肝胆相照论坛
标题: 使用ddPCR高度灵敏地检测乙型肝炎 [打印本页]
作者: StephenW 时间: 2018-4-13 12:13 标题: 使用ddPCR高度灵敏地检测乙型肝炎
GEN News Highlights More »
April 12, 2018
Highly Sensitive Detection of Hepatitis B Using ddPCR 
Hepatitis B virus (HBV) closed circular DNA (cccDNA) detection is of major clinical significance. HBV entry into the nucleus of liver cells (hepatocytes) stimulates cccDNA to create HBV RNAs that give rise to virus offspring. Measurement of cccDNA levels can help clinicians determine the level of HBV infection. [Song-Mei Liu/Zhongnan Hospital of Wuhan University]
- Developing new, more sensitive assays to detect chronic infections, like those from the hepatitis B virus (HBV), is imperative to when trying to manage associated diseases such as cirrhosis and hepatocellular carcinoma (HCC). Now, a new study from investigators at the Center for Gene Diagnosis in the Zhongnan Hospital of Wuhan University, China, describes a new HBV assay that offers advantages over currently used methods because it can detect closed circular DNA (cccDNA) in serum, single cells, and preserved tissue samples. Findings from the new study were published today in The Journal of Molecular Diagnostics, in an article entitled “A Highly Sensitive and Robust Method for Hepatitis B Virus Covalently Closed Circular DNA Detection in Single Cells and Serum.”
"The development of HCC is strongly associated with HBV. Recently, several new antiviral strategies targeting cccDNA have been established to improve HBV clearance,” explained lead study investigator Song-Mei Liu, M.D., Ph.D., chair and vice professor at the Center for Gene Diagnosis. “It is of great clinical significance to provide an accurate and sensitive approach for cccDNA detection. With this method, more and more patients with chronic HBV will have precision treatment available to prevent or delay HCC occurrence, and HCC in patients could be diagnosed at an earlier stage.”
According to the World Health Organization, 257 million individuals worldwide are infected with HBV, and in 2015 HBV resulted in 887,000 deaths, primarily due to liver complications. Chronic HBV infection can be treated with medications such as tenofovir or entecavir. In 2015, only 9% of individuals living with HBV infection were aware they were infected.
The new assay utilizes droplet digital PCR (ddPCR), which is highly sensitive and accurate for detecting trace molecules such as viruses. Using the ddPCR method, the researchers were able to detect cccDNA in serum, single cells, and formalin-fixed, paraffin-embedded tumor issues. Southern blotting, currently considered the gold standard for this purpose, and other assays have important limitations such as decreased sensitivity compared to ddPCR in detecting low copy numbers of cccDNA. Moreover, traditional methods often require invasive techniques like obtaining liver tissue through biopsy.
"Compared to liver biopsy, serum can be obtained noninvasively, is widely used for clinical diagnostic purposes, and has a homogeneous cccDNA distribution,” Dr. Liu noted. “The assay also improves the limit of detection for cccDNA.”
The new ddPCR method was useful in identifying patients that might be harboring HCC. The research team found that almost 90% of 68 HCC patients were cccDNA-positive compared to 53% of 79 non-HCC patients. Serum cccDNA copy number was found to be higher in HCC patients compared to non-HCC patients. Combined analysis of serum cccDNA and HBV-DNA distinguished HCC patients from non-HCC patients. "This implicates cccDNA as a risk factor for HCC," Dr. Liu remarked.
Additionally, the scientists were able to confirm that serum cccDNA was positively correlated with levels of cccDNA measured in liver samples. "Serum cccDNA is indeed a much better and useful diagnostic marker than intrahepatic cccDNA," Dr. Liu concluded.
Recently, several new antiviral strategies targeting cccDNA have been found to improve HBV clearance. Therefore, providing an accurate and sensitive approach for cccDNA detection is of great clinical significance for earlier diagnosis and HCC prediction, targeted treatment, and evaluating treatment efficacy.
