- Volume 68, Issue 5, 2019
Volume 68, Issue 5, 2019
- Review
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Integrons and antibiotic resistance genes in water-borne pathogens: threat detection and risk assessment
More LessAntibiotic-resistant genes (ARGs) are regarded as emerging environmental pollutants and pose a serious health risk to the human population. Integrons are genetic elements that are involved in the spread of ARGs amongst bacterial species. They also act as reservoirs of these resistance traits, further contributing to the development of multi-drug resistance in several water-borne pathogens. Due to inter- and intra-species transfer, integrons are now commonly reported in important water-borne pathogens such as Vibrio , Campylobacter , Salmonella , Shigella , Escherichia coli and other opportunistic pathogens. These pathogens exhibit immense diversity in their resistance gene cassettes. The evolution of multiple novel and complex gene cassettes in integrons further suggests the selection and horizontal transfer of ARGs in multi-drug resistant bacteria. Thus, the detection and characterization of these integrons in water-borne pathogens, especially in epidemic and pandemic strains, is of the utmost importance. It will provide a framework in which health authorities can conduct improved surveillance of antibiotic resistance in our natural water bodies. Such a study will also be helpful in developing better strategies for the containment and cure of infections caused by these bacteria.
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- Antimicrobial Resistance
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Genomic profile of Brazilian methicillin-resistant Staphylococcus aureus resembles clones dispersed worldwide
Purpose. Comparative genomic analysis of strains may help us to better understand the wide diversity of their genetic profiles. The aim of this study was to analyse the genomic features of the resistome and virulome of Brazilian first methicillin-resistant Staphylococcus aureus (MRSA) isolates and their relationship to other Brazilian and international MRSA strains.
Methodology. The whole genomes of three MRSA strains previously isolated in Vitória da Conquista were sequenced, assembled, annotated and compared with other MRSA genomes. A phylogenetic tree was constructed and the pan-genome and accessory and core genomes were constructed. The resistomes and virulomes of all strains were identified.
Results/Key findings. Phylogenetic analysis of all 49 strains indicated different clones showing high similarity. The pan-genome of the analysed strains consisted of 4484 genes, with 31 % comprising the gene portion of the core genome, 47 % comprising the accessory genome and 22 % being singletons. Most strains showed at least one gene related to virulence factors associated with immune system evasion, followed by enterotoxins. The strains showed multiresistance, with the most recurrent genes conferring resistance to beta-lactams, fluoroquinolones, aminoglycosides and macrolides.
Conclusions. Our comparative genomic analysis showed that there is no pattern of virulence gene distribution among the clones analysed in the different regions. The Brazilian strains showed similarity with clones from several continents.
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Synergistic potential of Juniperus communis and Helichrysum italicum essential oils against nontuberculous mycobacteria
Objective. The present study evaluated the possible synergistic antimycobacterial interactions of Juniperus communis and Helichrysum italicum essential oils (EO).
Methods. Antimycobacterial potential was tested against Mycobacterium avium and Mycobacterium intracellulare using broth and water dilution method and checkerboard synergy method. Antiadhesion and antibiofilm effect of EOs was evaluated on biotic (HeLa cells) and abiotic surface (polystyrene). To evaluate the possible mechanisms of action, cellular leakage of proteins and DNA was tested and structural changes were visualized with a transmission electron microscope.
Results. MIC, minimum bactericidal concentration (MBC) and minimal effective concentration (MEC) were 1.6 mg ml−1 for J. communis EO and 3.2 mg ml−1 for H. italicum EO against both mycobacteria. All combinations of EOs in checkerboard synergy method produced fractional inhibitory concentration index values ranging from 0.501 to 1.5, corresponding to synergistic, additive or indifferent effects. Mycobacterium avium showed a greater tendency to create biofilm but these EOs at subinhibitory concentrations (sMIC) effectively blocked the adhesion and the establishment of biofilm. The exposure of both mycobacteria to MICs and sMICs lead to significant morphological changes: acquired a swollen form, ghost-like cell, disorganized cytoplasm detached from the cell wall. OD value of supernatant for both mycobacteria exposed to EOs have confirmed that there is a leakage of cellular material.
Conclusion. The leakage of the cellular material is noticeably higher in sMIC, which is probably due to cell wall damage. sMIC of both EOs have an additive or synergistic effect, reducing MICs, limiting adhesion and preventing the formation of biofilms.
