Use of real-time PCR and fluorimetry to detect lamivudine resistance-associated mutations in hepatitis B virus.


:Very rapid amplification of DNA by PCR in small volumes can be continuously monitored by the detection of the binding of probes with a rapid cycler with built-in fluorometric detection. Primers were designed to amplify approximately 100 bp of the polymerase gene of hepatitis B virus (HBV) spanning codon 550, where mutations associated with resistance to lamivudine invariably occur. Four hybridization probes were synthesized: one was 3' labelled with fluorescein and hybridized upstream of codon 550. The others were 5' labelled with Cy5 and 3' labelled with biotin and spanned codon 550. The Cy5-labelled oligonucleotides contained either wild-type (ATG) or mutant (GTG or ATT) sequences. A Cy5-labelled probe and either the fluorescein-labelled probe or Sybr Green 1 (a compound that fluoresces when bound to double-stranded DNA) were included in each PCR. After completion of the amplification by using a LightCycler (Idaho Technology), the temperature at which the Cy5 probe melted from the product was determined in a melt program that took ca. 3 min. Pre- and posttreatment samples from eight patients (five chronic and three transplant) who failed lamivudine treatment were amplified, and the presence of mutations in codon 550 was determined by ABI sequencing and by using the LightCycler; in some cases PCR products were also cloned, and multiple clones were sequenced. Concordant results were obtained in all cases. We found the LightCycler to be better at resolving the sequences of genomic mixtures; for example, two samples showed a sequence at codon 550 of (A/G)T(G/T), which was found by fluorimetry to be mixtures of GTG and ATT but no ATG, and this finding was confirmed by the sequencing of clones. However, this approach was not more sensitive than population sequencing for the detection of the presence of mixtures. Overall, this pilot study has demonstrated an approach that could be an extremely rapid and economical method for the detection of lamivudine resistance-associated mutations in HBV.


Cane PA,Cook P,Ratcliffe D,Mutimer D,Pillay D




Has Abstract


1999-07-01 00:00:00












  • Rapid selection of organisms with increasing resistance on subinhibitory concentrations of norfloxacin in agar.

    abstract::Serial passage of Pseudomonas aeruginosa ATCC 27853 or Escherichia coli ATCC 25922 on agar with subinhibitory concentrations of norfloxacin rapidly produced isolates with minimal inhibitory concentrations (MICs) of norfloxacin up to 512-fold higher than that for the original strain. Although MICs of seven unrelated an...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Tenney JH,Maack RW,Chippendale GR

    更新日期:1983-01-01 00:00:00

  • Effects of azithromycin in combination with vancomycin, daptomycin, fosfomycin, tigecycline, and ceftriaxone on Staphylococcus epidermidis biofilms.

    abstract::Staphylococcal biofilms on surgical implants are the underlying cause of a lack of response to antimicrobial treatment. We investigated the effects of vancomycin (VAN), daptomycin (DAP), fosfomycin (FOS), tigecycline (TGC), and ceftriaxone (CRX), alone and in combination with azithromycin (AZI), on established biofilm...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Presterl E,Hajdu S,Lassnigg AM,Hirschl AM,Holinka J,Graninger W

    更新日期:2009-08-01 00:00:00

  • In vitro and in vivo pharmacokinetics-pharmacodynamics of GV143253A, a novel trinem.

    abstract::The aim of the present study was to characterize the pharmacokinetic-pharmacodynamic relationship of GV143253A, a novel trinem anti-methicillin-resistant Staphylococcus aureus (MRSA) agent active against gram-positive cocci, including multidrug-resistant clinical isolates. An in vitro pharmacodynamic study with methic...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Ferrari L,Iavarone L,Braggio S,Di Modugno E

    更新日期:2003-02-01 00:00:00

  • Safety, tolerability, and pharmacokinetics of PA-824 in healthy subjects.

