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Biomedical subjects

Kai Man Kam

Publications and source records attributed to Kai Man Kam.

15 recordsLinked to original sources
PubMed2006

Rapid pulsed-field gel electrophoresis protocol for subtyping of Streptococcus suis serotype 2.

A rapid pulsed-field gel electrophoresis (PFGE) protocol for subtyping of Streptococcus suis serotype 2 was developed and evaluated using 27 clinical isolates from 22 epidemiologically unrelated patients. Results were matched against antibiogram, virulence genotyping and multi locus sequence typing (MLST). PFGE appeared to be the most discriminatory with numerical index of discrimination (D) equal to 0.87.

Animals↗
PubMed2006

Stepwise decrease in moxifloxacin susceptibility amongst clinical isolates of multidrug-resistant Mycobacterium tuberculosis: correlation with ofloxacin susceptibility.

To study the effectiveness of moxifloxacin (MOX) against multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB), in vitro drug susceptibility tests on MOX and ofloxacin (OFX) using the MGIT system against 132 nonduplicate MDR MTB isolates (108 OFX-sensitive and 24 OFX-resistant) were performed. Eleven OFX-resistant non-MDR MTB isolates were also included. All strains that were susceptible to OFX were shown to be also susceptible to MOX. For OFX-resistant isolates, regardless of their MDR status, a 4- to 8-fold decrease in MIC was observed for MOX when compared to OFX. On the basis of similarities in mode of action and pharmacokinetic data between MOX and OFX, a breakpoint MIC of 2 mg/L was suggested for MOX in the MGIT system. All but two OFX-resistant isolates possessed MIC of MOX of 2 mg/L or lower and were considered to be susceptible to MOX; however, all OFX-resistant strains studied had mutations involved in resistance to quinolones. DNA sequence analysis for the gyrase A mutation, which was an indicator for OFX resistance, correlated with MOX susceptibility changes. Significant decreases in MOX susceptibility amongst MDR-MTB strains were observed that correlated with OFX susceptibility. The Asp94Gly mutation appeared to be associated with a higher level of MICs to both OFX and MOX. These findings on the stepwise decline of in vitro susceptibility to MOX suggest that clinical usage of MOX should be accompanied by careful monitoring of susceptibility to this important anti-MDR MTB drug.

Anti-Bacterial Agents↗
PubMed2006

Optimization of variable number tandem repeat typing set for differentiating Mycobacterium tuberculosis strains in the Beijing family.

Variable number tandem repeat (VNTR) typing of Mycobacterium tuberculosis, as presently used, has often failed to differentiate clonal strains especially in areas where the Beijing family genotype is predominant. We have evaluated mycobacterial interspersed repetitive units (MIRUs), Queen's University of Belfast (QUB) loci and exact tandem repeat (ETR) loci individually for their abilities to differentiate the Beijing and non-Beijing genotype families of M. tuberculosis. The best locus was QUB 3232 followed by QUB 11b. Available MIRU, QUB and ETR loci were then consecutively arranged in declining order of discriminatory power, and combined to form an optimal set of 12-loci VNTR typing (MIRU 10, 26, 39, 40; QUB 11a, 11b, 15, 18, 26, 3232, 3336; ETR A) that possessed comparable discriminatory ability to IS6110 restriction fragment length polymorphism. This optimized 12-loci VNTR typing set can become an important tool for tracking M. tuberculosis of the Beijing family.

Bacterial Typing Techniques↗
PubMed2005

Plasmid-mediated resistance to ciprofloxacin and cefotaxime in clinical isolates of Salmonella enterica serotype Enteritidis in Hong Kong.