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The potential of fosfomycin for multi-drug resistant sepsis: an analysis of in vitro activity against invasive paediatric Gram-negative bacteria
P urpose . Antimicrobial resistance (AMR) is of increasing global concern, threatening to undermine recent progress in reducing child and neonatal mortality. Repurposing older antimicrobials is a prominent strategy to combat multidrug-resistant sepsis. A potential agent is fosfomycin, however, there is scarce data regarding its in vitro activity and pharmacokinetics in the paediatric population.
M ethodology . We analysed a contemporary, systematically collected archive of community-acquired (CA) and hospital-acquired (HA) paediatric Gram-negative bacteraemia isolates for their susceptibility to fosfomcyin. MICs were determined using agar serial dilution methods and validated by disk diffusion testing where breakpoints are available. Disk diffusion antimicrobial susceptibility testing was also conducted for current empirical therapies (ampicillin, gentamicin, ceftriaxone) and amikacin (proposed in the literature as a new combination empirical therapeutic option).
R esults . Fosfomycin was highly active against invasive Gram-negative isolates, including 90 % (202/224) of Enterobacteriaceae and 96 % (22/23) of Pseudomonas spp. Fosfomycin showed high sensitivity against both CA isolates (94 %, 142/151) and HA isolates (81 %, 78/96; P =0.0015). CA isolates were significantly more likely to be susceptible to fosfomycin than the current first-line empirical therapy (96 % vs 59 %, P <0.0001). Extended spectrum β-lactamases (ESBL) production was detected in 34 % (85/247) of isolates with no significant difference in fosfomycin susceptibility between ESBL-positive or -negative isolates [73/85 (86 %) vs 147/162 (91 %) respectively, P =0.245]. All isolates were susceptible to a fosfomycin-amikacin combination.
C onclusion . Gram-negative paediatric bacteraemia isolates are highly susceptible to fosfomycin, which could be combined with aminoglycosides as a new, carbapenem-sparing regimen to achieve excellent coverage to treat antimicrobial-resistant neonatal and paediatric sepsis.
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- Clinical Microbiology
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Existence of multiple SCCmec elements in clinical isolates of methicillin-resistant Staphylococcus aureus
More LessPurpose. Methicillin-resistant Staphylococcus aureus (MRSA) can be classified into hospital-acquired MRSA (HA-MRSA) and community-associated MRSA (CA-MRSA), based on the associated epidemiological risk factors and their SCCmec types. We therefore studied the diversity and distribution of SCCmec elements among MRSA isolates in our region and also evaluated SCCmec typing as a tool for the classification of MRSA.
Methodology. Two hundred isolates of MRSA obtained from various clinical specimens were included. The clinical and demographic details of the patients and the epidemiological risk factors for MRSA acquisition were documented. Multiplex PCR was optimized for all the major SCCmec types (I to V). Subtyping of SCCmec type IV (IVb, IVc, IVd, IVh) was carried out by simplex PCR.
Results . Based on epidemiological criteria, CA-MRSA constituted 57 % (114/200) of the the test isolates and HA-MRSA made up 43 % (86/200). The predominant SCCmec type found in our study was type III (62%), followed by type V (52.5%) and type I (47.5%), while type II was carried by a single isolate. Of the 200 isolates, 118 carried multiple SCCmec types and 3 were non-typable.
Conclusion. The existence of multiple SCCmec types in individual MRSA isolates resulted in our inability to categorize many of these isolates as either CA-MRSA or HA-MRSA as defined by the SCCmec type criterion.
Limitation . The major limitation of the study was that the SCC mec element of MRSA isolates exhibiting multiple types was not sequenced and hence this finding could not be confirmed.
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Group B streptococcus prevalence, serotype distribution and colonization dynamics in Western Australian pregnant women
More LessPurpose. Streptococcus agalactiae , or group B streptococcus (GBS), is a leading neonatal pathogen that causes sepsis, meningitis and pneumonia. Globally, strategies have been implemented to address vertical transmission, and in Western Australia (WA), culture-based screening at 35–37 weeks’ gestation is part of routine care and guides antibiotic administration. Previous Australian studies have focused on other regions or included low sample-size representatives; we aimed to describe antenatal GBS colonization in WA.
Methodology. A cohort of 814 pregnant women attending antenatal clinics (2015–2017) self-collected vaginal and rectal swabs at ≤22 weeks (n=814) and ≥33 weeks’ (n=567) gestation. These were assessed for GBS presence using culture and PCR, and serotyping was conducted using molecular methods. Lifestyle questionnaires and medical data were collected.
