Abstract:
:Fluorescent and colorimetric reporter genes are valuable tools for drug screening models, since microscopy is labor intensive and subject to observer variation. In this work, we propose a fluorimetric method for drug screening using red fluorescent parasites. Fluorescent Leishmania amazonensis were developed after transfection with integration plasmids containing either red (RFP) or green fluorescent protein (GFP) genes. After transfection, wild-type (LaWT) and transfected (LaGFP and LaRFP) parasites were subjected to flow cytometry, macrophage infection, and tests of susceptibility to current antileishmanial agents and propranolol derivatives previously shown to be active against Trypanosoma cruzi. Flow cytometry analysis discriminated LaWT from LaRFP and LaGFP parasites, without affecting cell size or granulosity. With microscopy, transfection with antibiotic resistant genes was not shown to affect macrophage infectivity and susceptibility to amphotericin B and propranolol derivatives. Retention of fluorescence remained in the intracellular amastigotes in both LaGFP and LaRFP transfectants. However, detection of intracellular RFP parasites was only achieved in the fluorimeter. Murine BALB/c macrophages were infected with LaRFP parasites, exposed to standard (meglumine antimoniate, amphotericin B, Miltefosine, and allopurinol) and tested molecules. Although it was possible to determine IC(50) values for 4 propranolol derivatives (1, 2b, 3, and 4b), all compounds were considered inactive. This study is the first to develop a fluorimetric drug screening test for L. amazonensis RFP. The fluorimetric test was comparable to microscopy with the advantage of being faster and not requiring manual counting.
journal_name
Diagn Microbiol Infect Disjournal_title
Diagnostic microbiology and infectious diseaseauthors
Rocha MN,Corrêa CM,Melo MN,Beverley SM,Martins-Filho OA,Madureira AP,Soares RPdoi
10.1016/j.diagmicrobio.2012.11.018subject
Has Abstractpub_date
2013-03-01 00:00:00pages
282-91issue
3eissn
0732-8893issn
1879-0070pii
S0732-8893(12)00501-9journal_volume
75pub_type
杂志文章abstract::The objective of the study is to compare the performance of conventional fluorescence microscopy (CFM) and light-emitting diode (LED) fluorescence microscopy (FM) for detection of acid-fast bacilli (AFB) in clinical samples. We included AFB smears, stained using the auramine O method and blindly examined with both CFM...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2013.03.023
更新日期:2013-07-01 00:00:00
abstract::A hydrogen-sulfide producing, citrate-positive strain of Escherichia coli isolated from urinary tract infection was found to be resistant to chloramphenicol (Cm), tetracycline (Tc), streptomycin (Sm), trimethoprim (Tmp), sulfamethoxazole (Smx), and cotrimoxazole (Tmp/Smx). The strain contained 7 plasmids of molecular ...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/0732-8893(96)00027-2
更新日期:1996-04-01 00:00:00
abstract::We conducted an open trial of oral ofloxacin, 400 mg q 12 hr, in the treatment of infections caused by Pseudomonas aeruginosa and other bacteria resistant to traditional oral antibiotics. There were 53 evaluable subjects, 30 having infection of the skin/skin structure, 13 of the respiratory tract, six of bone, and fou...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 临床试验,杂志文章
doi:10.1016/0732-8893(92)90085-8
更新日期:1992-07-01 00:00:00
abstract::Microarrays are widely used in high-throughput DNA and RNA hybridization tests and recently adopted to protein and small molecule interaction studies in basic research and diagnostics. Parallel detection of serum antibodies and antigens has several potential applications in epidemiologic research, vaccine development,...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2012.10.014
更新日期:2013-02-01 00:00:00
abstract::Numerous studies have been carried out to determine whether an Epstein-Barr virus (EBV) infection can be considered a risk factor for multiple sclerosis (MS), following the evidence of an increase in IgG response to nuclear antigen-1 (EBNA-1) in both serum and cerebrospinal fluid (CSF) from MS patients. However, the p...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2010.11.013
更新日期:2011-05-01 00:00:00
abstract:OBJECTIVES:We assessed the analytical and clinical performance of the Lumipulse® G HBcAb-N (Fujirebio, Japan) assay for IgG antibodies to hepatitis B core antigen (anti-HBc IgG) measurement in serum of subjects with overt and occult HBV infection (OBI). MATERIALS/METHODS:Serum anti-HBc IgG was assessed in 181 anti-HBc...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2020.114985
更新日期:2020-04-01 00:00:00
abstract::Four lateral flow assays (LFAs) were evaluated for the detection of purified botulinum neurotoxin A, toxin complex, and unpurified culture supernatant. They included the BioThreat (Tetracore, Rockville, MD), SMART (New Horizons Diagnostics, Columbia, MD), BADD (ADVNT Biotechnologies, Phoenix, AZ), and RAMP (Response B...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2006.07.017
