Mei-Feng Lee

Chi-Mei Medical Center, 臺南市, Taiwan, Taiwan

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Publications (18)38.35 Total impact

  • Mei-Feng Lee, Jing-Yu Chen, Chien-Fang Peng
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    ABSTRACT: In this study, we designed a novel colorimetric method to detect multidrug resistance in Mycobacterium tuberculosis isolates. The assay of loop-mediated isothermal amplification (LAMP) is used to amplify target DNA from multidrug-resistant M. tuberculosis isolates, and enzyme-linked immunosorbent assay (ELISA) is used for the colorimetric determination. This method is designed based on point mutation at the hot spot region in target drug-resistant gene using LAMP-polymerase chain reaction (PCR), hybridization, and thermal melting for differentiating homoduplex DNA (drug-susceptible stain) and heteroduplex DNA (resistance mutant). From ELISA colorimetric detection, color change developed in drug-susceptible strains, and colorless result appeared in resistance mutants. A comparison of this LAMP-PCR-hybridization–thermal melt–ELISA (LAMP–TM–ELISA) method with the automated BACTEC MGIT 960 system showed that the sensitivity of this molecular analysis of resistance to isoniazid, rifampin, amikacin, and ciprofloxacin in M. tuberculosis was 92.3%, 95.3%, 93.1%, and 91.4%, respectively. This method for detection of resistance to isoniazid, rifampin, amikacin, and ciprofloxacin in M. tuberculosis showed a specificity of 95.5–98.2% and a test efficiency of 93.2–96.8%. This LAMP–TM–ELISA method will be a useful tool for rapid diagnosis (within 1 working day) and cost-effectiveness (US$15/reaction) to detect resistance to isoniazid, rifampin, amikacin, and ciprofloxacin via katG, inhA and mabA-inhA promoter, rpoB, rrs, gyrA, and gyrB genes in M. tuberculosis isolates.
    Biomarkers and Genomic Medicine. 09/2014;
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    ABSTRACT: Mutations in the quinolone resistance-determining regions (QRDRs) of gyrA, gyrB, parC, and parE have been characterized among the 232 isolates of ciprofloxacin (CIP)-resistant Pseudomonas aeruginosa. As expected, no mutations in the QRDRs of four target genes were detected in the CIP-susceptible isolates of P. aeruginosa. It was noted that P. aeruginosa showing no mutation in the QRDRs of target genes were frequently found in isolates with a CIP in minimal inhibitory concentration (MIC) = 2 μg/mL than those of isolates with a CIP in MIC ≥4 μg/mL. The prevalence of P. aeruginosa with no mutations in the QRDRs of target genes is higher in isolates only resistant to CIP than in isolates resistant to CIP and other drugs. Double mutations occurring in gyrA and parC genes associated with a high-level resistance to CIP in MICs ≥4 μg/mL were found in 101 out of 176 isolates. Furthermore, mutations in parC and parE joined with mutation in gyrA were commonly found in P. aeruginosa highly resistant to CIP.
    Biomarkers and Genomic Medicine. 06/2014; 6(2):79–83.
  • Ke-Yu Hsiao, Mei-Feng Lee, Chien-Fang Peng
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    ABSTRACT: In this study, detection and characterization of class 1 integron-associated gene cassettes from Pseudomonas aeruginosa isolates in southern Taiwan were investigated. The study focused on the association between integron-associated resistance gene cassettes and multidrug resistance in P. aeruginosa isolates. Using polymerase chain reaction amplification, DNA sequencing, and basic local alignment search tool analysis, a total of 22 different types of gene cassette arrays were detected in 162 class 1 integron-positive P. aeruginosa isolates. We first identified 11 different types of new gene cassette arrays within class 1 integron in P. aeruginosa isolates, including aac(6′)-II-catB2-aadA2, aac(6′)-II-aadA2, aac(6′)-II-catB2, aacA4-aadA15, aacC1-orfA-orfB-aadA1, cm1A-aadA1, catB3-blaOxA-10-aadA15, aacA4-catB8-aadA1, aadB-orfF1-aadA11, dfrB1, and dfrB4a-aacA4-aacA4-aadA1. Of these, aac(6′)-II-catB2-aadA2 was the most frequently found gene cassette. Twenty-one (21/162, 12.9%) strains carrying two different types of gene cassette arrays of catB3-blaOxA-10-aadA15 and aac(6′)-II-catB2-aadA2 were also simultaneously present in the P. aeruginosa isolates. A novel dfrB4a gene, different from the dfrB4 gene detected in the gene cassette array of dfrB4a-aacA4-aacA4-aadA, was characterized. A metallo-β-lactamaseo gene was also found to be carried on the gene cassette of blaVIM-3-orf2a-aacA4-aadB-aacA4 inserted in a class 1 integron obtained from meropenem-resistant P. aeruginosa isolates.
