[show abstract][hide abstract] ABSTRACT: The National University Hospital (NUH) was the first restructured public hospital in Singapore. As the most recently established hospital in Singapore, it has a unique record of alert organisms including methicillin-resistant Staphylococcus aureus (MRSA).
We performed a critical review of multiple data sources including surveillance reports, task force reports, published abstracts and manuscripts concerning MRSA in NUH.
Three themes emerged: 1) the MRSA rates have remained relatively stable through the life of the hospital despite the increased complexity of patients and intermittent intensified control efforts; 2) the major MRSA task forces were driven by surgeons and 3) a scientific approach to epidemiology has a critical role in understanding and planning interventions.
Although containment of MRSA can be accomplished to a certain degree through mobilisation of existing resources, higher goals such as eradication would require massive infusions of infrastructural, scientific and human resources to have a chance of success.
Annals of the Academy of Medicine, Singapore 11/2008; 37(10):855-60. · 1.36 Impact Factor
[show abstract][hide abstract] ABSTRACT: To assess the efficacy of screening stools sent for Clostridium difficile cytotoxin assay (CDTA) for surveillance of vancomycin-resistant enterococci (VRE).
From April to May 2005, all stools submitted for CDTA were also cultured for VRE using vancomycin containing culture media. Isolates were identified to species level and vancomycin resistance confirmed, followed by polymerase chain reaction (PCR) for detection of vancomycin resistance genes and DNA fingerprinting. Over 2 consecutive days during that period, stool specimens or rectal swabs were also obtained from all patients in high-risk units (haematology, oncology, renal and intensive care). Fifty-one patients in each group were compared in terms of VRE risk factors previously identified.
The prevalence of VRE in both groups was similar [3/204 (1.5%) in the CDTA arm and 1/97 (1.0%) in the high-risk arm; P = 1.0, Fisher's exact test]. Prevalence of risk factors for VRE colonisation, including age, duration of hospitalisation, exposure to antibiotics, exposure to surgical procedures, presence of malignancy and diabetes mellitus was similar in both groups (P > 0.05). Only renal failure (P < 0.05) was more common in the high-risk group. All 4 isolates of VRE identified were genetically distinct by variable number tandem repeat (VNTR) typing; 3 were Enterococcus faecium (2 with the vanB gene, 1 with vanA) and one E. faecalis.
Less than 2% of our high-risk patients are VRE carriers. In-hospital VRE screening using stools sent for CDTA is a simple, reasonable surrogate for screening individual high-risk patients as the patient risk profile is similar and the yield comparable in a low-prevalence setting.
Annals of the Academy of Medicine, Singapore 12/2007; 36(11):926-9. · 1.36 Impact Factor
[show abstract][hide abstract] ABSTRACT: An 18-month epidemiologic investigation of Candida bloodstream infections in a Singapore hospital identified 52 candidemic patients: 36% of whose infections were caused by C. tropicalis, 29% were due to C. albicans, 10% with C. parapsilosis and 21% involved C. glabrata. A predominant clonal C. tropicalis strain was demonstrated. No association with ICU stay, prior exposure to fluconazole/broad-spectrum antibiotics or increased mortality was found in this apparent shift towards non-C. albicans Candida species as the primary agents of candidemia.
Medical Mycology 09/2007; 45(5):435-9. · 1.98 Impact Factor
[show abstract][hide abstract] ABSTRACT: Klebsiella pneumoniae causes common and severe hospital- and community-acquired infections with a high incidence of multidrug resistance. The emergence and spread of beta-lactamase-producing K. pneumoniae strains highlight the need to develop new therapeutic strategies. In this study, we developed antisense peptide nucleic acids (PNAs) conjugated to the (KFF)(3)K peptide and investigated whether they could mediate gene-specific antisense effects in K. pneumoniae. No outer membrane permeabilization was observed with antisense PNAs when used alone. Antisense peptide-PNAs targeted at two essential genes, gyrA and ompA, were found to be growth inhibitory at concentrations of 20 muM and 40 muM, respectively. Mismatched antisense peptide-PNAs with sequence variations of the gyrA and ompA genes when used as controls were not growth inhibitory. Bactericidal effects exerted by peptide-anti-gyrA PNA and peptide-anti-ompA PNA on cells were observed within 6 h of treatment. The antisense peptide-PNAs specifically inhibited expression of DNA gyrase subunit A and OmpA from the respective targeted genes in a dose-dependent manner. Both antisense peptide-PNAs cured IMR90 cell cultures that were infected with K. pneumoniae (10(4) CFU) in a dose-dependent manner without any noticeable toxicity to the human cells.
