Transmission Patterns of Co-Circulation of Omicron Sub-Lineages in Hong Kong SAR, China, a City with Rigorous Social Distancing Measures, in 2022.

OBJECTIVE: This study aimed to characterize the changing landscape of circulating SARS-CoV-2 lineages in the local community of Hong Kong throughout 2022. We examined how adjustments to quarantine arrangements influenced the transmission pattern of Omicron variants in a city with relatively rigorous social distancing measures at that time. METHODS: In 2022, a total of 4684 local SARS-CoV-2 genomes were sequenced using the Oxford Nanopore GridION sequencer. SARS-CoV-2 consensus genomes were generated by MAFFT, and the maximum likelihood phylogeny of these genomes was determined using IQ-TREE. The dynamic changes in lineages were depicted in a time tree created by Nextstrain. Statistical analysis was conducted to assess the correlation between changes in the number of lineages and adjustments to quarantine arrangements. RESULTS: By the end of 2022, a total of 83 SARS-CoV-2 lineages were identified in the community. The increase in the number of new lineages was significantly associated with the relaxation of quarantine arrangements (One-way ANOVA, F(5, 47) = 18.233, p < 0.001)). Over time, Omicron BA.5 sub-lineages replaced BA.2.2 and became the predominant Omicron variants in Hong Kong. The influx of new lineages reshaped the dynamics of Omicron variants in the community without fluctuating the death rate and hospitalization rate (One-way ANOVA, F(5, 47) = 2.037, p = 0.091). CONCLUSION: This study revealed that even with an extended mandatory quarantine period for incoming travelers, it may not be feasible to completely prevent the introduction and subsequent community spread of highly contagious Omicron variants. Ongoing molecular surveillance of COVID-19 remains essential to monitor the emergence of new recombinant variants.

Probable airborne transmission of Burkholderia pseudomallei causing an urban outbreak of melioidosis during typhoon season in Hong Kong, China.

Between January 2015 and October 2022, 38 patients with culture-confirmed melioidosis were identified in the Kowloon West (KW) Region, Hong Kong. Notably, 30 of them were clustered in the Sham Shui Po (SSP) district, which covers an estimated area of 2.5 km2. Between August and October 2022, 18 patients were identified in this district after heavy rainfall and typhoons. The sudden upsurge in cases prompted an environmental investigation, which involved collecting 20 air samples and 72 soil samples from residential areas near the patients. A viable isolate of Burkholderia pseudomallei was obtained from an air sample collected at a building site five days after a typhoon. B. pseudomallei DNA was also detected in 21 soil samples collected from the building site and adjacent gardening areas using full-length 16S rRNA gene sequencing, suggesting that B. psuedomallei is widely distributed in the soil environment surrounding the district. Core genome-multilocus sequence typing showed that the air sample isolate was phylogenetically clustered with the outbreak isolates in KW Region. Multispectral satellite imagery revealed a continuous reduction in vegetation region in SSP district by 162,255 m2 from 2016 to 2022, supporting the hypothesis of inhalation of aerosols from the contaminated soil as the transmission route of melioidosis during extreme weather events. This is because the bacteria in unvegetated soil are more easily spread by winds. In consistent with inhalational melioidosis, 24 (63.2%) patients had pneumonia. Clinicians should be aware of melioidosis during typhoon season and initiate appropriate investigation and treatment for patients with compatible symptoms.

The clinical utility of Nanopore 16S rRNA gene sequencing for direct bacterial identification in normally sterile body fluids.