    Biomarkers and Genomic Medicine. 06/2014; 6(2):74–78.
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    ABSTRACT: This study was conducted to investigate an outbreak caused by imipenem-resistant Acinetobacter baumannii (IRAB) in a medical intensive care unit (ICU) in a regional hospital.
    PLoS ONE 01/2014; 9(9):e107975. · 3.53 Impact Factor
  • The American journal of emergency medicine 10/2013; · 1.54 Impact Factor
  • Journal of the Formosan Medical Association 09/2013; · 1.00 Impact Factor
  • The Journal of infection 04/2013; · 4.13 Impact Factor
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    ABSTRACT: The widespread multidrug-resistant Enterobacteriaceae pose a serious therapeutic challenge. Colistin and tigecycline are potential antimicrobial agents for treating infections caused by extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae. We evaluated the in-vitro activity of colistin sulfate against 253 ESBL producers isolated from patients admitted to a medical center in southern Taiwan (Escherichia coli, n = 82; Klebsiella pneumoniae, n = 102; Enterobacter cloacae, n = 34; and Serratia marcescens, n = 35). Colistin showed promising in-vitro activity against E. coli, K. pneumoniae, and E. cloacae, but not S. marcescens. One ESBL-producing K. pneumoniae strain with resistance to carbapenems (ertapenem, imipenem, and meropenem) was selected for time-killing studies. A combination of colistin and tigecycline showed synergism, but there was an inoculum effect. In conclusion, colistin was active against most ESBL-producing Enterobacteriaceae, and a combination of colistin with tigecycline was synergistic against some highly resistant strains, even those with carbapenem resistance.
    Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi 01/2013; · 1.63 Impact Factor
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    ABSTRACT: Infections due to Prototheca spp. are ubiquitous in nature, occurring in both immunocompetent and immunocompromised patients. The study cohort consisted of 14 cases of Prototheca algaemia reported over the past 5 decades and 2 recent cases from study hospitals. Prototheca wickerhamii was the most common species. The overall mortality rate was 62.5%. Prototheca algaemia, a healthcare-associated infection, was observed in immunocompromised patients and was associated with a poor prognosis.
    Japanese journal of infectious diseases. 01/2013; 66(6):523-525.
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    ABSTRACT: Although extended-spectrum-β-lactamase (ESBL)-producing aeromonads have been increasingly reported in recent years, most of them were isolates from case reports or environmental isolates. To investigate the prevalence of ESBL producers among Aeromonas blood isolates and the genes encoding ESBLs, consecutive nonduplicate Aeromonas blood isolates collected at a medical center in southern Taiwan from March 2004 to December 2008 were studied. The ESBL phenotypes were examined by clavulanate combination disk test and the cefepime-clavulanate ESBL Etest. The presence of ESBL-encoding genes, including bla(TEM), bla(PER), bla(CTX-M), and bla(SHV) genes, was evaluated by PCR and sequence analysis. The results showed that 4 (2.6%) of 156 Aeromonas blood isolates, 1 Aeromonas hydrophila isolate and 3 Aeromonas caviae isolates, expressed an ESBL-producing phenotype. The ESBL gene in two A. caviae isolates was bla(PER-3), which was located in both chromosomes and plasmids, as demonstrated by Southern hybridization. Of four patients with ESBL-producing Aeromonas bacteremia, two presented with catheter-related phlebitis and the other two with primary bacteremia. Three patients had been treated with initial noncarbapenem β-lactams for 5 to 10 days, and all survived. In conclusion, ESBL producers exist among Aeromonas blood isolates, and clinical suspicion of ESBL production should be raised in treating infections due to cefotaxime-resistant Aeromonas isolates.