Antimicrobial Agents and Chemotherapy 04/2007; 51(3):805-11. · 4.57 Impact Factor
[show abstract][hide abstract] ABSTRACT: Enterovirus 71 (EV71) is one of the main causative agents of hand, foot and mouth disease (HFMD) in young children. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. Thus, rapid detection of the virus is required to enable measures to be implemented in preventing widespread transmission. Based on primers and probes targeting at the VP1 region, a real-time reverse-transcriptase polymerase chain reaction (RT-PCR) hybridization probe assay was developed for specific detection of EV71 from clinical specimens. Quantitative analysis showed that the assay was able to detect as low as 5 EV71 viral copies and EV71 was detected from 46 of the 55 clinical specimens obtained from pediatric patients suffering from HFMD during the period from 2000 to 2003 in Singapore. This study showed that the single tube real-time RT-PCR assay developed in this study can be applied as a rapid and sensitive method for specific detection of EV71 directly from clinical specimens.
Molecular and Cellular Probes 05/2006; 20(2):135-40. · 1.87 Impact Factor
[show abstract][hide abstract] ABSTRACT: Klebsiella pneumoniae is an opportunistic pathogen which causes pneumoniae, urinary tract infections and septicemia in immunocompromised patients. Hospital outbreaks of multidrug-resistant K. pneumoniae, especially those in neonatal wards, are often caused by strains producing the extended-spectrum-beta-lactamases (ESBLs). An immunoproteome based approach was developed to identify candidate antigens of K. pneumoniae for vaccine development. Sera from patients with acute K. pneumoniae infections (n = 55) and a control group of sera from healthy individuals (n = 15) were analyzed for reactivity by Western blot against ESBL K. pneumoniae outer membrane proteins separated by 2-DE. Twenty highly immunogenic protein spots were identified by immunoproteomic analysis. The immunogenic proteins that are most frequently recognized by positive K. pneumoniae sera were OmpA, OmpK36, FepA, OmpK17, OmpW, Colicin I receptor protein and three novel proteins. Two of the vaccine candidate genes, OmpA (Struve et al. Microbiology 2003, 149, 167-176) and FepA (Lai, Y. C. et al.. Infect Immun 2001, 69, 7140-7145), have recently been shown to be essential in colonization and infection in an in vivo mouse model. Hence, these two immunogenic proteins could serve as potential vaccine candidates.
[show abstract][hide abstract] ABSTRACT: A rapid real-time polymerase chain reaction (PCR) assay using molecular beacons has been developed for the simultaneous detection of wild-type and mutant strains of cytomegaloviruses (CMV) with respect to codon 460 of the UL97 gene has been developed. The molecular beacons were designed to complement the wild-type codon 460 or the mutant sequence arising from a single base-pair difference (point mutation). Discrimination between wild-type and mutant templates was demonstrated as the beacons did not generate fluorescence with their respective mismatch targets but only with those that they were designed to perfectly anneal with. Samples that harbor mixed populations of CMV could also be readily recognized. Applied to a small number of clinical samples, the retrospective screening by this assay are in general concordance with that obtained by PCR-RFLP. Using molecular beacons strategy, codon 460 mutation was detected in ten out o the total number of 40 samples, whereas the latter method identified nine samples as containing the mutation. The discrepant result arose from the genotyping of one clinical sample as mixed (containing both wild-type and mutant CMV strains) by molecular beacons but as wild-type by PCR-RFLP, suggesting that this real-time strategy is possibly more sensitive for mutation analysis.