The prolonged incubation period of traditional culture methods leads to a delay in diagnosing invasive infections. Nanopore 16S rRNA gene sequencing (Nanopore 16S) offers a potential rapid diagnostic approach for directly identifying bacteria in infected body fluids. To evaluate the clinical utility of Nanopore 16S, we conducted a study involving the collection and sequencing of 128 monomicrobial samples, 65 polymicrobial samples, and 20 culture-negative body fluids. To minimize classification bias, taxonomic classification was performed using 3 analysis pipelines: Epi2me, Emu, and NanoCLUST. The result was compared to the culture references. The limit of detection of Nanopore 16S was also determined using simulated bacteremic blood samples. Among the three classifiers, Emu demonstrated the highest concordance with the culture results. It correctly identified the taxon of 125 (97.7%) of the 128 monomicrobial samples, compared to 109 (85.2%) for Epi2me and 102 (79.7%) for NanoCLUST. For the 230 cultured species in the 65 polymicrobial samples, Emu correctly identified 188 (81.7%) cultured species, compared to 174 (75.7%) for Epi2me and 125 (54.3%) for NanoCLUST. Through ROC analysis on the monomicrobial samples, we determined a threshold of relative abundance at 0.058 for distinguishing potential pathogens from background in Nanopore 16S. Applying this threshold resulted in the identification of 107 (83.6%), 117 (91.4%), and 114 (91.2%) correctly detected samples for Epi2me, Emu, and NanoCLUST, respectively, in the monomicrobial samples. Nanopore 16S coupled with Epi2me could provide preliminary results within 6 h. However, the ROC analysis of polymicrobial samples exhibited a random-like performance, making it difficult to establish a threshold. The overall limit of detection for Nanopore 16S was found to be about 90 CFU/ml.

Induction of amphotericin B resistance in susceptible Candida auris by extracellular vesicles.

Drug resistance derived from extracellular vesicles (EVs) is an increasingly important research area but has seldom been described regarding fungal pathogens. Here, we characterized EVs derived from a triazole-resistant but amphotericin B-susceptible strain of Candida auris. Nano- to microgram concentrations of C. auris EVs prepared from both broth and solid agar cultures could robustly increase the yeast's survival against both pure and clinical amphotericin B formulations in a dose-dependent manner, resulting in up to 16-fold changes of minimum inhibitory concentration. Meanwhile, this effect was not observed upon addition of these EVs to C. albicans, nor upon addition of C. albicans EVs to C. auris. No change in susceptibilities was observed upon EV treatment for fluconazole, voriconazole, micafungin, and flucytosine. Mass spectrometry indicated the presence of immunogenic-/drug resistance-implicated proteins in C. auris EVs, including alcohol dehydrogenase 1 as well as C. albicans Mp65-like and Xog1-like proteins in high quantities. Based on these observations, we propose a potential species-specific role for EVs in amphotericin B resistance in C. auris. These observations may provide critical insights into treatment of multidrug-resistant C. auris.

Co-circulation of two SARS-CoV-2 variant strains within imported pet hamsters in Hong Kong.

During the investigation of a pet shop outbreak of severe acute respiratory coronavirus 2 (SARS-CoV-2) with probable hamster-to-human transmission, the environmental and hamster samples in epidemiologically linked pet shops were found positive for SARS-CoV-2 Delta variant AY.127 strains which are phylogenetically closely related to patients and reported European strains. This interspecies' spill-over has triggered transmission in 58 patients epidemiologically linked to three pet shops. Incidentally, three dwarf hamsters imported from the Netherlands and centralized in a warehouse distributing animals to pet shops were positive for SARS-CoV-2 spike variant phylogenetically related to European B.1.258 strains from March 2020. This B.1.258 strain almost disappeared in July 2021. While no hamster-to-human transmission of B.1.258-like strain was found in this outbreak, molecular docking showed that its spike receptor-binding domain (RBD) has a similar binding energy to human ACE2 compared to that of Delta variant AY.127. Therefore, the potential of this B.1.258-related spike variant for interspecies jumping cannot be ignored. The co-circulation of B.1.258-related spike variants with Delta AY.127, which originated in Europe and was not previously found in Hong Kong, suggested that hamsters in our wholesale warehouse and retail pet shops more likely have acquired these viruses in the Netherlands or stopovers during delivery by aviation than locally. The risk of human-to-hamster reverse zoonosis by multiple SARS-CoV-2 variants leading to further adaptive spike mutations with subsequent transmission back to humans cannot be underestimated as an outbreak source of COVID-19. Testing imported pet animals susceptible to SARS-CoV-2 is warranted to prevent future outbreaks.

A low-cost TaqMan minor groove binder probe-based one-step RT-qPCR assay for rapid identification of N501Y variants of SARS-CoV-2.

