{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,28]],"date-time":"2025-09-28T11:16:19Z","timestamp":1759058179653},"reference-count":29,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2013,9,1]],"date-time":"2013-09-01T00:00:00Z","timestamp":1377993600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2013,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Resistance to antiviral treatment for chronic hepatitis B virus (HBV) has been associated with mutations in the HBV polymerase region. This study aimed at developing an ultrasensitive method for quantifying viral populations with all major HBV resistance-associated mutations, combining the amplification refractory mutation system real-time PCR (ARMS RT-PCR) with a molecular beacon using a LightCycler. The discriminatory ability of this method, the ARMS RT-PCR with molecular beacon assay, was 0.01 to 0.25% for the different HBV resistance-associated mutations, as determined by laboratory-synthesized wild-type (WT) and mutant (Mut) target sequences. The assay showed 100% sensitivity for the detection of mutant variants A181V, T184A, and N236T in samples from 41 chronically HBV-infected patients under antiviral therapy who had developed resistance-associated mutations detected by direct PCR Sanger sequencing. The ratio of mutant to wild-type viral populations (the Mut\/WT ratio) was &gt;1% in 38 (63.3%) of 60 samples from chronically HBV-infected nucleos(t)ide analogue-naive patients; combinations of mutations were also detected in half of these samples. The ARMS RT-PCR with molecular beacon assay achieved high sensitivity and discriminatory ability compared to the gold standard of direct PCR Sanger sequencing in identifying resistant viral populations in chronically HBV-infected patients receiving antiviral therapy. Apart from the dominant clones, other quasispecies were also quantified. In samples from chronically HBV-infected nucleos(t)ide analogue-naive patients, the assay proved to be a useful tool in detecting minor variant populations before the initiation of the treatment. These observations need further evaluation with prospective studies before they can be implemented in daily practice.<\/jats:p>","DOI":"10.1128\/jcm.00936-13","type":"journal-article","created":{"date-parts":[[2013,6,27]],"date-time":"2013-06-27T04:23:55Z","timestamp":1372307035000},"page":"2893-2900","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":12,"title":["Ultrasensitive Amplification Refractory Mutation System Real-Time PCR (ARMS RT-PCR) Assay for Detection of Minority Hepatitis B Virus-Resistant Strains in the Era of Personalized Medicine"],"prefix":"10.1128","volume":"51","author":[{"given":"Fotinie","family":"Ntziora","sequence":"first","affiliation":[{"name":"Department of Hygiene, Epidemiology, and Medical Statistics, University of Athens Medical School, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dimitrios","family":"Paraskevis","sequence":"additional","affiliation":[{"name":"Department of Hygiene, Epidemiology, and Medical Statistics, University of Athens Medical School, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catherine","family":"Haida","sequence":"additional","affiliation":[{"name":"Department of Hygiene, Epidemiology, and Medical Statistics, University of Athens Medical School, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emanuel","family":"Manesis","sequence":"additional","affiliation":[{"name":"Division of Internal Medicine, University of Athens Medical School, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George","family":"Papatheodoridis","sequence":"additional","affiliation":[{"name":"Second Department of Internal Medicine, Hippokration General Hospital, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Spilios","family":"Manolakopoulos","sequence":"additional","affiliation":[{"name":"Second Department of Internal Medicine, Hippokration General Hospital, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ioannis","family":"Elefsiniotis","sequence":"additional","affiliation":[{"name":"Hepatology Unit, Elena Venizelou Hospital, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Timokratis","family":"Karamitros","sequence":"additional","affiliation":[{"name":"Department of Hygiene, Epidemiology, and Medical Statistics, University of Athens Medical School, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexis","family":"Vassilakis","sequence":"additional","affiliation":[{"name":"Department of Hygiene, Epidemiology, and Medical Statistics, University of Athens Medical School, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angelos","family":"Hatzakis","sequence":"additional","affiliation":[{"name":"Department of Hygiene, Epidemiology, and Medical Statistics, University of Athens Medical School, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1055\/s-2006-939751"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/hep.21698"},{"key":"e_1_3_3_4_2","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1177\/135965350400900522","article-title":"Management of antiviral resistance in patients with chronic hepatitis B","volume":"9","author":"Locarnini S","year":"2004","unstructured":"LocarniniSHatzakisAHeathcoteJKeeffeEBLiangTJMutimerDPawlotskyJMZoulimF. 2004. 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