A multicenter study was conducted to evaluate the diagnostic performance and the time to identifcation of the Verigene Blood Culture Test, the BC-GP and BC-GN assays, to identify both Gram-positive ...and Gram-negative bacteria and their drug resistance determinants directly from positive blood cultures collected in Hong Kong.
A total of 364 blood cultures were prospectively collected from four public hospitals, in which 114 and 250 cultures yielded Gram-positive and Gram-negative bacteria, and were tested with the BC-GP and BC-GN assay respectively. The overall identification agreement for Gram-positive and Gram-negative bacteria were 89.6% and 90.5% in monomicrobial cultures and 62.5% and 53.6% in polymicrobial cultures, respectively. The sensitivities for most genus/species achieved at least 80% except Enterococcus spp. (60%), K.oxytoca (0%), K.pneumoniae (69.2%), whereas the specificities for all targets ranged from 98.9% to 100%. Of note, 50% (7/14) cultures containing K.pneumoniae that were missed by the BC-GN assay were subsequently identified as K.variicola. Approximately 5.5% (20/364) cultures contained non-target organisms, of which Aeromonas spp. accounted for 25% and are of particular concern. For drug resistance determination, the Verigene test showed 100% sensitivity for identification of MRSA, VRE and carbapenem resistant Acinetobacter, and 84.4% for ESBL-producing Enterobacteriaceae based on the positive detection of mecA, vanA, blaOXA and blaCTXM respectively.
Overall, the Verigene test provided acceptable accuracy for identification of bacteria and resistance markers with a range of turnaround time 40.5 to 99.2 h faster than conventional methods in our region.
Hong Kong experienced a surge of Omicron BA.2 infections in early 2022, resulting in one of the highest per-capita death rates of COVID-19. The outbreak occurred in a dense population with low ...immunity towards natural SARS-CoV-2 infection, high vaccine hesitancy in vulnerable populations, comprehensive disease surveillance and the capacity for stringent public health and social measures (PHSMs). By analyzing genome sequences and epidemiological data, we reconstructed the epidemic trajectory of BA.2 wave and found that the initial BA.2 community transmission emerged from cross-infection within hotel quarantine. The rapid implementation of PHSMs suppressed early epidemic growth but the effective reproduction number (R
) increased again during the Spring festival in early February and remained around 1 until early April. Independent estimates of point prevalence and incidence using phylodynamics also showed extensive superspreading at this time, which likely contributed to the rapid expansion of the epidemic. Discordant inferences based on genomic and epidemiological data underscore the need for research to improve near real-time epidemic growth estimates by combining multiple disparate data sources to better inform outbreak response policy.
Molecular methods for the detection of drug-resistant tuberculosis are potentially more rapid than conventional culture-based drug susceptibility testing, facilitating the commencement of appropriate ...treatment for patients with drug resistant tuberculosis. We aimed to develop and evaluate high-resolution melting (HRM) assays for the detection of mutations within gyrA, rpsL, and rrs, for the determination of fluoroquinolone and streptomycin resistance in Mycobacterium tuberculosis (MTB).
A blinded series of DNA samples extracted from a total of 92 clinical isolates of MTB were analyzed by HRM analysis, and the results were verified using DNA sequencing. The sensitivity and specificity of the HRM assays in comparison with drug susceptibility testing were 74.1% and 100.0% for the detection of fluoroquinolone resistance, and 87.5% and 100.0% for streptomycin resistance. Five isolates with low level resistance to ofloxacin had no mutations detected in gyrA, possibly due to the action of efflux pumps, or false negativity due to mixed infections. One fluoroquinolone-resistant isolate had a mutation in a region of gyrA not encompassed by our assay. Six streptomycin-resistant strains had undetectable mutations by HRM and DNA sequencing, which may be explained by the fact that not all streptomycin-resistant isolates have mutations within rpsL and rrs, and suggesting that other targets may be involved.
The HRM assays described here are potentially useful adjunct tests for the efficient determination of fluoroquinolone and streptomycin resistance in MTB, and could facilitate the timely administration of appropriate treatment for patients infected with drug-resistant TB.
Loop-mediated isothermal amplification (LAMP) has received considerable attention for decentralized (point-of-care and on-site) nucleic acid testing in view of its simple temperature control (60–65 ...°C) and short assay time (15–60 min). There remains a challenge in its wide adoption and acceptance due to the limitations of the existing amplification result reporter probes, e.g., photobleaching of organic fluorophore and reduced sensitivity of the pH-sensitive colorimetric dye. Herein, we demonstrate CdSeS/ZnS quantum dots (semiconductor fluorescent nanocrystals with superior photostability than organic fluorophore) with surface modification of cysteamine (amine-QDs) as a new reporter probe for LAMP that enabled single-copy sensitivity (limit of detection of 83 zM; 20 μL reaction volume). For a negative LAMP sample (absence of target sequence), positively charged amine-QDs remained dispersed due to interparticle electrostatic repulsion. While for a positive LAMP sample (presence of target sequence), amine-QDs became precipitated. The characterization data showed that amine-QDs were embedded in magnesium pyrophosphate crystals (generated during positive LAMP), thus leading to their coprecipitation. This amine-QD-based one-step LAMP assay advances the field of QD-based nucleic acid amplification assays in two aspects: (1) compatibilityone-step amplification and detection (versus separation of amplification and detection steps); and (2) universalitythe same amine-QDs for different target sequences (versus different oligonucleotide-modified QDs for different target sequences).