    Antimicrobial Agents and Chemotherapy 10/2011; 55(12):5813-8. · 4.57 Impact Factor
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    ABSTRACT: By using inverse PCR and DNA sequencing, 13 sul3-associated mutational integrons, 2 defective class 1 integrons, and 1 qnrB2-associated complex sul1-type class 1 integrons were identified in Salmonella enterica serovar Choleraesuis, Pseudomonas aeruginosa, and Enterobacter cloacae, respectively. In addition, conjugation and Southern hybridization demonstrated that unusual class 1 integrons were located on plasmids or integrated into chromosomal DNA. Thus, an inverse PCR assay can be a valuable tool for the analysis of unusual structures of the 3' conserved region of class 1 integrons.
    Antimicrobial Agents and Chemotherapy 02/2011; 55(2):943-5. · 4.57 Impact Factor
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    ABSTRACT: In this study, we designed a simple and rapid colorimetric detection method, a one-tube loop-mediated isothermal amplification (LAMP)-PCR-hybridization-restriction endonuclease-ELISA [one-tube LAMP-PCR-HY-RE-ELISA] system, to detect resistance to isoniazid, ethambutol and streptomycin in strains of Mycobacterium tuberculosis isolated from clinical specimens. The clinical performance of this method for detecting isoniazid-resistant, ethambutol-resistant and streptomycin-resistant isolates of M. tuberculosis showed 98.9%, 94.3% and 93.8%, respectively. This assay is rapid and convenient that can be performed within one working day. One-tube LAMP-PCR-HY-RE-ELISA system was designed based on hot spot point mutations in target drug-resistant genes, using LAMP-PCR, hybridization, digestion with restriction endonuclease and colorimetric method of ELISA. In this study, LAMP assay was used to amplify DNA from drug-resistant M. tuberculosis, and ELISA was used for colorimetrical determination. This assay will be a useful tool for rapid diagnosis of mutant codons in strains of M. tuberculosis for isoniazid at katG 315 and katG 463, ethambutol at embB 306 and embB 497, and streptomycin at rpsL 43.
    Journal of microbiological methods 10/2010; 83(1):53-8. · 2.43 Impact Factor
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    ABSTRACT: The specificity and regulation of GSK3beta are thought to involve in the docking interactions at core kinase domain because of the particular amino acid residues. Recent X-ray diffraction studies illuminated the relative binding residues on AxinGID and FRATtide for GSK3beta docking and appeared that GSK3beta Val267Gly (V267G) and Tyr288Phe (Y288F) could distinguish the direct interaction between AxinGID and FRATtide. In order to explore the mode that involved the binding of GSKIP to GSK3beta and compare it with that of AxinGID and FRATtide, we pinpointed the binding sites of GSKIP to GSK3beta through the single-point mutation of four corresponding sites within GSK3beta (residues 260-300) as scaffold-binding region I (designated SBR-I(260-300)). Our data showed that these three binding proteins shared similar binding sites on GSK3beta. We also found that the binding of GSK3beta V267G mutant to GSKIP and AxinGID, but not that of Y288F mutant (effect on FRATtide), was affected. Further, based on the simulation data, the electron-density map of GSKIPtide bore closer similarity to the map AxinGID than to that of FRATtide. Interestingly, many C-terminal helix region point-mutants of GSK3beta L359P, F362A, E366K, and L367P were able to eliminate the binding with FRATtide, but not AxinGID or GSKIP. In addition, CABYR exhibited a unique mode in binding to C-terminal helix region of GSK3beta. Taken together, our data revealed that in addition to the core kinase domain, SBR-I(260-300), another novel C-terminus helix region, designated SBR-II(339-383), also appeared to participate in the recognition and specificity of GSK3beta in binding to other specific proteins.
