Comparison of PneumID real-time PCR assay with Amplex eazyplex LAMP assay for laboratory diagnosis of Pneumocystis jirovecii Pneumonia.

We compared PneumID PCR with Amplex eazyplex LAMP assay for the diagnosis of Pneumocystis jirovecii pneumonia (PJP). Both assays enable accurate diagnosis of definite PJP. Cut-off cycle threshold of the PneumID assay was < 26.68 while the cut-off time-to-positivity of the eazyplex assay was 16:02 (minutes:seconds). The positive and negative percentage agreement of eazyplex assay with PneumID assay was 75.0% and 100.0% respectively, while the overall agreement was substantial with kappa = 0.80. For both assays, establishment of cut-off values to differentiate probable PJP from colonization was not feasible as results overlapped.

Both PneumID PCR and Amplex eazyplex LAMP assay enable accurate diagnosis of definite Pneumocystis jirovecii pneumonia (PJP). PneumID assay was more sensitive than eazyplex assay for detection of P. jirovecii. However, differentiation between probable PJP from colonization was not feasible.

Surveillance of antibiotic resistance among common Clostridium difficile ribotypes in Hong Kong.

Incidence of Clostridium difficile infection (CDI) is rapidly increasing and it poses a major health burden globally. However, data regarding the epidemiology of CDI in Asia are limited. We aimed to characterize the antimicrobial susceptibility patterns of common ribotypes of toxigenic C. difficile in Hong Kong. Fifty-three PCR ribotypes were identified among 284 toxigenic C. difficile clinical isolates. The five most prevalent ribotypes were 002 (13%), 017 (12%), 014 (10%), 012 (9.2%), and 020 (9.5%). All tested C. difficile strains remained susceptible to metronidazole, vancomycin, meropenem and piperacillin/tazobactam, but highly resistant to cephalosporins. Of the fluoroquinolones, highest resistance to ciprofloxacin was observed (99%), followed by levofloxacin (43%) and moxifloxacin (23%). The two newly emerged PCR ribotypes, 017 and 002, demonstrated high levels of co-resistance towards clindamycin, tetracycline, erythromycin and moxifloxacin. PCR ribotypes 017 and 002 with multi-drug resistance are rapidly emerging and continuous surveillance is important to monitor the epidemiology of C. difficile to prevent outbreaks of CDI.

Hong Kong Chinese school children with elevated urine melamine levels: a prospective follow up study.

BACKGROUND: In 2008, the outbreak of kidney stones in children fed by melamine-tainted milk products in Mainland China has caused major public concern of food safety. We identified Hong Kong school children with elevated urine melamine level from a community-based school survey in 2007-08 and reviewed their clinical status in 2009. METHODS: In 2007-08, 2119 school children participated in a primary and secondary school survey in Hong Kong using a cluster sampling method. Urine aliquots from 502 subjects were assayed for melamine level. High urine melamine level was defined as urine melamine/creatinine ratio >7.1 µg/mmol. Subjects with high urine melamine level were invited for clinical evaluation in 2009 including urinalysis and ultrasound imaging of the urinary system. RESULTS: The age range of this subcohort was 6 - 20 years with 67% girls (335 female and 167 male subjects). The spot urine melamine/creatinine ratio of the 502 urine aliquots ranged from undetectable to 1467 µg/mmol (median 0.8 µg/mmol). Of these, 213 subjects had undetectable level (42%). We invited 47 (9%) subjects with high urine melamine level for re-evaluation and one subject declined. The median duration of follow-up was 23.5 months (interquartile range: 19.8 - 30.6 months). None of the 46 subjects (28% boys, mean age 13.9 ± 2.9 years) had any abnormality detected on ultrasound study of the urinary system. All subjects had stable renal function with a median urine albumin-creatinine ratio of 0.70 mg/mmol (interquartile range: 0.00 - 2.55 mg/mmol). CONCLUSIONS: Hong Kong Chinese school children with high urine melamine levels appeared to have benign clinical course in the short term although a long term follow-up study is advisable in those with persistently high urine melamine level.

High-level gentamicin resistance mediated by a Tn4001-like transposon in seven nonclonal hospital isolates of Streptococcus pasteurianus.