Proper drying of hands after washing is an integral part of hand hygiene. An experimental study on 30 subjects using multiple comparisons of six hand drying methods including 1) drying on own ...clothes, 2) drying with one paper towel, 3) drying with two paper towels, 4) drying with a warm air dryer while holding hands stationary for 20 s, 5) drying with a warm air dryer while hand rubbing for 20 s, and 6) drying with a jet air dryer until complete dryness was achieved. It aimed to determine the effectiveness of different hand drying methods for removing bacteria from washed hands, so as to identify the optimum method using minimum resources. Our study demonstrated that the use of jet air dryers is the best method to eliminate bacteria on hands, whereas drying hands on one's own clothes is the least effective. Drying hands in a stationary position could remove more bacteria than rubbing hands when using a warm air dryer for 20 s, which mimics people's usual hand-drying practice. No significant difference in bacteria reduction was detected between the use of one or two paper towels for hand drying; therefore, using fewer resources is recommended to maintain environmental sustainability.
We have developed a high-resolution melting (HRM) assay to scan for mutations in the rpoB, inhA, ahpC, and katG genes and/or promoter regions for the detection of rifampin and isoniazid resistance in ...Mycobacterium tuberculosis. For assay development, 23 drug-resistant isolates of M. tuberculosis having 29 different mutations, together with 40 drug-susceptible isolates, were utilized. All 29 mutations were accurately detected by our assay. We further validated the assay with a series of 59 samples tested in a blind manner. All sequence alterations that were within the regions targeted by the HRM assay were correctly identified. Compared against results of DNA sequencing, the sensitivity and specificity of our HRM assay were 100%. For the blinded samples, the specificities and sensitivities were 89.3% and 100%, respectively, for detecting rifampin resistance and 98.1% and 83.3%, respectively, for detecting isoniazid resistance, as isolates with mutations in regions not encompassed by our assay were not detected. A C-to-T sequence alteration at position -15 of the ahpC regulatory region, which was previously reported to be associated with isoniazid resistance, may possibly be a polymorphism, as it was detected in an isoniazid-susceptible M. tuberculosis isolate. HRM is a rapid, accurate, simple, closed-tube, and low-cost method. It is thus an ideal assay to be used in countries with a high prevalence of drug-resistant M. tuberculosis and where cost-effectiveness is essential. As a mutation-scanning assay for detecting drug-resistant M. tuberculosis, it can potentially lead to better treatment outcomes resulting from earlier treatment with the appropriate antibiotics.
Many people use handwashing and hand-drying facilities in public washrooms under the impression that these amenities are hygienic. However, such facilities may be potential sites for the transmission ...of pathogenic bacteria. This study aimed to examine the hygiene facilities provided including handwashing and hand-drying facilities in public washrooms. Total bacterial counts and species identification were determined for hand-drying facilities. Antimicrobial susceptibilities were performed.
The bacterial contamination levels of 55 public washrooms ranging in category from low class communities to high end establishments, were examined. The hygienic environment and facilities of the washrooms were analysed using an electronic checklist to facilitate immediate data entry. Pre-moistened sterile swabs were used to collect samples from areas around the outlet of paper towel dispensers, air outlet of air dryers, exit door handles and paper towels in the washrooms. Total bacterial counts were performed and isolates identified using matrix-assisted laser desorption ionisation time-of-flight mass spectrometry. Antimicrobial susceptibility was determined by disk diffusion.
The high and middle-income categories washrooms generally had cleaner facilities and environment followed by those in low categories. Fifty-two bacterial species were identified from the 55 investigated washrooms. Over 97% of the pathogenic
tested were resistant to at least one first-line antimicrobial therapeutic agent, including penicillin, cefoxitin, erythromycin, co-trimoxazole, clindamycin and gentamicin, and 22.6% demonstrated co-resistance to at least three antimicrobial agents, with co-resistance to penicillin, erythromycin and clindamycin being the most common.
Our findings suggest that hand-drying facilities in public washrooms can act as reservoirs of drug-resistant bacteria. The importance of frequent cleaning and maintenance of public washrooms to promote safe hand hygiene practices for the public are emphasised.
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