在《分子诊断学杂志》上的一份报告描述了一种新型而强大的实验室工具，可以改善乙肝病毒（HBV）感染的诊断和治疗。该技术可以同时评估对优化患者管理很重要的几个指标。

一组科学家在较低的变性温度PCR（COLD-PCR）结合基于探针的荧光熔解曲线分析（FMCA）上开发了一种高灵敏度的共扩增，可用于慢性乙型肝炎（CHB）患者的精确诊断。这种新颖的工具简单，稳定，方便，实用，廉价，并且可以在普通医院的实验室中常规使用。

首席研究员欧启水解释说：“指南已经证实，动态监测HBV DNA，基因型和逆转录酶（RT）突变DNA对于评估感染状况，预测疾病进展并判断对HBV感染的患者的治疗效果至关重要。”福建医科大学附属第一医院检验医学科，博士，福建医科大学基因诊断实验室，福建省检验医学重点实验室。 “我们相信COLD-PCR / FMCA为精确诊断和治疗HBV感染的患者提供了强大的实验室工具。”

尽管已开发出许多用于测量这些参数的分子方法，但由于灵敏度低或无法一次检测一个以上突变，许多方法受到了限制。其他的则太笨拙或太昂贵而无法用于临床。 Ou博士指出：“我们的目标是建立一种更实用，更便宜的方法，以高灵敏度检测基因型和RT突变，同时检测HBV DNA。”

此外，COLD-PCR / FMCA可以以比其他技术（例如PCR / FMCA或PCR Sanger测序）低得多的浓度检测HBV突变（分别为1％，10％和20％）。这项新技术还可以根据突变的比例和类型以及检测HBV DNA来区分HBV感染的不同阶段。

研究人员还报告说，通过COLD-PCR / FMCA检测到的基因型和突变可能预测患者是否会对抗病毒治疗产生反应。对接受恩替卡韦的41例CHB患者的血清样本的分析表明，该药物对基因型B和基线或第4周RT突变百分比较低的患者最有效。

“到现在为止，还没有高通量的方法可以同时检测HBV DNA，基因型和RT突变。因此，有必要建立一种更实用，更便宜的方法，以高灵敏度检测HBV DNA的同时检测基因型和RT突变。 COLD-PCR / FMCA具有这种潜力，” Ou博士说。

HBV感染会影响肝脏。根据世界卫生组织的数据，截止到2015年，2.57亿人患有HBV感染，其中88.7万人死于HBV感染，通常是由于肝硬化（肝细胞丢失和不可逆转的肝脏瘢痕形成）或肝癌引起的。 2016年，只有大约10.5％的HBV感染者知道自己的状况，只有一小部分正在接受治疗。该病毒通常通过与感染的血液或体液接触而传播。可以通过提供几乎完全抗HBV感染保护的疫苗来预防HBV。

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