Rapid detection, identification, and enumeration of Pseudomonas aeruginosa in bottled water using peptide nucleic acid probes

被引:31
作者
Stender, H
Broomer, A
Oliveira, K
Perry-O'Keefe, H
Hyldig-Nielsen, JJ
Sage, A
Young, B
Coull, J
机构
[1] Boston Probes Inc, Bedford, MA 01730 USA
[2] Millipore Corp, Bedford, MA 01730 USA
关键词
chemiluminescent in situ hybridization; Pseudomonas aeruginosa; bottled water; detection; enumeration;
D O I
10.1016/S0167-7012(00)00196-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new chemiluminescent in situ hybridization (CISH) method that provides simultaneous detection, identification, and enumeration of Pseudomonas aeruginosa in bottled water within 1 working day has been developed. Individual microcolonies of P. aeruginosa were detected directly on membrane filters following 5 h of growth by use of soybean peroxidase-labeled peptide nucleic acid (PNA) probes targeted to a species-specific sequence in P. aeruginosa rRNA. Within each micro-colony, reaction of the peroxidase with a chemiluminescent substrate generated light that was subsequently captured by film or with a digital camera system. Each spot of light represented one micro-colony of P. aeruginosa. Sensitivity and specificity for the identification of P. aeruginosa were 100% as determined by testing 28 P. aeruginosa strains and 17 other bacterial species that included closely related Pseudomonas species. Furthermore, the number of micro-colonies of P. aeruginosa represented by light spots correlated with counts of visible colonies following sustained growth. We conclude that PNA CISH speeds up traditional membrane filtration techniques and adds the specificity of PNA probe technology to generate fast and definitive results. (C) 2000 Elsevier Science BN. All rights reserved.
引用
收藏
页码:245 / 253
页数:9
相关论文
共 33 条
  • [1] [Anonymous], STANDARD METHODS EXA
  • [2] COULL JM, 1999, Patent No. 0002908
  • [3] PHYLOGENETIC STAINS - RIBOSOMAL RNA-BASED PROBES FOR THE IDENTIFICATION OF SINGLE CELLS
    DELONG, EF
    WICKHAM, GS
    PACE, NR
    [J]. SCIENCE, 1989, 243 (4896) : 1360 - 1363
  • [4] EGHOLM M, 1993, NATURE, V365, P556
  • [5] European Union, 1980, OFFICIAL J EUROPEA L, VL229, P1
  • [6] PSEUDOMONAS-AERUGINOSA - GROWTH IN DISTILLED WATER FROM HOSPITALS
    FAVERO, MS
    CARSON, LA
    BOND, WW
    PETERSEN, NJ
    [J]. SCIENCE, 1971, 173 (3999) : 836 - &
  • [7] THE PHYLOGENY OF PROKARYOTES
    FOX, GE
    STACKEBRANDT, E
    HESPELL, RB
    GIBSON, J
    MANILOFF, J
    DYER, TA
    WOLFE, RS
    BALCH, WE
    TANNER, RS
    MAGRUM, LJ
    ZABLEN, LB
    BLAKEMORE, R
    GUPTA, R
    BONEN, L
    LEWIS, BJ
    STAHL, DA
    LUEHRSEN, KR
    CHEN, KN
    WOESE, CR
    [J]. SCIENCE, 1980, 209 (4455) : 457 - 463
  • [8] HOIBY N, 1982, ACTA PAEDIATR SC S, V301, P33
  • [9] HYLDIGNIELSEN JJ, 1999, PNA PROBES PROBE SET
  • [10] DEOXYRIBONUCLEIC-ACID SIMILARITIES AMONG PSEUDOMONAS SPECIES
    JOHNSON, JL
    PALLERONI, NJ
    [J]. INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY, 1989, 39 (03): : 230 - 235