Results. We observed an overall GBS colonization rate of 24%, with 10.6 % of positive participants transiently colonized. Ethnicity (Aboriginal, Torres Strait Islander and African), maternal age ≥25 years, vitamin use, frequent sexual intercourse (≥5 times/week) and use of sex toys were associated with GBS colonization. The dominant serotypes identified were Ia (27.9%), III (20.9%), II (16.3%), V (15.8%), Ib (8.4%), VI (5.1%), IV (2.8%), NT (1.9), VIII (0.5%) and IX (0.5%) at visit one, with V (18.9%) preceding serotype II (18.2%) at visit two. Serotype VII was not detected.
Conclusion. This is the first cohort study to assess GBS colonization in Western Australian pregnant women and will be highly beneficial for guiding clinical practice and future therapeutic options, in particular, the selection of suitable vaccine candidates.
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- Disease, Diagnosis and Diagnostics
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Improvement in serological diagnosis of pertussis by external quality assessment
Purpose. Serological analysis is an essential tool for the diagnosis of pertussis or whooping cough, disease surveillance and the evaluation of vaccine effectiveness against Bordetella pertussis . Accurate measurement of anti-pertussis toxin (anti-PT) IgG antibody levels in sera is essential. These measurements are usually performed using immunological methods such as ELISA and multiplex immunoassays. However, there are a large number of different assay systems available, and therefore standardization and harmonization between the methods are needed to obtain comparable data.
Methodology. In collaboration with ECDC, the EUPert-LabNet network has organized three External Quality Assessment (EQA) schemes (2010, 2012 and 2016), which initially identified the diverse range of techniques and reagents being used throughout Europe. This manuscript discusses the findings of each of the EQA rounds and their impact on the participating laboratories.
Results. The studies have shown an increasing number of laboratories (from 65% to 92%) using only the recommended coating antigen, purified PT, in immunoassays, as this allows exact quantification of serum anti-PT IgG and since PT is only produced by Bordetella pertussis this prevents cross-reactivity with other species. There has also been an increase in the numbers of laboratories (from 59% to 92%), including a WHO reference serum in their assays, which allows anti-PT IgG concentrations to be measured in International Units, thus enabling the comparison of results from different methods and laboratories. In addition, manufacturers have also considered these recommendations when they produce commercial ELISA kits.
Conclusion. The three EQA rounds have resulted in greater harmonization in methods among different laboratories, showing a significant improvement of the ELISA methods used for serodiagnosis of pertussis.
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Clinical evaluation and cost analysis of a Trioplex real-time PCR assay for the detection and differentiation of herpes simplex virus 1 and 2 in cutaneous and mucocutaneous lesions
More LessPurpose. Herpes simplex virus (HSV) is a common lifelong sexually transmitted infection. HSV-1 typically manifests as oral cold sores, while HSV-2 is more traditionally associated with sexual transmission and infection. We have developed a real-time PCR (Trioplex) for the simultaneous detection of HSV-1 and -2 and the bacterial phage internal control (IC) MS2.
Methodology. To determine the performance of the Trioplex method and resolve discrepancies, 178 clinical specimens from cutaneous and mucocutaneous sources were tested using 3 different methods; virus culture with direct fluorescent antibody (DFA) immunostaining, Trioplex and a commercially available HSV analyte-specific reagent (ASR).
Results. HSV Trioplex was significantly more sensitive than virus culture (89 and 67 % HSV 1/2, respectively) and comparable to the commercial assay (P<0.001). Cost analysis revealed that the Trioplex reduced cost by 80 % compared to cell culture.
Conclusions. The implementation of the HSV Trioplex improved the detection turnaround time from 3–10 days to 2.5 h, thus streamlining Herpes detection, improving sensitivity and reducing laboratory costs.
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Inadequate diagnostics: the case to move beyond the bacilli for detection of meningitis due to Mycobacterium tuberculosis
More LessTuberculosis (TB) meningitis is extremely difficult to diagnose due to its pauci-bacillary disease nature and new techniques are needed. Improved test sensitivity would allow for greater clinician confidence in diagnostic testing and has the potential to improve patient outcomes. Traditional microbiologic and molecular tests for TB meningitis focus on detection of TB bacilli and are inadequate. Smear microscopy is rapid but only ~10–15 % sensitive. Culture has 50–60 % sensitivity but is slow. Xpert MTB/Rif Ultra is a rapid, automated PCR-based assay with ~70 % sensitivity versus clinical case definition. Thus, even the best current testing may miss up to 30 % of cases. Clinicians are often left to treat empirically with prolonged regimens with significant side effects or risk a missed case that would result in death. Rather than relying strictly on microbiologic or molecular testing to diagnose TB meningitis, we propose that testing of CSF for biomarkers of host response may have an adjunctive role to play in improving the diagnosis of TB meningitis.
