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Mendeley readers
Attention Score in Context
| Title |
A novel method for detection of HBVcccDNA in hepatocytes using rolling circle amplification combined with in situ PCR
|
|---|---|
| Published in |
BMC Infectious Diseases, December 2014
|
| DOI | 10.1186/s12879-014-0608-y |
| Pubmed ID | |
| Authors |
Yanwei Zhong, Shuangye Hu, Chen Xu, Yulai Zhao, Dongping Xu, Yanqing Zhao, Jingmin Zhao, Zhibin Li, Xiuchang Zhang, Hongfei Zhang, Jin Li |
| Abstract |
BackgroundIntrahepatic hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) is the original template for HBV replication. The persistence of cccDNA is responsible for the recurrence of HBV infection. The detection of cccDNA can help the development of new antiviral drugs against HBV replication links, and reduce the resistance and recurrence as well as to discover extrahepatic HBV infection. In situ polymerase chain reaction (IS-PCR) can be used to determine the distribution and localization of cccDNA in liver tissues, but it is hampered by its low sensitivity and specificity. We developed a novel method to detect HBV cccDNA using rolling circle amplification (RCA) combined with IS-PCR.MethodsBiopsy liver tissues were obtained from 26 patients with HBV infection, including 10 chronic hepatitis B (CHB), 6 liver cirrhosis (LC) and 10 hepatocellular carcinoma (HCC) patients. Four pairs of primers were designed to mediating RCA for the first round amplification of HBV cccDNA specifically. The liver tissue sections from patients were treated by plasmid-safe ATP-dependent DNase (PSAD) prior to RCA. After RCA, HBV cccDNA was further amplified by a pair of selective primers labeled digoxigenin that target the gap region between the two direct repeat regions (DR1 and DR2) of the virus via IS-PCR.ResultsHBVcccDNA was expressed and located in hepatocyte nucleus in 19 patients (73.07%). Compared with the IS-PCR, the introduction of RCA increase the limit of detection. RCA combined with IS-PCR yielded strong positive signals in HCC liver tissue in spite of low copy number cccDNA (2 copies of target sequence per cell), meanwhile, no positive signal was detected via negative control.ConclusionsRCA combined with IS-PCR is an effective and practicable method which could detect the presence of low copy number of cccDNA sensitively and specifically, and reflect the relationship between cccDNA expression level and liver tissue pathological characteristics. |
X Demographics
The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 44 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 44 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 12 | 27% |
| Student > Ph. D. Student | 7 | 16% |
| Student > Bachelor | 5 | 11% |
| Other | 4 | 9% |
| Student > Master | 2 | 5% |
| Other | 5 | 11% |
| Unknown | 9 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 12 | 27% |
| Agricultural and Biological Sciences | 8 | 18% |
| Medicine and Dentistry | 6 | 14% |
| Immunology and Microbiology | 3 | 7% |
| Chemistry | 2 | 5% |
| Other | 2 | 5% |
| Unknown | 11 | 25% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 04 December 2014.
All research outputs
#20,245,139
of 22,772,779 outputs
Outputs from BMC Infectious Diseases
#6,459
of 7,668 outputs
Outputs of similar age
#302,268
of 360,895 outputs
Outputs of similar age from BMC Infectious Diseases
#165
of 197 outputs
Altmetric has tracked 22,772,779 research outputs across all sources so far. This one is in the 1st percentile – i.e., 1% of other outputs scored the same or lower than it.
So far Altmetric has tracked 7,668 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 9.6. This one is in the 1st percentile – i.e., 1% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 360,895 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 197 others from the same source and published within six weeks on either side of this one. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.