    abstract::PA-824 is a novel antibacterial agent that has shown in vitro activity against both drug-sensitive and drug-resistant Mycobacterium tuberculosis. The compound's MIC is between 0.015 and 0.25 microg/ml for drug-sensitive strains and between 0.03 and 0.53 microg/ml for drug-resistant strains. In addition, it is active a...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章,随机对照试验


    authors: Ginsberg AM,Laurenzi MW,Rouse DJ,Whitney KD,Spigelman MK

    更新日期:2009-09-01 00:00:00

  • Pharmacokinetics and safety of a new parenteral carbapenem antibiotic, biapenem (L-627), in elderly subjects.

    abstract::The pharmacokinetics and tolerability of a new parenteral carbapenem antibiotic, biapenem (L-627), were studied in healthy elderly volunteers aged 65 to 74 years (71.6 +/- 2.7 years [mean +/- standard deviation], n = 5; group B) and > or = 75 years (77.8 +/- 1.9 years, n = 5; group C), following single intravenous dos...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Kozawa O,Uematsu T,Matsuno H,Niwa M,Takiguchi Y,Matsumoto S,Minamoto M,Niida Y,Yokokawa M,Nagashima S,Kanamaru M

    更新日期:1998-06-01 00:00:00

  • Whole-genome sequencing of gentamicin-resistant Campylobacter coli isolated from U.S. retail meats reveals novel plasmid-mediated aminoglycoside resistance genes.

    abstract::Aminoglycoside resistance in Campylobacter has been routinely monitored in the United States in clinical isolates since 1996 and in retail meats since 2002. Gentamicin resistance first appeared in a single human isolate of Campylobacter coli in 2000 and in a single chicken meat isolate in 2007, after which it increase...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Chen Y,Mukherjee S,Hoffmann M,Kotewicz ML,Young S,Abbott J,Luo Y,Davidson MK,Allard M,McDermott P,Zhao S

    更新日期:2013-11-01 00:00:00

  • Elimination of plasmid pMG110 from Escherichia coli by novobiocin and other inhibitors of DNA gyrase.

    abstract::The ability of novobiocin to eliminate (cure) the wild-type plasmid pMG110 from Escherichia coli has been compared with that of other inhibitors of the gyrase B subunit and of the gyrase A subunit. Novobiocin eliminated pMG110 , producing over 99% plasmid loss at concentrations two- to eightfold below the MIC for bact...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Hooper DC,Wolfson JS,McHugh GL,Swartz MD,Tung C,Swartz MN

    更新日期:1984-05-01 00:00:00

  • Differences in the growth of Aspergillus fumigatus on cycloheximide media at three temperatures.

    abstract::Cycloheximide (up to 0.4 g/liter) was significantly more inhibitory to the growth of three isolates of Aspergillus fumigatus at 23 C than at 37 or 45 C. Preincubation of the media for 7 days at 45 C did not alter the inhibitory effect of the cycloheximide at 23 C. Neither mutation nor adaptation in the fungus seems to...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Berger CL,Merz WG,Silva-Hutner M

    更新日期:1973-10-01 00:00:00

  • Pharmacokinetics of levofloxacin and ciprofloxacin during continuous renal replacement therapy in critically ill patients.

    abstract::The pharmacokinetics of intravenously administered levofloxacin and ciprofloxacin were studied in intensive care unit patients during continuous venovenous hemofiltration (CVVH; four patients received levofloxacin, and five received ciprofloxacin) or hemodiafiltration (CVVHDF; six patients received levofloxacin, and f...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Malone RS,Fish DN,Abraham E,Teitelbaum I

    更新日期:2001-10-01 00:00:00

  • Penetration of fleroxacin into breast milk and pharmacokinetics in lactating women.

    abstract::Fleroxacin was administered to seven lactating women as a single oral dose of 400 mg. Plasma, urine, and milk samples were collected for up to 48 h after administration. Drug concentrations were determined by a reversed-phase high-pressure liquid chromatography method and were used for the pharmacokinetic evaluation. ...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Dan M,Weidekamm E,Sagiv R,Portmann R,Zakut H

