Six hundred and ninety-nine strains of Neisseria gonorrhoeae were examined by the agar dilution method according to the M7-A5 guidelines established by the National Committee for Clinical Laboratory Standards (NCCLS) determine to their beta-lactamase production and susceptibility to penicillin G, cefixime, ceftriaxone, tetracycline, spectinomycin, ciprofloxacin, and azithromycin. The frequency of isolation of penicillinase-producing N. gonorrhoeae decreased gradually, from 7.3% of the test isolates (55 isolates) in 1995 to about 1% in 1998 and 1999. In contrast, while beta-lactamase-nonproducing N. gonorrhoeae exhibiting chromosomally mediated penicillin G resistance were not isolated from clinical specimens in 1995, the incidence of isolation of such resistant strains increased markedly, to 8.2% of 159 isolates, in 1997 and 14.9% of 242 isolates in 1999. The incidence of the isolation of tetracycline-resistant strains also increased between 1996 (none detected) and 1998-1999 (each 7%-8%), and a tendency towards an increase in the frequency of isolates of ciprofloxacin-resistant strains was also observed between 1995 (9.8%) and from 1997 to 1999 (more than 20%). There were no isolates resistant to any two antibiotics among penicillin G, tetracycline, and ciprofloxacin until 1997, but, in subsequent surveys in recent years, multidrug-resistant isolates (resistant to penicillin G, tetracycline, and ciprofloxacin) were detected in 1999.