The increasing prevalence of N501Y variants of SARS-CoV-2 has kindled global concern due to their enhanced transmissibility. Genome sequencing is the gold standard method to identify the emerging variants of concern. But it is time-consuming and expensive, limiting the widespread deployment of genome surveillance in some countries. Health authorities surge the development of alternative assay to expand screening capacity with reduced time and cost. In this study, we developed an in-house TaqMan minor groove binder (MGB) probe-based one-step RT-qPCR assay to detect the presence of N501Y mutation in SARS-CoV-2. A total of 168 SARS-CoV-2 positive respiratory specimens were collected to determine diagnostic accuracy of the RT-qPCR assay. As a reference standard, PANGO lineages and the mutation patterns of all samples were characterised by whole-genome sequencing. The analytical sensitivity and the ability of the assay to detect low frequency of N501Y variants were also evaluated. A total of 31 PANGO lineages were identified from 168 SARS-CoV-2 positive cases, in which 34 samples belonged to N501Y variants, including B.1.1.7 (n = 20), B.1.351 (n = 12) and P.3 (n = 2). The N501Y RT-qPCR correctly identified all 34 samples as N501Y-positive and the other 134 samples as wildtype. The limit-of-detection of the assay consistently achieved 1.5 copies/µL on four different qPCR platforms. N501Y mutation was successfully detected at an allele frequency as low as 10 % in a sample with mixed SARS-CoV-2 lineage. The N501Y RT-qPCR is simple and inexpensive (US$1.6 per sample). It enables robust high-throughput screening for surveillance of SARS-CoV-2 variants of concern harbouring N501Y mutation.

Invasive Burkholderia cepacia Complex Infections among Persons Who Inject Drugs, Hong Kong, China, 2016-2019.

During March 2016-January 2019, Burkholderia cepacia complex (BCC) infection developed in 13 persons who inject drugs (PWID) in Kowloon West Region, Hong Kong, China. Seven cases were infective spondylitis, 2 endocarditis, 2 septic arthritis, 1 intramuscular abscess and bacteremia, and 1 necrotizing fasciitis. Pulsed-field gel electrophoresis revealed that the isolates from 9 patients were clonally related. This clone caused major illness, and 11 of the 13 patients required surgical treatment. Clinicians should be aware of this pathogen and the appropriate broad-spectrum antimicrobial drugs to empirically prescribe for PWID with this life-threatening infection. Close collaboration among public health authorities, outreach social workers, and methadone clinics is essential for timely prevention and control of outbreaks in the PWID population.

Thelaziasis as an uncommon cause of giant papillary conjunctivitis.

INTRODUCTION: We report a local human case of Thelazia callipaeda eye infection in a 49-year-old lady with history of fly contact in Hong Kong. CASE DESCRIPTION: A 49-year-old lady presented with right eye foreign body sensation for one month. She recalled a fly being stuck onto her right upper eyelashes with mascara when she went hiking in a forest trail in Hong Kong. On assessment there were a lot of giant papillae on palpebral conjunctiva. Three living worms crawling on conjunctiva were discovered and removed in total. The worms were identified as Thelazia callipaeda by morphology and molecular sequencing. After removal, her symptoms resolved completely. CONCLUSION: Human thelaziasis is probably under-reported in many countries. The presence of giant papillary conjunctivitis in non-contact lens wearers should alert clinicians to the possibility of thelaziasis in patients with compatible exposure history in endemic regions. Ophthalmologists should increase their awareness towards this uncommon disease and should not wrongly attributed the symptoms to allergic conjunctivitis.

High Prevalence and Frequent Acquisition of Clostridium difficile Ribotype 002 Among Nursing Home Residents in Hong Kong.

OBJECTIVETo determine the incidence and risk factors associated with Clostridium difficile colonization among residents of nursing homes and to identify the ribotypes of circulating C. difficile strains.DESIGNA prospective cohort study with a follow-up duration of 22 months.SETTINGNursing homes.PARTICIPANTSOf the 375 residents in 8 nursing homes, 300 residents (80.0%) participated in the study. A further prospective study of 4 nursing homes involving 141 residents with a minimum of 90 days of follow-up was also performed.METHODSBaseline and 90-day stool cultures were obtained; additional stool cultures were obtained for residents who had been discharged from hospitals. Polymerase chain reaction (PCR) ribotyping and slpA typing were performed for all C. difficile strains isolated.RESULTSToxigenic C. difficile was isolated in 30 residents (10%) at baseline, and 9 residents (7.3%) had acquired toxigenic C. difficile in the nursing homes. The presence of nasogastric tube was an independent risk factor (adjusted odds ratio, 8.59; 95% confidence interval, 1.18-62.53; P=.034) for C. difficile colonization. The Kaplan-Meier estimate of median carriage duration was 13 weeks. The C. difficile ribotypes most commonly identified were 002 (40.8%), 014 (16.9%), 029 (9.9%), and 053 (8.5%).CONCLUSIONSThe high incidence of C. difficile colonization and the overrepresentation of C. difficile ribotype 002 confirmed the contribution of nursing home residents to C. difficile transmission across the continuum of care. An infection control program is needed in long-term care.Infect Control Hosp Epidemiol 2018;782-787.

