Combination of ATP-bioluminescence and PNA probes allows rapid total counts and identification of specific microorganisms in mixed populations.
We have combined ATP-dependent bioluminescence with a novel chemiluminescent in situ hybridization (CISH) method using peroxidase-labeled peptide nucleic acid (PNA) probes targeting species-specific rRNA sequences to provide total counts and subsequent identification of specific microorganisms. Both methods are applied to the same membrane filter following a short incubation time and both methods provide results in the form of spots of light that are captured by the MicroStar detection system. Each spot of light represents individual micro-colonies detected by either ATP bioluminescence or PNA CISH. This new concept is particularly intended for in process and quality control of non-sterile products to rapidly provide total counts as well as presence/absence of specific indicators and/or pathogens in non-sterile, filterable samples.
Stender
H
H
Boston Probes, Inc., 75E Wiggins Avenue, 01730, Bedford, MA, USA. HStender@BostonProbes.com
Sage
A
A
Oliveira
K
K
Broomer
A J
AJ
Young
B
B
Coull
J
J
D000255:Adenosine Triphosphate / Q000378:metabolism*; D015169:Colony Count, Microbial; D015342:DNA Probes; D004926:Escherichia coli / Q000145:classification / Q000302:isolation & purification; D006090:Gram-Negative Bacteria / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D017403:In Situ Hybridization / Q000379:methods*; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D008851:Micropore Filters; D009696:Nucleic Acids / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D009195:Peroxidase; D011550:Pseudomonas aeruginosa / Q000145:classification / Q000302:isolation & purification; D012335:RNA, Ribosomal / Q000032:analysis; D012475:Salmonella / Q000145:classification / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013045:Species Specificity
D015342:DNA Probes; D007202:Indicators and Reagents; D009696:Nucleic Acids; D010455:Peptides; D012335:RNA, Ribosomal; D000255:Adenosine Triphosphate; D009195:Peroxidase