    Molecular and Cellular Biochemistry 06/2010; 339(1-2):23-33. · 2.33 Impact Factor
  • Mei-Feng Lee, Yen-Hsu Chen, Chien-Fang Peng
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    ABSTRACT: Integron-mediated multidrug resistance in Salmonellaenterica serovar Choleraesuis poses a major concern in human salmonellosis in Taiwan. In this study, 71 strains of S. Choleraesuis harbouring a class 1 integron from humans and animals were characterised for detection of integrons, gene cassettes and the spvC virulence gene using molecular genetic techniques. All 71 strains tested were negative for class 2 and 3 integrons. Fifty-eight (81.7%) of the isolates harboured a class 1 integron with different gene cassettes, including dfr12-orfF-aadA2 in 50 strains, dfrA1-UN in 7 strains and aadA1 in 1 strain. Among the 71 isolates, 1 isolate of S. Choleraesuis H40 harboured two class 1 integrons, the first integron with the dfr12-orfF-aadA2 gene cassette and the second integron with a novel single gene cassette aadA22, a new variant of the aadA gene family. Moreover, 13 of the 71 isolates were negatively amplified by 5'-CS and 3'-CS specific primer pairs and represented a novel gene cassette array of sat-psp-aadA2-cmlA1-aadA1-like-qacH-tnp-sul3 in class 1 integrons. In mating experiments, the transferability of integron-mediated drug resistance in S. Choleraesuis isolates was demonstrated. The class 1 integron-borne gene cassette dfr12-orfF-aadA2 was characterised as becoming predominant in multidrug-resistant S. Choleraesuis isolates. Furthermore, both the dfrA1-UN gene cassette and the sul3-associated novel gene cassette were also identified in the S. Choleraesuis isolates. To date, this is the first report describing the dissemination of a sul3-associated integron in S. Choleraesuis isolates in Taiwan.
    International journal of antimicrobial agents 01/2009; 33(3):216-22. · 3.03 Impact Factor
  • Mei-Feng Lee, Yen-Hsu Chen, Chien-Fang Peng
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    ABSTRACT: Traditional culture, followed by a panel of biochemical tests for the diagnosis of tuberculosis (TB), is time-consuming, and rapid identification of Mycobacterium tuberculosis is crucial for the early administration of appropriate therapy. In this study, the reverse transcription loop-mediated isothermal amplification combined with enzyme-linked immunosorbent hybridization (RT-LAMP-ELISA-hybridization) assay has been designed for the rapid detection of 16S rRNA in clinical isolates of M. tuberculosis. This assay reproducibly detected a single copy, as opposed to 2000 copies of MTB 16S rRNA detected by conventional gel electrophoresis. Among the 150 specimens of sputum analysed, RT-LAMP-ELISA-hybridization assay had a sensitivity of 94.1% in the culture method, compared to the Amplified M. tuberculosis Direct Test (AMTD), 91.1% and the 88.2% sensitivity of acid-fast staining. Furthermore, RT-LAMP-ELISA-hybridization assay is more cost-effective when compared to the real-time TaqMan RT-PCR and AMTD assays. In conclusion, our results suggest that the RT-LAMP-ELISA-hybridization assay is a highly sensitive, low cost diagnostic tool useful for the rapid and accurate direct diagnosis of sputum specimens, and is suitable for routine clinical use.
    Journal of Microbiological Methods 11/2008; 76(2):174-80. · 2.16 Impact Factor
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    ABSTRACT: This work studies antimicrobial resistance and class 1 integrons of Aeromonas spp. in human isolates from southern Taiwan. PCR amplification and DNA sequence analyses were performed to characterize the gene cassette regions of the class 1 integron in 204 isolates of Aeromonas hydrophila, 36 isolates of A. sobria, 23 isolates of A. veronii, and 4 isolates of A. caviae. By using Southern hybridization with an intI1 probe to determine the presence of class 1 integrons in the 9 isolates of Aeromonas spp. harboring plasmid DNA, only 2 isolates, one A. veronii AV69 harboring 176-kb plasmid DNA, and one A. hydrophila AH207 harboring 149-kb plasmid DNA were identified. A conjugation experiment was carried out with 2 isolates of A. veronii AV69 and A. hydrophila AH207. Only one transconjugant of Escherichia coli AH207, containing 149-kb plasmids obtained from A. hydrophila AH207, was identified. ERIC-PCR analysis was performed to analyze the genetic relatedness in all isolates of Aeromonas spp. that carry class 1 integrons. The results of cluster analysis in this experiment revealed that none of these isolates were clonal, which may indicate that they were not related to the outbreak. Among the 267 isolates tested, class 1 integrons were detected in 37 isolates (13.9%) of Aeromonas spp. from humans. No class 2 or class 3 integrons were detected in this study. Gene cassette structures were identified in 30 (81.0%) of 37 isolates of Aeromonas spp. containing class 1 integrons. The gene cassette of dfrA12-orfF-aadA2 was the most prevalent in the gene cassette array (16.0%), followed by arr3-aacA4 (13.3%) and dfr2d-catB3-aadA1 (10.0%). Four novel arrays of gene cassettes were also identified, namely, dfr2d-catB3-aadA1, aadA1-aac(6')-II, aadA4a, and aac(6')-II-blaOXA-21-catB3. This is the first report of Aeromonas spp. isolates from humans.