Clinical features and molecular epidemiology of plasmid-mediated DHA-type AmpC ß-lactamase-producing Klebsiella pneumoniae blood culture isolates, Hong Kong.

Knowledge of risk factors and clinical characteristics of bacteraemia caused by plasmid-mediated AmpC ß-lactamase (pAmpC)-producing Klebsiella pneumoniae (pAmpC-Kp) is not well described. This was a retrospective cohort study of patients with K. pneumoniae bacteraemia in three Hong Kong regional hospitals. Demographic and clinical data were retrieved from medical records. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were performed for molecular epidemiology. A total of 109 patients were included, divided into four groups: bacteraemia due to K. pneumoniae with (i) DHA-type pAmpC (n=23), (ii) extended-spectrum ß-lactamase (ESBL) (n=37), (iii) DHA-type pAmpC+ESBL (n=26) and (iv) controls (n=23). Nursing home residence was independently associated with pAmpC-Kp bacteraemia compared with ESBL-Kp bacteraemia [adjusted odds ratio (aOR)=7.13, 95% confidence interval (CI) 1.36-37.54] and controls (aOR=41.47, 95% CI 4.55-377.75). Compared with controls, patients with pAmpC-Kp bacteraemia also suffered from more severe illness [median Acute Physiology and Chronic Health Evaluation (APACHE) II scores 16 and 25, respectively; P=0.006]. Importantly, the pAmpC group received discordant empirical antimicrobial therapy more frequently (OR=24.00, 95% CI 5.01-114.97), resulting in higher 7-day mortality (OR=20.17, 95% CI 2.32-175.67) and 30-day mortality (OR 4.68, 95% CI 1.29-16.98). PFGE detected six pulsotypes, corresponding to the predominant sequence type 11. Severity of illness and mortality of patients with bacteraemia caused by pAmpC-Kp were high. Patients who are nursing home residents presenting nosocomial sepsis should be treated with broad-spectrum antimicrobials.

Prevalence, prediction, and clonality of methicillin-resistant Staphylococcus aureus carriage at admission to medical units in Hong Kong, China.

OBJECTIVE: To determine the prevalence, risk factors, and molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) colonization at the time of admission to acute medical units and to develop a cost-effective screening strategy. METHODS: Nasal and groin screening cultures were performed for patients at admission to 15 acute medical units in all 7 catchment regions in Hong Kong. All MRSA isolates were subjected to spa typing. RESULTS: The overall carriage rate of MRSA was 14.3% (95% confidence interval [CI], 13.5-15.1). MRSA history within the past 12 months (adjusted odds ratio [OR], 4.60 [95% CI, 3.28-6.44]), old age home residence (adjusted OR, 3.32 [95% CI, 2.78-3.98]), and bed-bound state (adjusted OR, 2.19 [95% CI, 1.75-2.74]) were risk factors selected as MRSA screening criteria that provided reasonable sensitivity (67.4%) and specificity (81.8%), with an affordable burden (25.2%). spa typing showed that 89.5% (848/948) of the isolates were clustered into the 4 spa clonal complexes (CCs): spa CC1081, spa CC032, spa CC002, and spa CC4677. Patients colonized with MRSA spa types t1081 (OR, 1.77 [95% CI, 1.49-2.09]) and t4677 (OR, 3.09 [95% CI, 1.54-6.02]) were more likely to be old age home residents. CONCLUSIONS: MRSA carriage at admission to acute medical units was prevalent in Hong Kong. Our results suggest that targeted screening is a pragmatic approach to increase the detection of the MRSA reservoir. Molecular typing suggests that old age homes are epicenters in amplifying the MRSA burden in acute hospitals. Enhancement of infection control measures in old age homes is important for the control of MRSA in hospitals.