    更新日期:1993-02-01 00:00:00

  • Retrograde Trafficking Inhibitor of Shiga Toxins Reduces Morbidity and Mortality of Mice Infected with Enterohemorrhagic Escherichia coli.

    abstract::The most deadly outbreak of Escherichia coli O104:H4 occurred in Europe in 2011. Here, we evaluated the effects of the retrograde trafficking inhibitor Retro-2(cycl) in a murine model of E. coli O104:H4 infection. Systemic treatment with Retro-2(cycl) significantly reduced body weight loss and improved clinical scores...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Secher T,Shima A,Hinsinger K,Cintrat JC,Johannes L,Barbier J,Gillet D,Oswald E

    更新日期:2015-08-01 00:00:00

  • Echinocandin susceptibility testing of Candida species: comparison of EUCAST EDef 7.1, CLSI M27-A3, Etest, disk diffusion, and agar dilution methods with RPMI and isosensitest media.

    abstract::This study compared nine susceptibility testing methods and 12 endpoints for anidulafungin, caspofungin, and micafungin with the same collection of blinded FKS hot spot mutant (n = 29) and wild-type isolates (n = 94). The susceptibility tests included EUCAST Edef 7.1, agar dilution, Etest, and disk diffusion with RPMI...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Arendrup MC,Garcia-Effron G,Lass-Flörl C,Lopez AG,Rodriguez-Tudela JL,Cuenca-Estrella M,Perlin DS

    更新日期:2010-01-01 00:00:00

  • Novel functions of an iron-sulfur flavoprotein from Trichomonas vaginalis hydrogenosomes.

    abstract::Iron-sulfur flavoproteins (Isf) are flavin mononucleotide (FMN)- and FeS cluster-containing proteins commonly encountered in anaerobic prokaryotes. However, with the exception of Isf from Methanosarcina thermophila, which participates in oxidative stress management by removing oxygen and hydrogen peroxide, none of the...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Smutná T,Pilarová K,Tarábek J,Tachezy J,Hrdý I

    更新日期:2014-06-01 00:00:00

  • Comparative antimicrobial activity of O-demethylfortimicin A, a derivative of fortimicin A.

    abstract::The in vitro antimicrobial activity of O-demethylfortimicin A (ODMF), a derivative of fortimicin A, was compared with those of fortimicin A and gentamicin against a spectrum of 256 organisms. All three antibiotics were active in low concentrations against all strains of Enterobacteriaceae, Acinetobacter sp., and Staph...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Girolami RL,Stamm JM

    更新日期:1980-11-01 00:00:00

  • Activity of telithromycin (HMR 3647) against anaerobic bacteria compared to those of eight other agents by time-kill methodology.

    abstract::Time-kill studies examined the activities of telithromycin (HMR 3647), erythromycin A, azithromycin, clarithromycin, roxithromycin, clindamycin, pristinamycin, amoxicillin-clavulanate, and metronidazole against 11 gram-positive and gram-negative anaerobic bacteria. Time-kill studies were carried out with the addition ...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Credito KL,Ednie LM,Jacobs MR,Appelbaum PC

    更新日期:1999-08-01 00:00:00

  • Effect of cimetidine on the disposition of rimantadine in healthy subjects.

    abstract::Twenty-three healthy male and female subjects received single 100-mg oral doses of rimantadine hydrochloride on two occasions in an open-label, sequential design with a 6-day washout between doses. The first dose of rimantadine was administered alone, and the second dose was administered concomitantly with cimetidine ...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Holazo AA,Choma N,Brown SY,Lee LF,Wills RJ

    更新日期:1989-06-01 00:00:00

  • Ultra-performance liquid chromatography mass spectrometry and sensitive bioassay methods for quantification of posaconazole plasma concentrations after oral dosing.