    Japanese journal of infectious diseases 10/2008; 61(5):343-9. · 1.51 Impact Factor
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    ABSTRACT: In this study, 260 non-replicate imipenem-resistant Gram-negative bacteria isolated between January 2002 and December 2006 were subjected to a screening test for detection of metallo-beta-lactamase (MBL) using the Etest containing imipenem and ethylene diamine tetra-acetic acid (EDTA). Among the 260 strains, 123 (47.3%) appeared to produce MBL. Of these 123 strains, 113 (91.9%) were found by polymerase chain reaction (PCR) to carry MBL genes of types blaVIM-2, blaVIM-3, blaVIM-11 (blaVIM-11a), blaIMP-8 and novel blaIMP-24. One strain of Serratia marcescens harboured two MBL genes (blaVIM-11 and blaIMP-8) simultaneously. Of the 123 strains, 116 strains (94.3%) carrying the intI1 gene and 21 strains carrying integron-associated blaVIM-3, blaVIM-11 and blaIMP-8 genes were identified among Acinetobacter baumannii, Pseudomonas aeruginosa, Acinetobacter haemolyticus and S. marcescens. Using pulsed-field gel electrophoresis (PFGE) and Southern hybridisation with the blaVIM gene probe for I-CeuI-digested genomic DNA, P. aeruginosa 9527 strain harboured two class 1 integron-associated MBL genes in the chromosome, including blaVIM-3-orf2-aacA4 and novel bla(VIM-3)-orf2-aacA4-aadB-aacA4. This is the first description of the blaVIM-11 gene spreading among P. aeruginosa and A. baumannii strains in southern Taiwan. This finding suggests that clinical spread of this blaVIM-11 gene is a matter of great concern for carbapenem resistance in southern Taiwan.
    International Journal of Antimicrobial Agents 10/2008; 32(6):475-80. · 4.42 Impact Factor
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    ABSTRACT: The objective of this study was to characterize the gene cassettes of class 1 integrons and antimicrobial resistance among CTX-M-3-producing Serratia marcescens isolates from different specimens in southern Taiwan. One hundred and twenty-two isolates (70.5%) of 173 CTX-M-3-producing S. marcescens isolates were positive for class 1 integrons, including 53.3% of blood isolates, 94.1% of urine isolates, and 87.2% of sputum isolates. No class 2 or class 3 integrons were detected in this study. By PCR with primers 5'-CS and 3'-CS for the amplification of gene cassettes regions, amplicons ranging from 0.7 to 3.0 kb in length were found in 108 (88.5%) of the 122 class 1 integron-containing isolates of CTX-M-3-producing S. marcescens isolates. Ten different types by pattern of amplicons for class 1 integrons were obtained. The Type I amplicon (46.3%) harbors two different class 1 integrons containing the gene cassette of aadA2 and aadB-catB3, respectively, and was most prevalent in the gene cassette region-positive S. marcescens isolates, followed by the Type II amplicon, which harbors one class 1 integron containing the gene cassette dfrA12-orfF-aadA2 (28.7%). Most of the S. marcescens isolates (66.7%, 8/12) harboring three different class 1 integrons (Type IV amplicon) were found in blood isolates. Class 1 integrons were conjugally transferred to recipients in 92.0% of S. marcescens harboring two different class 1 integrons containing the gene cassettes aadA2 and aadB-catB3, respectively. The transfer rate of class 1 integron carrying dfrA12-orfF-aadA2 was detected in 77.4% of S. marcescens isolates. The results showed that all those isolates with conjugative transfer of integrons carried their class 1 integrons on the conjugative plasmids.
    Japanese journal of infectious diseases 10/2007; 60(5):250-6. · 1.51 Impact Factor

Publication Stats

84 Citations
38.35 Total Impact Points

Institutions

  • 2011–2014
    • Chi-Mei Medical Center
      臺南市, Taiwan, Taiwan
  • 2008–2014
    • Kaohsiung Medical University
      • • Department of Medical Laboratory Science and Biotechnology
      • • College of Medicine
      Kao-hsiung-shih, Kaohsiung, Taiwan
  • 2013
    • Chia Nan University of Pharmacy and Science
      臺南市, Taiwan, Taiwan