Real-time PCR and NASBA for rapid and sensitive detection of Vibrio cholerae in ballast water.
Transport of ballast water is one major factor in the transmission of aquatic organisms, including pathogenic bacteria. The IMO-guidelines of the Convention for the Control and Management of Ships' Ballast Water and Sediments, states that ships are to discharge <1 CFU per 100 ml ballast water of toxigenic Vibrio cholerae, emphasizing the need to establish test methods. To our knowledge, there are no methods sensitive and rapid enough available for cholera surveillance of ballast water. In this study real-time PCR and NASBA methods have been evaluated to specifically detect 1 CFU/100ml of V. cholerae in ballast water. Ballast water samples spiked with V. cholerae cells were filtered and enriched in alkaline peptone water before PCR or NASBA detection. The entire method, including sample preparation and analysis was performed within 7 h, and has the potential to be used for analysis of ballast water for inspection and enforcement control.
Norwegian Defence Research Establishment (FFI), P.O. Box 25, N-2027 Kjeller, Norway. else-marie.fykse@ffi.no
Else M
EM
Fykse
Trine
T
Nilsen
Agnete Dessen
AD
Nielsen
Ingun
I
Tryland
Stephanie
S
Delacroix
Janet M
JM
Blatny
D004784:Environmental Monitoring / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction; D012623:Seawater / Q000382:microbiology; D021161:Self-Sustained Sequence Replication; D012680:Sensitivity and Specificity; D012767:Ships; D013997:Time Factors; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D014871:Water Microbiology