OBJECTIVES: To characterize the genetic determinants involved in the reduced susceptibility to ciprofloxacin and cefotaxime in Salmonella enterica serotype Enteritidis clinical isolates obtained from four patients in summer 2003 in Hong Kong. METHODS: Three Salmonella Enteritidis isolates from blood culture and one from stool were collected due to their increased resistance to ciprofloxacin and cefotaxime. PFGE analysis was used to investigate their genetic relatedness. Conjugation experiments were employed to show if the genetic determinants involved were plasmid-mediated. MICs of various antimicrobials were determined by VITEK 2 and Etest. Based on the susceptibility and conjugation experiment results, previously described PCR methods were employed to detect sequences homologous to qnr and bla(CTX-M) suspected to be involved in the reduced susceptibility to ciprofloxacin and cefotaxime, respectively. RESULTS: PFGE analysis showed that the four Salmonella isolates were clonally unrelated. The presence of a qnr-like gene and the CTX-M allele bla(CTX-M-14) on four different transferable plasmids harboured by the four isolates was confirmed. CONCLUSIONS: This is the first report of transferable fluoroquinolone resistance due to a new qnr allele, which appeared to be linked to bla(CTX-M-14), in isolates of Salmonella Enteritidis in Hong Kong.

Amino Acid Sequence↗
PubMed2005

Utility of mycobacterial interspersed repetitive unit typing for differentiating multidrug-resistant Mycobacterium tuberculosis isolates of the Beijing family.

Mycobacterial interspersed repetitive unit (MIRU) typing has been found to allow rapid, reliable, high-throughput genotyping of Mycobacterium tuberculosis and may represent a feasible approach to study global M. tuberculosis molecular epidemiology. To evaluate the use of MIRU typing in discriminating drug-resistant M. tuberculosis strains of the Beijing genotype family, 102 multidrug-resistant (MDR) clinical isolates and 253 randomly selected non-MDR isolates collected from 2000 to 2003 in Hong Kong were subjected to 12-locus MIRU typing, spoligotyping, and IS6110 restriction fragment length polymorphism (RFLP) typing. Spoligotyping showed that 243 (68.5%) of 355 isolates belonged to Beijing family genotype. MIRU typing showed lower discrimination in differentiating between the Beijing family strains (Hunter-Gaston discriminative index [HGI] of 0.8827) compared with the IS6110 RFLP method (HGI = 0.9979). For non-Beijing strains, MIRU typing provided discrimination (HGI = 0.9929) comparable to that of the RFLP method (HGI = 0.9961). There was no remarkable difference in discrimination power between the two methods in differentiating both within and between MDR and non-MDR strains of M. tuberculosis. Dendrograms constructed with the MIRU typing data showed a clear segregation between the Beijing and non-Beijing genotype. Addition of RFLP to MIRU typing offered a higher discrimination ability (92.6%) than did addition of MIRU typing to RFLP (40.0%). This supported the potential use of this method to analyze the global genetic diversity of MDR M. tuberculosis strains that may be at different levels of evolutionary divergence.

Bacterial Typing Techniques↗
PubMed2005

Use of variable-number tandem-repeat typing to differentiate Mycobacterium tuberculosis Beijing family isolates from Hong Kong and comparison with IS6110 restriction fragment length polymorphism typing and spoligotyping.

The Mycobacterium tuberculosis Beijing family isolates may cause more than a quarter of all tuberculosis cases worldwide, are emerging in some areas, and are often associated with drug resistance. Early recognition of transmission of this genotype is therefore important. To evaluate the usefulness of variable-number tandem-repeat (VNTR) typing to discriminate and recognize strains of the Beijing family, M. tuberculosis isolates from Hong Kong were subjected to VNTR analysis, spoligotyping, and IS6110 restriction fragment length polymorphism (RFLP) typing. The allelic diversity of the 14 VNTR loci included in the analysis varied from 0 to 0.618 among Beijing strains. The discriminatory power of VNTR analysis was slightly lower than that of IS6110 RFLP. Our analysis shows that VNTR typing, which has many practical advantages over RFLP typing, can be used for epidemiological studies of Beijing strains. However, VNTR-defined clusters should be subtyped with IS6110 RFLP for maximal resolution.

