Objective: To study Salmonella enterica serotype Typhi by pulsed-field gel electrophoresis in Hong Kong. Materials and Methods: One hundred thirty four isolates of Salmonella enterica serotype Typhi collected from 17 public hospitals and clinics in Hong Kong from 2000 to 2004 were studied in relation to epidemiological and clinical events. Isolates originated from 80 patients, with 29 patients providing multiple isolates. Susceptibility to six antibiotics was tested: ampicillin, trimethoprim-sulfamethoxazole, tetracycline, chloramphenicol, nalidixic acid, and ciprofloxacin. Strains were further subtyped by pulsed-field gel electrophoresis (PFGE) by separating XbaI-restricted genomic DNA of isolates. PFGE patterns that were shared between strains were further examined using restriction enzyme BlnI. Results: Of 134 S. Typhi isolates, 29 (21.6%) were resistant to at least one and up to five of the antibiotics tested. Using restriction fragments between 20 and 700 kb for analysis, the number of fragments generated by XbaI ranged from 14 to 20. Sixty-six distinct subtypes were identified in the first isolates of all 80 patients (epidemiologically unrelated) with a Simpson index of 0.993, indicating a high degree of diversity among these S. Typhi isolates. Multidrug-resistant and travel-associated S. Typhi appeared to cluster more closely than the rest of strains. Further analysis of PFGE patterns investigated the temporal relationships between the 83 strains collected from the 29 patients who gave multiple isolates. Conclusion: Dual infections or variants of the same isolates in the same patient occurred during the course of follow-up. These findings imply that PFGE data could be a valuable tool in predicting relapse, evaluating new antimicrobial drugs, and controlling the spread of typhoid disease. A regional, as well as global, typhoid bacillus fingerprint database should be set up to improve epidemiological investigations, as clinical cases easily move across national boundaries.

A total of 88 salmonella isolates (72 clinical isolates for which the ciprofloxacin MIC was >0.06 microg/ml, 15 isolates for which the ciprofloxacin MIC was < or =0.06 microg/ml, and Salmonella enterica serotype Typhimurium ATCC 13311) were studied for the presence of genetic alterations in four quinolone resistance genes, gyrA, gyrB, parC, and parE, by multiplex PCR amplimer conformation analysis. The genetic alterations were confirmed by direct nucleotide sequencing. A considerable number of strains had a mutation in parC, the first to be reported in salmonellae. Seven of the isolates sensitive to 0.06 micro g of ciprofloxacin per ml had a novel mutation at codon 57 of parC (Tyr57-->Ser) which was also found in 29 isolates for which ciprofloxacin MICs were >0.06 micro g/ml. Thirty-two isolates had a single gyrA mutation (Ser83-->Phe, Ser83-->Tyr, Asp87-->Asn, Asp87-->Tyr, or Asp87-->Gly), 34 had both a gyrA mutation and a parC mutation (29 isolates with a parC mutation of Tyr57-->Ser and 5 isolates with a parC mutation of Ser80-->Arg). Six isolates which were isolated recently (from 1998 to 2001) were resistant to 4 micro g of ciprofloxacin per ml. Two of these isolates had double gyrA mutations (Ser83-->Phe and Asp87-->Asn) and a parC mutation (Ser80-->Arg)(MICs, 8 to 32 microg/ml), and four of these isolates had double gyrA mutations (Ser83-->Phe and Asp87-->Gly), one parC mutation (Ser80-->Arg), and one parE mutation (Ser458-->Pro)(MICs, 16 to 64 micro g/ml). All six of these isolates and those with a Ser80-->Arg parC mutation were S. enterica serotype Typhimurium. One S. enterica serotype Typhi isolate harbored a single gyrA mutation (Ser83-->Phe), and an S. enterica serotype Paratyphi A isolate harbored a gyrA mutation (Ser83-->Tyr) and a parC mutation (Tyr57-->Ser); both of these isolates had decreased susceptibilities to the fluoroquinolones. The MICs of ciprofloxacin, levofloxacin, and sparfloxacin were in general the lowest of those of the six fluoroquinolones tested. Isolates with a single gyrA mutation were less resistant to fluoroquinolones than those with an additional parC mutation (Tyr57-->Ser or Ser80-->Arg), while those with double gyrA mutations were more resistant.

A total of 217 and 73 strains of Salmonella enterica serotype Typhi isolated from 1985 to 1997 in Hong Kong and in 2 months of 1989 and 1990 in Vietnam, respectively, were studied. These isolates were typed by plasmid profile analysis, plasmid fingerprinting, ribotyping with PstI, and total DNA fingerprinting with NarI. There appeared to be no major outbreak of typhoid fever in Hong Kong during the study period since there was considerable heterogeneity among the isolates. Isolates from Hong Kong were different from those from Vietnam. Thirty-seven percent of Vietnamese isolates belonged to two predominant clones, with the rest being heterogeneous in nature. Total DNA fingerprinting supplemented with ribotyping could be a reliable and rapid method for epidemiological typing of S. enterica serotype Typhi.