作者: StephenW 时间: 2018-4-13 12:14
GEN新闻聚焦
更多 ”
2018年4月12日
使用ddPCR高度灵敏地检测乙型肝炎
乙型肝炎病毒（HBV）闭环DNA（cccDNA）检测具有重要的临床意义。 HBV进入肝细胞核（肝细胞）刺激cccDNA产生HBV RNAs，产生病毒后代。 cccDNA水平的测量可以帮助临床医生确定HBV感染的水平。 [刘松梅/武汉大学中南医院]
当试图控制相关疾病如肝硬化和肝细胞癌（HCC）时，开发新的，更灵敏的检测方法来检测慢性感染，如来自乙型肝炎病毒（HBV）的慢性感染。现在，中国武汉大学中南医院基因诊断中心的一项新研究描述了一种新的HBV检测方法，它比目前使用的方法具有优势，因为它可以检测血清中的闭合环状DNA（cccDNA），单细胞和保存的组织样本。这项新研究的发现今天在“分子诊断学杂志”上发表，题为“高灵敏度和稳健的乙肝病毒共价闭合圆形DNA检测单细胞和血清方法”的文章。
研究负责人Song-Mei Liu博士说：“HCC的发展与HBV密切相关，近来，已经建立了几种针对cccDNA的新型抗病毒策略来改善HBV清除率。 “基因诊断中心”提供准确而敏感的cccDNA检测方法具有重大的临床意义，用这种方法，越来越多的慢性HBV患者将有精确的治疗方法来预防或延迟HCC的发生，患者中的HCC可以在早期进行诊断。“
据世界卫生组织统计，全球有2.57亿人感染了HBV，2015年HBV导致887,000人死亡，主要是由于肝脏并发症。慢性HBV感染可以用替诺福韦或恩替卡韦等药物治疗。在2015年，只有9％的感染乙肝病毒的人知道他们受到感染。
新的分析方法采用液滴数字PCR（ddPCR），它对检测痕量分子如病毒具有高度的敏感性和准确性。使用ddPCR方法，研究人员能够检测血清中的cccDNA，单细胞和福尔马林固定的石蜡包埋肿瘤问题。 Southern印迹法目前被认为是用于此目的的金标准，其他测定法具有重要限制，例如与检测cccDNA的低拷贝数的ddPCR相比灵敏度降低。此外，传统方法通常需要侵入性技术，如通过活检获取肝脏组织。
“与肝活检相比，血清可以非侵袭性地获得，被广泛用于临床诊断目的，并且具有均一的cccDNA分布，”刘博士指出，“该检测方法还可以提高cccDNA检测的极限。”
新的ddPCR方法可用于鉴定可能携带HCC的患者。该研究小组发现，68例HCC患者中近90％为cccDNA阳性，而79例非HCC患者中有53％为cccDNA阳性。发现血清cccDNA拷贝数在HCC患者中高于非HCC患者。血清cccDNA和HBV-DNA联合分析鉴别非HCC患者的HCC患者。 “这暗示cccDNA是HCC的危险因素，”刘博士评论道。
此外，科学家们能够证实血清cccDNA与肝脏样本中测得的cccDNA水平正相关。 “血清cccDNA确实是一种比肝内cccDNA更好，更有用的诊断标记，”刘博士总结说。
最近，已发现几种靶向cccDNA的新型抗病毒策略可改善HBV清除率。因此，为早期诊断和HCC预测，靶向治疗和评估治疗效果提供准确而敏感的cccDNA检测方法，具有重要的临床意义。
作者: antiHBVren 时间: 2018-4-13 12:45
这个不错，之前的cccDNA的精度太低。
1. 新的分析方法ddPCR 对检测痕量分子如病毒具有高度的敏感性和准确性。可以替代 活检获取肝脏组织；
2. 新的分析方法ddPCR还可以提高cccDNA检测的极限。
3. 新的ddPCR方法可用于鉴定可能携带HCC的患者。 这个直接能筛查出HCC高危患者
4. 因为已经能够证实血清cccDNA与肝脏样本中测得的cccDNA水平正相关，血清cccDNA确实是一种比肝内cccDNA更好
5. 为已发现几种靶向cccDNA的新型抗病毒策略，为早期诊断和HCC预测，靶向治疗和评估治疗效果提供准确而敏感的cccDNA检测方法
作者: newchinabok 时间: 2018-4-13 13:36
英特纳雄耐尔就一定要实现
作者: antiHBVren 时间: 2018-4-16 10:22
回复 newchinabok 的帖子
这个必须的！
希望还是要有的
| 欢迎光临 肝胆相照论坛 (http://hbvhbv.info/forum/) |
Powered by Discuz! X1.5 |