更新日期:2007-03-01 00:00:00
abstract::Potential tigecycline-Enterobacteriaceae susceptibility breakpoints were evaluated using 2 approaches, which differed in the nature of the probabilities assessed by MIC value. Using a previously derived tigecycline population pharmacokinetic model and Monte Carlo simulation, a probability density function of steady-st...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2008.09.014
更新日期:2009-01-01 00:00:00
abstract::In a randomized, multicenter, open-label study, 490 ambulatory adult patients with lower respiratory tract infection (LRTI) were randomized to roxithromycin (ROX) 150 mg b.i.d. orally (n = 244) or amoxicillin plus clavulanic acid (AMX + CA) as 500 mg AMX + 125 mg CA t.i.d orally (n = 24). Clinical results were analyze...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 临床试验,杂志文章,多中心研究,随机对照试验
doi:10.1016/0732-8893(92)90133-e
更新日期:1992-05-01 00:00:00
abstract::Each clinical microbiology laboratory must establish its own standard battery of antimicrobial agents to be tested routinely on clinical isolates of various organism groups. Some choices are based upon the intrinsic activities of antimicrobial agents for a particular group of organisms, for example, agents primarily a...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/0732-8893(93)90117-p
更新日期:1993-03-01 00:00:00
abstract::Between February 2005 and January 2006 in Srinagarind Hospital, Thailand, 44 from 1730 isolates (2.5%) of Escherichia coli and 8 from 982 isolates (0.8%) of Klebsiella pneumoniae were found to produce plasmid-mediated AmpC β-lactamases (pAmpCs) as detected by a cefoxitin-Hodge test followed by a multiplex polymerase c...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2010.06.014
更新日期:2010-11-01 00:00:00
abstract::We analyzed sequences of graSR, vraSR, walKR and rpoB genes in hVISA from Brazil. Five isolates showed mutations in at least one gene. rpoB H481N and graS T224I were the most frequent mutations, followed by graR D148Q and walK A468T. Our study reinforces the heterogeneity of genetic patterns among hVISA. ...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2019.06.006
更新日期:2019-11-01 00:00:00
abstract::Combining a polymerase chain reaction (PCR) test with bronchoscopy is frequently performed to allow a rapid diagnosis of smear-negative pulmonary tuberculosis (PTB). However, limited data are available concerning clinical judgment in patients with suspected PTB and AFB smear- and PCR-negative bronchial aspirates (BA)....
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2012.03.019
更新日期:2012-07-01 00:00:00
abstract::Cycling Probe Technology (CPT) was combined with a colorimetric enzyme-immuno assay (EIA) to develop two assays for the detection of vanA and vanB genes in vancomycin resistant enterococci (VRE). The CPT-EIA assay employs a gene-specific fluorescein labeled DNA-RNA-DNA probe that gets cleaved within the probe : target...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/s0732-8893(99)00149-2
更新日期:2000-05-01 00:00:00
abstract::Molecular epidemiology allows us to know local HIV transmission and to design strategies of prevention. We studied 25 HIV newly diagnosed patients with the E138A mutation since the year 2010. Most transmission networks involved young and promiscuous men who have sex with men. Recent infection was only documented in pa...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2016.06.014
更新日期:2016-09-01 00:00:00
abstract::Over a five-year period (1995-1999) the Microbial Diseases Laboratory received 34 strains of E. coli O157:H7 each with a single aberrant biochemical property. In addition, 27 O157 strains with negative or delayed motility were noted during the same time period. These observations suggest that there may be an increased...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/s0732-8893(00)00196-6
更新日期:2000-11-01 00:00:00
abstract::The activities of trovafloxacin and ciprofloxacin against 38 strains of non-beta-lactamase-producing enterococci, resistant to ampicillin, 34 strains susceptible to ampicillin, and 3 vancomycin-resistant enterococci were studied. Trovafloxacin was more active than ciprofloxacin against all the enterococci studied. The...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/s0732-8893(97)00161-2
更新日期:1997-12-01 00:00:00
abstract::The impact of accessory gene regulator (agr) dysfunction and high bacterial density on vancomycin killing and resistance was evaluated among 10 clinical methicillin-resistant Staphylococcus aureus bloodstream isolates using time kill experiments. Under conditions of high inocula and agr dysfunction, vancomycin activit...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2009.01.028
更新日期:2009-06-01 00:00:00
abstract::As part of the SENTRY Antimicrobial Surveillance Program, 1562 bacterial isolates were recovered from hospitalized patients with skin and soft tissue infections (SSTIs) in 30 United States (U.S.) and 8 Canadian medical centers between October and December, 1997. The overall rank order of recovery of the six most commo...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章,多中心研究
