An integrated culture and real-time PCR method to assess viability of disinfectant treated Bacillus spores using robotics and the MPN quantification method.
Using robotics and the MPN technique, a 96-microwell method was developed to compare two procedures for enumeration of viable chlorine-treated B. atrophaeus spores: broth-culture enrichment followed by real-time polymerase chain reaction analysis; and filter plating on agar. Recoveries of chlorine-treated spores were improved by broth enrichment over filter plating, whereas recoveries of non-treated spores were not different in the two procedures.
Varughese
Eunice A
EA
National Exposure Research Laboratory, U.S. Environmental Protection Agency, 26 W. Martin Luther King, Cincinnati, OH 45268, United States. varughese.eunice@epa.gov
Wymer
Larry J
LJ
Haugland
Richard A
RA
D001407:Bacillus / Q000254:growth & development / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D002452:Cell Count / Q000379:methods*; D003470:Culture Media; D004269:DNA, Bacterial; D004202:Disinfectants; D004203:Disinfection / Q000379:methods*; D008828:Microbiological Techniques; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012371:Robotics; D013171:Spores, Bacterial