[Simple and efficient method for clinical isolation of Bordetella pertussis].
Commercially available Amies transport medium with charcoal and three isolation media were tested to assess their efficiency in the clinical isolation of B. pertussis. First, the isolation rates of B. pertussis were compared between direct inoculation of nasopharyngeal specimens and inoculation after stored in Amies transport medium for 8 hours or less. The comparative isolation rates were 81% both (35 of 43 specimens from 29 patients) for direct inoculation and transport medium. Second, nasopharyngeal specimens were incubated for 5 days at 35 degrees C on the three media; Bordet-Gengou Medium (BG) with 5 micrograms of CEX per ml, Cyclodextrin Solid Medium (CSM) with 5 micrograms of CEX per ml, and Charcoal Agar with 40 micrograms of CEX per ml. The organism was detected from 44 nasopharyngeal specimens from 20 patients on at least one of the three tested media. The comparative isolation rates were 91% (40 of 44) on BG with 5 micrograms of CEX per ml, 93% (41 of 44) on CSM with 5 micrograms of CEX per ml, and 91% (40 of 44) on CA with 40 micrograms of CEX per ml. Although no differences in the isolation rates were observed among the three media, the appearance of the colonies was earlier by one day on BG than the rest of the two media. The detection of B. pertussis was occasionally easier on CA than the rest of the two because of its higher suppression for nasopharyngeal flora. CSM has its advantage in that it does not need any blood and can be prepared at anytime. Also, the shelf life of these three media proved to be at least one month when stored at 10 degrees C. We conclude that the clinical isolation of B. pertussis was highly successful with the following simple procedure: nasopharyngeal specimens stored in Amies transport medium and inoculated on one of the three media, BG, CSM, or CA, and then incubated for 5 days at 35 degrees C.
Aoyama
T
T
Department of Pediatrics, School of Medicine, Keio University.
Goto
A
A
Iwai
H
H
Murase
Y
Y
Iwata
T
T
Takeuchi
Y
Y
Tonooka
H
H
D001431:Bacteriological Techniques; D001886:Bordetella pertussis / Q000302:isolation & purification*; D003470:Culture Media; D006801:Humans
D003470:Culture Media