doi:10.1016/s0732-8893(98)00162-x
更新日期:1999-05-01 00:00:00
abstract::Cellulomonas spp. are often believed to be of low virulence and have never been reported as a pathogen causing human disease before. We report the first case of endocarditis caused by Cellulomonas and complicated with osteomyelitis of the lumbar spine in a 78-year-old woman. General weakness and aggravated lower back ...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章,评审
doi:10.1016/j.diagmicrobio.2009.06.001
更新日期:2009-10-01 00:00:00
abstract::In febrile neutropenic patients, prompt empiric antimicrobial intervention is mandatory. Numerous studies have demonstrated the benefit of broad-spectrum beta-lactams active against Gram-negative aerobes as well as against streptococci and Staphylococcus aureus in this setting. With this interventional strategy, a red...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章,评审
doi:10.1016/s0732-8893(99)00024-3
更新日期:1999-07-01 00:00:00
abstract::Molecular (polymerase chain reaction [PCR]) methods are increasingly used to detect and type human enteroviruses (HEVs) and parechoviruses (HPeV). Here, we assessed their value in comparison to virus culture and serotyping for detection and typing of HEV and HPeV in stool samples from hospitalized patients. By use of ...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2010.05.016
更新日期:2010-10-01 00:00:00
abstract::Doripenem, a broad-spectrum parenteral carbapenem, has potency and pharmacokinetic/pharmacodynamic features most similar to imipenem and meropenem. Because of potential delays in release of commercial testing devices post-regulatory approval (US Food and Drug Administration), "surrogate markers" offer immediate suscep...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2007.10.018
更新日期:2007-12-01 00:00:00
abstract::The mechanism and clinical relevance of the inhibitory effect of ciprofloxacin on the metabolism of selected drugs were studied in patients with bacterial infections. In study A, antipyrine tests were carried out in two groups of patients taking 1000 mg (group 1) and 250 mg (group 2) of oral ciprofloxacin for 7-10 day...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/0732-8893(90)90096-e
更新日期:1990-03-01 00:00:00
abstract::We developed and evaluated a method to genotypically identify enterotoxigenic Escherichia coli (ETEC) and to characterize these organisms with respect to 18 of 21 known colonization factors (CFs). The method, which is based on polynucleotide DNA-DNA colony hybridization, includes a pooled toxin probe assay to identify...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/s0732-8893(02)00504-7
更新日期:2003-02-01 00:00:00
abstract::A shift to Candida non-albicans infections has been noted during the last years, and the emergence of multi-resistant Candida auris has complicated their management. The aim of this study was first to compare the performance of the novel chromogenic medium CHROMagar™ Candida Plus (CHROMagar, France) with CHROMagar™ Ca...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2020.115168
更新日期:2020-12-01 00:00:00
abstract::The Alere-i™ Influenza A&B (Abbott), a nicking endonuclease amplification reaction test, has recently been improved in order to deliver results in a few minutes. Our field observation highlights two problems with this new version: improper interpretation of a test as valid despite improper reagent hydration and falsel...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2019.03.002
更新日期:2019-08-01 00:00:00
abstract::The Verigene Gram-positive blood culture assay (BC-GP) is a microarray-based rapid diagnostic test, which includes targets for 12 bacterial species and 3 resistance determinants. We prospectively compared the diagnostic accuracy of the BC-GP to routine microbiologic methods and evaluated the potential of the BC-GP for...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2014.09.025
更新日期:2015-01-01 00:00:00
abstract:BACKGROUND:The Clostridium difficile rate in symptomatic patients represents both those with C. difficile infection (CDI) and those with colonization. To predict the extent of CDI overdiagnosis, we compared the asymptomatic colonization rate to the symptomatic positivity rate in hospitalized patients using nucleic acid...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2016.12.014
更新日期:2017-04-01 00:00:00
abstract::Bartonella quintana, known to cause various clinical symptoms, is increasingly recognized as one important cause of culture-negative endocarditis. We report a case of infectious endocarditis with B. quintana on the prosthetic valve, accompanied by proteinase 3-antineutrophil cytoplasmic antibody-positive collagen vasc...
journal_title:Diagnostic microbiology and infectious disease
pub_type: 杂志文章
doi:10.1016/j.diagmicrobio.2011.03.011
更新日期:2011-07-01 00:00:00