Molecular and Cellular Probes 01/2006; 19(6):389-93. · 1.87 Impact Factor
[show abstract][hide abstract] ABSTRACT: A PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed for direct identification of Pseudomonas aeruginosa from positive BACTEC blood culture bottles. PCR primers were designed to target a 249 bp sequence of the oprI gene in P. aeruginosa. Biotin-labeled probe (PA3) targeted to the species-specific motif were hybridized to the digoxigenin-labeled amplified products from P. aeruginosa and captured on streptavidin-coated microtiter plates. The specificity of the assay using the PA3 probe was investigated with a range of microorganisms, which are commonly isolated from blood culture bottles serving as negative controls. The PCR-ELISA assay was shown to be highly specific for the identification of P. aeruginosa and was 10-fold more sensitive than an agarose gel-based detection method using the same pair of primers, with a detection limit at 10 fg of template. The PCR-ELISA assay developed in this study is 100% sensitive and 100% specific as it correctly identified all 73 positive and 42 negative controls as well as 25 double blind clinical samples. It significantly reduces the time needed for the identification of P. aeruginosa from positive BACTEC blood cultures bottles from 2-3 days to 6-8h.
Molecular and Cellular Probes 01/2006; 19(6):417-21. · 1.87 Impact Factor
[show abstract][hide abstract] ABSTRACT: A LightCycler real-time PCR hybridization probe-based assay which detects a partial Klebsiella pneumoniae 16S rRNA gene was developed for the rapid identification of K. pneumoniae directly from growth-positive blood culture bottles (BACTEC 9240 system) within 2 h. No cross-reactivity was observed with 65 negative-control blood cultures that grew bacteria other than K. pneumoniae and 48 negative blood cultures from double-blind experiments, thus demonstrating 100% specificity when compared to results of conventional biochemical characterization. The assay also showed 100% sensitivity, as it correctly identified all 142 positive-control blood cultures and 4 from double-blind trials.
Journal of Clinical Microbiology 04/2004; 42(3):1337-40. · 4.07 Impact Factor
[show abstract][hide abstract] ABSTRACT: Several nucleic acid amplification (NAA) tests for Mycobacterium tuberculosis (MTB) have been licensed for the rapid diagnosis of active pulmonary tuberculosis (PTB) in respiratory secretions. There is uncertainty however regarding the practical application of these tests in clinical decision making.
To evaluate the utility of the COBAS AMPLICOR assay (Roche Diagnostics; Singapore) for MTB as applied by specialists for the rapid diagnosis of PTB in the routine clinical setting.
A prospective study of consecutive patients suspected of PTB and tested with the AMPLICOR assay under the care of respiratory physicians. The final diagnosis was based on all relevant clinical information after at least 3 months of follow-up. Accuracy of the NAA test was compared with that of the initial expectant treatment. Expectant treatment was based on an integrated approach that incorporated clinical evaluation with results of direct smear and NAA tests.
The incidence of PTB in 168 patients was 32%. The basis for expectant treatment of PTB was positive smear result in 47%, clinical suspicion in 26%, and positive AMPLICOR result in 23%. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the AMPLICOR test were 77%, 100%, 99%, 90%, and 93%, respectively. In comparison, they were 96%, 97%, 94%, 98%, and 97%, respectively, for the integrated clinical approach.
In the rapid diagnosis of PTB, the clinical judgment of specialists augmented the utility of the NAA test: (1) specialists selected patients with high-to-moderate pretest probabilities, (2) they commenced treatment promptly on a positive NAA test result, and (3) they were willing to start treatment in some patients on the basis of high clinical suspicion despite negative smear and negative NAA test results.
[show abstract][hide abstract] ABSTRACT: To assess the frequency of community-acquired methicillin-resistant Staphylococcus aureus (MRSA) infections.
A teaching hospital in Singapore.
Prospectively collected surveillance data were reviewed during a 1-year period to determine the extent and origin of community-acquired MRSA infections.