    abstract::Posaconazole (POS) is a new antifungal agent for prevention and therapy of mycoses in immunocompromised patients. Variable POS pharmacokinetics after oral dosing may influence efficacy: a trough threshold of 0.5 μg/ml has been recently proposed. Measurement of POS plasma concentrations by complex chromatographic techn...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Rochat B,Pascual A,Pesse B,Lamoth F,Sanglard D,Decosterd LA,Bille J,Marchetti O

    更新日期:2010-12-01 00:00:00

  • OXA-14, another extended-spectrum variant of OXA-10 (PSE-2) beta-lactamase from Pseudomonas aeruginosa.

    abstract::Pseudomonas aeruginosa 455, isolated in Ankara, Turkey, produced a pI 6.2 beta-lactamase determined by plasmid pMLH53 and resisted all beta-lactams except carbapenems. This beta-lactamase, named OXA-14, corresponded to OXA-10 (PSE-2) except that aspartate replaced glycine at position 157 and thus is intermediate betwe...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Danel F,Hall LM,Gur D,Livermore DM

    更新日期:1995-08-01 00:00:00

  • Distribution of human immunodeficiency virus type 1 protease and reverse transcriptase mutation patterns in 4,183 persons undergoing genotypic resistance testing.

    abstract::In a sample of 6,156 sequences from 4,183 persons, the top 30 patterns of protease inhibitor, nucleoside reverse transcriptase (RT) inhibitor, and nonnucleoside RT inhibitor mutations accounted for 55, 46, and 66%, respectively, of sequences with drug resistance mutations. Characterization of the phenotypic and clinic...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Rhee SY,Liu T,Ravela J,Gonzales MJ,Shafer RW

    更新日期:2004-08-01 00:00:00

  • Screening and Characterization of Multidrug-Resistant Gram-Negative Bacteria from a Remote African Area, São Tomé and Príncipe.

    abstract::The occurrence of resistance to last-resort antibiotics was evaluated among Enterobacteriaceae isolates recovered from hospitalized children in a remote African archipelago, São Tomé and Príncipe, where there is limited access to those antibiotics. Fifty patients were screened for colonization by carbapenem-, pan-amin...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Poirel L,Aires-de-Sousa M,Kudyba P,Kieffer N,Nordmann P

    更新日期:2018-08-27 00:00:00

  • Mode of action of the biotin antimetabolites actithiazic acid and alpha-methyldethiobiotin.

    abstract::Actithiazic acid and alpha-methyldethiobiotin inhibited the conversion of dethiobiotin to biotin resting-cell suspensions of Escherichia coli. The concentrations which effected 50% inhibition were 0.45 and 1.1 microM for actithiazic acid and alpha-methyldethiobiotin, respectively. Cells grown in low concentrations of ...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Eisenberg MA,Hsiung SC

    更新日期:1982-01-01 00:00:00

  • Molecular Analysis of a blaIMP-1-Harboring Class 3 Integron in Multidrug-Resistant Pseudomonas fulva.

    abstract::A multidrug-resistant (MDR) Pseudomonas fulva strain was isolated in 2006 from a urine sample. The isolate harbored the blaIMP-1 gene, which was located in a chromosomal Tn402-like class 3 integron as a gene cassette array of aacA31-fosE-blaIMP-1 Two mutations in gyrA and one mutation in parC were detected in quinolon...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Yamamoto M,Matsumura Y,Gomi R,Matsuda T,Nakano S,Nagao M,Tanaka M,Ichiyama S

    更新日期:2018-07-27 00:00:00

  • In vitro and in vivo inhibition of erythrocytic development of malarial parasites by docetaxel.

    abstract::The stage-dependent susceptibility of Plasmodium falciparum to a short exposure to docetaxel (Taxotere) was evaluated by subjecting ring-infected, trophozoite-infected, and schizont-infected erythrocytes to a 5-h exposure to various concentrations of the drug. The schizont stage was shown to be the most sensitive stag...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Sinou V,Grellier P,Schrevel J

    更新日期:1996-02-01 00:00:00

  • Plasmid-mediated high-level resistance to aminoglycosides in Enterobacteriaceae due to 16S rRNA methylation.