Bacterial Typing Techniques↗
PubMed2003

Routine use of a simple low-cost genotypic assay for the identification of mycobacteria in a high throughput laboratory.

A novel polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene was used for the routine identification of mycobacteria in a high throughput clinical laboratory. A total of 2036 clinical isolates were tested by PRA in conjunction with other methods. The PRA identification of M. tuberculosis complex was 100% sensitive and specific, and 74.5% of nontuberculous mycobacteria (NTM) were correctly identified. It gave highly consistent results for Mycobacterium avium complex (MAC) species and for most isolates of M. fortuitum, M. chelonae, and M. kansasii. It had proven to be highly robust and stable despite usage on such a large-scale and is thus particularly suitable for use in high throughput laboratories in areas with a high incidence of tuberculosis.

Algorithms↗
PubMed2003

Molecular characterization of quinolone-resistant Neisseria gonorrhoeae in Hong Kong.

In Hong Kong, ParC changes among high-level quinolone-resistant Neisseria gonorrhoeae (QRNG) isolates at Ser-87-->Arg were associated with a higher level of resistance than a Ser-87-->Ile alteration. Two previously undescribed mutations in clinical isolates occurring in gyrA, conferring Ala-92-->Pro and Asp-95-->Tyr changes, were detected. Nine different outer membrane lipoprotein (Lip) repeat classes-11 to 19 repeats-were identified, with repeat lengths of 16 and 17 the most common, indicating considerable strain diversity.

Amino Acid Sequence↗
PubMed2003

Molecular subtyping of Vibrio cholerae O1 and O139 by pulsed-field gel electrophoresis in Hong Kong: correlation with epidemiological events from 1994 to 2002.

Two hundred twenty isolates of Vibrio cholerae O1 and O139 collected from 1994 to 2002 in Hong Kong were analyzed by pulsed-field gel electrophoresis (PFGE). Chromosomal DNAs from all V. cholerae isolates in agarose plugs were digested with the restriction enzyme NotI, resulting in 20 to 27 bands. Sixty distinctive PFGE patterns in the range of 10 to 300 kb were noted among 213 isolates typeable by PFGE. By comparing the common PFGE patterns obtained from four well-defined outbreaks of V. cholerae O1 and O139 with those obtained from other, epidemiologically unrelated isolates during the study period, indistinguishable and similar PFGE patterns were identified, indicating their close relatedness, in agreement with the results of epidemiological investigations. Heterogeneous PFGE patterns (with four to six banding differences), however, were identified among strains that were imported from other parts of Asia, including Indonesia, India, and Pakistan. Correlations with epidemiological information further support the usefulness of PFGE as an epidemiological tool in laboratory investigations of suspected outbreaks. Standardization of PFGE methodology will allow international comparison of fingerprint patterns and will form the basis of a laboratory network for tracking V. cholerae.

Bacterial Typing Techniques↗
PubMed2001

Trends in multidrug-resistant Mycobacterium tuberculosis in relation to sputum smear positivity in Hong Kong, 1989-1999.

We studied retrospectively the territory-wide occurrence and trends of multidrug-resistant tuberculosis (MDR-TB) in Hong Kong over an 11-year period during which a short-course directly observed therapy ("DOTS-Plus") strategy has been in operation. The overall MDR rate was 2.1% (primary, 1.4% and acquired, 9.5%) and declined at 0.08% per year: smear-positive primary MDR cases declined at yearly rate of -0.065% (R2=.23), and smear-negative primary MDR cases increased at 0.035% yearly. With declining rates of smear positivity, sputum culture has become the mainstay of detection of MDR-TB. Although the overall notification rates showed the elderly (age >65 years) age group to be most affected, the occurrence of MDR-TB has been consistently highest in the 35-65 year age group (60.4% of MDR caseload). We demonstrated declining rates of sputum smear positivity of MDR-TB in a DOTS-plus environment and that a centralized laboratory database is essential in gathering epidemiological information for identifying particular risk groups and monitoring trends of MDR-TB in a community.

Age Distribution↗