We examined the reliability of the method in clinical samples, and found it could detect the HBV subtypes and drug resistance mutations in 80 clinical HBV samples with low HBV DNA levels ranging from 20 to 200 IU/mL. These primers could efficiently amplify the RT region of HBV virus at low DNA levels by directly sequencing the resulting PCR products, and mapping with the reference sequence made it possible to clearly obtain the HBV subtypes and identify the resistance mutations in the samples with HBV DNA level as low as 20 IU/mL. The novel, common, and universal primers were identified by alignment of RT region of all the HBV DNA sequences in databases. Also, sensitive detection of HBV antiviral drug resistance mutations is essential for monitoring therapy response.Īsensitive direct sequencing method for genotyping and the drug resistance mutation detection of low levels of HBV DNA in patients’ plasma is developed by PCR amplification of the DNA with novel universal primers. HBV (hepatitis B virus) genotyping is important in determining the clinical manifestation of disease and treatment response, particularly, in patients with low viral loads. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial License 4.0 (CCBY-NC), where it is permissible to download, share, remix, transform, and buildup the work provided it is properly cited. This work was financially supported by the Hubei Provincial Natural Science Foundation of China (2010CDB06903), National Natural Science Foundation of China (81000771), National Key Basic Research Program of China-973 Program (2012CB526706), and the National Natural Science Foundation of China (81271694).Īll the authors in the manuscript have no conflict of interests to declare. YT, BL, and HL contributed equally to this work. When transformations of marine organic matter are considered, differences in community composition and their different abilities to access organic matter should be taken into account.BClinical Molecular Diagnostic Center, Renmin Hospital of Wuhan UniversityĬDepartment of Pathology Affiliated Tianyou Hospital of Wuhan University of Science and Technology, WuhanĭEastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, ChinaĮPennsylvania State University College of Medicine and Hershey Medical center, Hershey, PAįNew York University College of Arts and Science, New York, NY.Ĭorrespondence: Yan Li, Department of Clinical Laboratory, Renmin Hospital of Wuhan University, 99 Ziyang Road of Wuchang District, Wuhan 430060, China (e-mail: ) Chunhua Song, Pennsylvania State University College of Medicine, Penn State Hershey Children's Hospital, PO Box 850, 500 University Drive, Hershey, PA 17033 (e-mail: ).Ībbreviations: ETV = entecavir, HBV = hepatitis B virus, HCC = hepatocellular carcinoma, IFN-a = interferon-alpha, LAM = lamivudine, LdT = telbivudine, NAs = nucleoside analogs, qPCR = quantitative real-time PCR. The greater number of phylum- and subphylum-level lineages and operational taxonomic units in sediments than in seawater samples may reflect the necessity of a wider range of enzymatic capabilities and strategies to access organic matter that has already been degraded during passage through the water column.
Thus, the broader enzymatic capabilities of the sedimentary microbial communities may result from the compositional differences between seawater and sedimentary microbial communities, rather than from gene expression differences among compositionally similar communities. To compare bacterial communities, 16S rRNA gene clone libraries were constructed from the same seawater and sediment samples they diverged strongly in composition. In seawater, in contrast, only 5 of the 7 polysaccharides and 2 of the 3 algal extracts were hydrolyzed, and hydrolysis rates in surface and deepwater were virtually identical. Sedimentary microbial communities hydrolyzed all of the fluorescently labeled polysaccharide and algal extracts, in most cases at higher rates in subsurface than surface sediments.
Patterns of enzyme activities differed between seawater and sediments, not just quantitatively, in accordance with higher cell numbers in sediments, but also in their more diversified enzyme spectrum. communities to initiate organic matter degradation, we measured the extracellular enzymatic hydrolysis rates of 10 substrates (polysaccharides and algal extracts) in surface seawater and bottom water as well as in surface and anoxic sediments of an Arctic fjord. To compare the abilities of seawater and sedimentary microbial. Although carbon cycling and preservation depend critically on the capabilities of these microbial communities, their compositions and capabilities have seldom been examined simultaneously at the same site. Heterotrophic microbial communities in seawater and sediments metabolize much of the organic carbon produced in the ocean.
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