Whereas 32% of 383 MRSA infections were detected less than 48 hours after hospital admission and would, by convention, be classified as "community acquired," all but one of these were among patients who had been exposed to outpatient centers including dialysis or chemotherapy clinics, visiting nurses, community hospitals, or all three.
With health care increasingly being delivered in an outpatient setting, community-acquired MRSA infections are often acquired in hospital-related sites and most may be more accurately described as "healthcare acquired." Infection control measures need to move beyond the traditional paradigm of acute care hospitals to effectively control the spread of resistant pathogens.
Infection Control and Hospital Epidemiology 07/2003; 24(6):436-8. · 4.02 Impact Factor
[show abstract][hide abstract] ABSTRACT: The aim of this study was to determine the most appropriate strategy for the rapid diagnosis of pulmonary tuberculosis (PTB) using a nucleic acid amplification (NAA) test.
This was a prospective study of 128 adult patients in whom respiratory secretions were tested for Mycobacterium tuberculosis by the AMPLICOR assay. The basis for starting PTB treatment was noted for each patient. The optimal approach was determined by using Bayes' theorem to compare different combinations of pretest probability, smear results with the AMPLICOR test.
The incidence of PTB was 15.6%. In only one patient was treatment for PTB commenced because of a positive AMPLICOR result. The rest were managed according to the conventional approach which relied upon clinical judgment and direct smear. The optimal approach was to treat patients with high or intermediate pretest risk for PTB who returned positive AMPLICOR tests. The overall accuracies of the conventional approach, AMPLICOR test and optimal approach were 89.8, 95.3 and 96.1%, respectively.
This small study suggests that NAA testing be limited to patients with high or intermediate pretest risk of PTB. In this group, positive results demand treatment while the management of those with negative results still relies on clinical judgment.
[show abstract][hide abstract] ABSTRACT: Several nucleic acid amplification (NAA) tests are available for the rapid detection of Mycobacterium tuberculosis (MTB) in clinical specimens.
To identify the pattern of utilisation and accuracy of the AMPLICOR test in routine clinical practice in an acute care setting.
A retrospective descriptive study.
We studied 159 consecutive specimens in which the AMPLICOR (Roche; Branchburg, NJ) test was requested by attending doctors. The sensitivities and specificities of the AMPLICOR for detection of active tuberculosis (TB) were calculated in relation to types of specimens, smear and culture results.
The number of requests more than doubled from 1999 to 2000. Thirty-eight percent of the specimens were not from the respiratory tract. The majority of the specimens had requests for one or more additional test (mean 1.8). The rate of active TB was 18%. The sensitivities of the AMPLICOR on per specimen, per patient, per smear negative specimen and per smear negative patient basis were found to be 81%, 80%, 66.7% and 71.4% respectively. The specificities for these groups accordingly were 99%, 98.6%, 99% and 98.6% respectively. The sensitivity and specificity for respiratory specimens were 97.5% and 98.5%, while for non-respiratory specimens, they were 60% and 100%. In smear negative specimens, the sensitivity and specificity for respiratory specimens were 60% and 98.5%, while for non-respiratory specimens, they were 75% and 100%. The AMPLICOR assay was negative in all 21 specimens of pleural or spinal fluid.
There is a growing demand for NAA in the rapid diagnosis of TB with a high proportion of non-respiratory specimens. The number of additional diagnostic tests performed on each specimen should be limited. In routine clinical practice, the AMPLICOR assay is a useful confirmatory test for active pulmonary TB. The utility of the AMPLICOR assay for MTB detection in exudative fluid specimens needs further evaluation.
Singapore medical journal 09/2002; 43(8):415-20. · 0.63 Impact Factor
[show abstract][hide abstract] ABSTRACT: Singapore is a modern urban city and endemic typhus is thought to be a disease of the past. This may be due to lack of specific serological testing as indirect immunoperoxidase testing using specific rickettsial antigens (U.S. Army Medical Research Unit, Institute of Medical Research, Kuala Lumpur, Malaysia) has only recently become available. In the last fourteen months, twenty-one cases of endemic typhus were diagnosed in patients hospitalised for acute febrile illnesses at the National University Hospital. We conducted a case control study to define the clinical and laboratory features of endemic typhus in Singapore.