    abstract::A self-transferable plasmid of ca. 80 kb, pIP1204, conferred multiple-antibiotic resistance to Klebsiella pneumoniae BM4536, which was isolated from a urinary tract infection. Resistance to beta-lactams was due to the bla(TEM1) and bla(CTX-M) genes, resistance to trimethroprim was due to the dhfrXII gene, resistance t...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Galimand M,Courvalin P,Lambert T

    更新日期:2003-08-01 00:00:00

  • Comparative efficacies of lipid-complexed amphotericin B and liposomal amphotericin B against coccidioidal meningitis in rabbits.

    abstract::In separate previous studies, we have shown that lipid-complexed amphotericin B (Abelcet [ABLC]) and liposomal amphotericin B (AmBisome [AmBi]) are efficacious against coccidioidal meningitis in rabbits. Here, we compared ABLC and AmBi directly in a coccidioidal meningitis model. Male New Zealand White rabbits were in...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Clemons KV,Capilla J,Sobel RA,Martinez M,Tong AJ,Stevens DA

    更新日期:2009-05-01 00:00:00

  • Simple spectrophotometric assay for measuring protein binding of penem antibiotics to human serum.

    abstract::The binding of antibiotics to plasma (serum) proteins through hydrogen bonding can significantly influence the biological characteristics of these drugs. A rapid spectrophotometric assay has been developed that measures the level of free (unbound) penem antibiotic in serum ultrafiltrates. Whole human serum was adjuste...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Gootz TD,Subashi TA,Lindner DL

    更新日期:1988-02-01 00:00:00

  • Itraconazole resistance in Aspergillus fumigatus.

    abstract::Invasive aspergillosis is an increasingly frequent opportunistic infection in immunocompromised patients. Only two agents, amphotericin B and itraconazole, are licensed for therapy. Itraconazole acts through inhibition of a P-450 enzyme undertaking sterol 14alpha demethylation. In vitro resistance in Aspergillus fumig...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Denning DW,Venkateswarlu K,Oakley KL,Anderson MJ,Manning NJ,Stevens DA,Warnock DW,Kelly SL

    更新日期:1997-06-01 00:00:00

  • Efficacy and Safety of Pyronaridine-Artesunate plus Single-Dose Primaquine for Treatment of Uncomplicated Plasmodium falciparum Malaria in Eastern Cambodia.

    abstract::In Cambodia, multidrug-resistant Plasmodium falciparum undermines the treatment of uncomplicated malaria, and new therapeutic options are needed. Pyronaridine-artesunate has not previously been evaluated in eastern Cambodia. We conducted a single-arm, open-label, prospective study between July and December 2017 at the...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Leang R,Mairet-Khedim M,Chea H,Huy R,Khim N,Mey Bouth D,Dorina Bustos M,Ringwald P,Witkowski B

    更新日期:2019-02-26 00:00:00

  • Structure-absorption relationships of a series of 6-fluoroquinolones.

    abstract::The physicochemical constants and some structural parameters (topological, steric, and electronic) of eight third-generation monofluorate quinolones (six uncommercialized and two used clinically [ciprofloxacin and enrofloxacin]) were determined: pKa, intrinsic solubility (S0), chromatographic capacity factor, partitio...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Escribano E,Calpena AC,Garrigues TM,Freixas J,Domenech J,Moreno J

    更新日期:1997-09-01 00:00:00

  • Inactivation of Herpesvirus hominis types 1 and 2 by silver nitrate in vitro and in vivo.

    abstract::Herpes simplex virus (HSV) types 1 and 2 (two strains each) were inactivated at different rates in vitro by 40 muM AgNO(3). The inactivation of HSV type 1 strains was virtually complete in 10 to 15 min, whereas almost half of the infectivity of HSV type 2 strains survived this exposure. One strain of type 1 inoculated...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章


    authors: Coleman VR,Wilkie J,Levinson WE,Stevens T,Jawetz E

    更新日期:1973-09-01 00:00:00