We report on the first occurrence of high-level gentamicin resistance (MICs > or = 512 microg/ml) in seven clinical isolates of Streptococcus pasteurianus from Hong Kong. These seven isolates were confirmed to be the species S. pasteurianus on the basis of nucleotide sequencing of the superoxide dismutase (sodA) gene. Epidemiological data as well as the results of pulse-field gel electrophoresis analysis suggested that the seven S. pasteurianus isolates did not belong to the same clone. Molecular characterization showed that they carried a chromosomal, transposon-borne resistance gene [aac(6')Ie-aph(2'')Ia] which was known to encode a bifunctional aminoglycoside-modifying enzyme. The genetic arrangement of this transposon was similar to that of Tn4001, a transposon previously recovered from Staphylococcus aureus and other gram-positive isolates. Genetic linkage with other resistance elements, such as the ermB gene for erythromycin resistance, was not evident. On the basis of these findings, we suggest that routine screening for high-level gentamicin resistance should be recommended for all clinically significant blood culture isolates. This is to avoid the inadvertent use of short-course combination therapy with penicillin and gentamicin, which may lead to the failure of treatment for endocarditis, the selection of drug-resistant Streptococcus pasteurianus and other gram-positive organisms, as well as the unnecessary usage of gentamicin, a drug with potential toxicity.

Persistent Staphylococcus capitis septicemia in a preterm infant.

A preterm infant had persistent Staphylococcus capitis septicemia with 11 consecutive positive blood cultures over a period of 33 days. The clinical evidence suggested that the source of infection probably originated from the gastrointestinal tract. The combination of rifampin and linezolid treatment, together with prolonged stoppage of enteral feeding, successfully terminated the infection. Rifampin and linezolid should be considered as alternative antimicrobial agents when glycopeptides fail to eradicate Gram-positive pathogens from the host.

Psychrobacter bacteraemia in a cirrhotic patient after the consumption of raw geoduck clam.

Psychrobacter species rarely cause infections in human. We reported herein a 62-year-old cirrhotic patient who presented with fever and diarrhoea after the consumption of raw geoduck clam. Blood culture grew Psychrobacter phenylpyruvicus which was sensitive to most anti-microbial agents. The patient responded promptly to intravenous antibiotics. This is the first report of human infection with this unusual organism which illustrates the potential health hazards of eating raw geoduck clam in patients with advanced liver disease.

Extent and predictors of microbial hand contamination in a tertiary care ophthalmic outpatient practice.

PURPOSE: To measure the extent of microbial hand contamination among ophthalmologists during routine clinic practice and examine its association with hand cleansing practices and beliefs, glove use, and patient load. METHODS: This was a single-masked analysis of resident and transient flora of ophthalmologists before and after patient examination and after handwashing by agar imprints of the dominant hand. Standardized questionnaires were used to collect information concerning subjects' hand cleansing practices and patient load. RESULTS: Of the 108 cultures, 107 (99.1%) were culture positive, yielding 15 separate organisms. Gram-negative bacilli were the most common transient flora, followed by Gram-positive cocci and fungi. Thirty-five (97.2%) ophthalmologists were culture positive for at least one resident and 8 (22.2%) ophthalmologists were culture positive for at least one transient organism, before patient contact. Regression models showed alcohol-based hand rub use, transient and resident floral load before patient contact, and patient load collectively accounted for 58.7% of the variance in resident floral load after patient contact. Use of alcohol-based hand rubs was associated with a mean resident floral reduction of 324.4 CFUs (95% confidence interval [CI] = 185.4 to 463.5; P < 0.01) and 31.6 CFUs (95% CI = 1.2 to 62.0; P < 0.05) after patient contact and handwashing, respectively. Handwashing with chlorhexidine was a significant predictor for transient floral load after handwashing (unstandardized beta = -17.2; 95% CI = -10.2 to -24.2; P < 0.01). CONCLUSIONS: The extent of contamination with pathogenic organisms after contact with eye outpatients, who have traditionally been perceived as relatively "clean," was of concern. Previously identified risk factors for hand contamination in inpatient settings, such as patient load, only explained a small proportion of variance in microbial load in the ophthalmic outpatient setting.

Immunofluorescence assay for serologic diagnosis of SARS.

We evaluated an indirect immunofluorescence assay based on virus-infected cells for detecting anti-severe acute respiratory syndrome-associated coronavirus (SARS-CoV) immunoglobulin (Ig) G antibody. All confirmed SARS cases demonstrated seroconversion or fourfold rise in IgG antibody titer; no control was positive. Sensitivity and specificity of this assay were both 100%. Immunofluorescence assay can ascertain the status of SARS-CoV infection.