Demographic, clinical and laboratory data were reviewed for cases and twenty-one age and sex matched controls who had negative serologic tests as part of a work-up for fever of unknown origin.
Apart from a higher initial temperature (39 degrees C vs 37.9 degrees C (p < 0.001)) and ALT(p = 0.002), cases and controls had similar presentations of fever, myalgia, headache, cough, normal WBC and platelet counts. Singapore residents and migrant workers were represented in both groups (p = ns).
Endemic typhus remains an important cause of acute febrile illness in Singaporein both the local and migrant worker populations. The presentation is similar to other causes of acute febrile illnesses and the diagnosis will be missed unless it is specifically sought.
Singapore medical journal 12/2001; 42(12):549-52. · 0.63 Impact Factor
[show abstract][hide abstract] ABSTRACT: The activity of cefpirome, cefepime and piperacillin/tazobactam previously unused in the hospital was evaluated in parallel with five broad-spectrum antibiotics (ceftazidime, ceftriaxone, imipenem, ciprofloxacin and amikacin) currently being used to treat serious infections in the National University Hospital, Singapore. Two hundred and two clinically significant, organisms consecutively isolated during 1998 were included in the study. In vitro efficacy of cefepime, cefpirome and piperacillin/tazobactam was not superior to imipenem, ciprofloxacin and amikacin which are currently used. More than 40% of Enterbacteriaceae were found to be ESBL producers. The incidence of nosocomial organisms resistant to drugs used in serious infections had increased since 1995. Imipenem-resistance occurred in 34.4% of Acinetobacter spp. and 19.2% Pseudomonas aeruginosa. No single agent appeared to be suitable for empirical monotherapy of serious sepsis.
International Journal of Antimicrobial Agents 11/2001; 18(4):391-3. · 4.42 Impact Factor
[show abstract][hide abstract] ABSTRACT: The AMPLICOR assay (Roche; Branchburg, NJ), a rapid direct amplification test for Mycobacterium tuberculosis, has only been licensed for use in smear-positive respiratory specimens. However, many patients with pulmonary tuberculosis (PTB) have smear-negative disease. The clinical utility of this test in patients with smear-negative PTB is unknown.
To evaluate the effect of pretest probability of PTB estimated by chest physicians on the accuracy of the AMPLICOR assay in patients with smear-negative PTB. Design and methods: A prospective study of consecutive patients suspected of having smear-negative PTB. Two chest physicians estimated the pretest probability of active disease (high, intermediate, and low categories). Respiratory specimens were examined with radiometric broth medium cultures and with the AMPLICOR assay for M tuberculosis. The decision on a final diagnosis of PTB was blinded to the AMPLICOR results.
Active PTB was diagnosed in 25 of 441 patients (5.7%). The AMPLICOR assay had an overall sensitivity of 44% and a specificity of 99%. Results of the assay were negative in seven patients with culture-negative PTB. The proportions of patients in the high, intermediate, and low pretest groups were 4.5%, 19.7%, and 75.7%, respectively. The incidence of PTB for each group was 95%, 3.4%, and 0.9%, respectively. The sensitivities of the AMPLICOR assay in the three groups of patients were 47%, 33%, and 33%, respectively, while the specificities were 100%, 98%, and 99%, respectively.
In patients suspected of having smear-negative PTB, the following conclusions were drawn: (1) the incidence of active PTB was low; (2) pretest estimates accurately discriminated between patients with high and low risk of PTB; (3) the risk of PTB was overestimated in the intermediate group; and (4) the utility of the AMPLICOR assay in the intermediate-risk group may be limited by the overestimation of disease prevalence and low test sensitivity. Further studies are needed on the role of the AMPLICOR assay in better selected patients with an intermediate risk of having smear-negative PTB.