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Molecular detection of Treponema pallidum subspecies pallidum in 150-year-old foetal remains, southeastern France
More LessPurpose. Syphilis, caused by Treponema pallidum subspecies pallidum , is considered as an old disease affecting humans; traces of such infections, including congenital syphilis, are potentially identifiable in archaeological samples. The aim of this research was to perform macroscopic and molecular investigations of T. pallidum on six infant remains, buried between 1837 and 1867, from the cemetery of ‘Les Crottes’ in Marseille city (southeastern France).
Methodology. Pathological analysis of bones from individuals, aged from the twenty-ninth week of amenorrhea to 4–9 months, was performed. Samples served also as a source of ancient DNA (aDNA) for PCR-based molecular investigations targeting T. pallidum DNA; all samples were also tested for Mycobacterium tuberculosis and Plasmodium falciparum DNA. Sequences characterized were cloned and sequenced, and compared to those available in databases.
Results/Key findings. All samples tested displayed widespread osteoporotic lesions across the skeleton possibly related to some metabolic or infectious disorders. Subsequent molecular analysis revealed that one individual, SP332 (unborn, 29 amenorrhea weeks, inhumation date 1864–1866), exhibited positive signals for the five T. pallidum amplification systems tested; sequence analysis provided strong evidence for the effective detection of T. pallidum subspecies pallidum DNA.
Conclusions. Individual SP332 is the first PCR-confirmed palaeopathological case of syphilis identified in France, and the youngest specimen ever to be diagnosed with certainty for congenital syphilis. Future research aimed at better characterizing this 150-year-old treponeme genome and exploring new archaelogical cases of syphilis in the very young should contribute to a better comprehension of the disease's history.
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- Medical Mycology
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Molecular characterization and antifungal susceptibility testing of Candida nivariensis from blood samples – an Iranian multicentre study and a review of the literature
Purpose. Identification of the emerging yeast species Candida nivariensis among presumptively identified Iranian Candida glabrata isolates.
Methodology. Clinical C. glabrata species complex isolates from blood (n=100; 46.9%), vaginal swabs (n=20; 9.4%), bronchoalveolar lavage (n=10; 4.7%) and sputum (n=12; 5.6%) from 68 patients from Iran were investigated. Isolates were characterized by CHROMagar, multiplex PCRs, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), amplified fragment length polymorphism (AFLP) fingerprinting, internal transcribed spacer (ITS)/large subunit (LSU) rDNA and FKS1/FKS2 sequencing, and the European Committee on Antimicrobial Susceptibility Testing broth microdilution method. A comprehensive literature review was conducted and all the relevant clinical and microbiological data were collected.
Results. Four C. nivariensis isolates were recovered from blood samples of three subjects and were all consistently identified by nine-plex PCR, Bruker MALDI-TOF MS, and LSU and ITS rDNA sequencing. AFLP genotyping clustered the isolates into two groups. Sequencing of the FKS1 and FKS2 hotspots showed no accountable amino acid substitutions. All isolates were susceptible to amphotericin B, fluconazole, itraconazole, posaconazole, voriconazole, anidulafungin and micafungin.
Conclusion. In total, 4 out of 213 clinical C. glabrata species complex candidemia isolates were C. nivariensis. Improvement of the BioMerieux Vitek MS database is required to accurately identify C. nivariensis and it is advised to alternatively use CHROMagar and/or PCR-based techniques. As other species within the Nakaseomyces clade may cause infection and showed high MIC values for antifungals, inclusion of their spectra into the MALDI-TOF MS database seems relevant. Due to developing resistance to fluconazole and insufficient efficacy of caspofungin, the combination of catheter removal plus treatment with caspofungin, or voriconazole, or micafungin might be effective for patients.
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Epidemiology and susceptibility profile to classic antifungals and over-the-counter products of Malassezia clinical isolates from a Portuguese University Hospital: a prospective study
Purpose. Clinical epidemiological data about the distinct Malassezia species remain scarce. The recurrence of Malassezia-related skin diseases, despite long-term use of antifungals, raises concern about the hypothetical emergence of antifungal resistance. We aimed to assess the distribution of Malassezia species among patients from a University Hospital with pityriasis versicolor, seborrheic dermatitis and healthy volunteers, and to evaluate the susceptibility profile to classic antifungals and over-the-counter compounds, searching for clinical associations.
Methodology. The enrollment of volunteers was conducted at the Dermatology Department of a University Hospital over a 3 year period. Malassezia culture isolates were identified to the species-level by sequencing. The drug susceptibility profile was assessed according to a broth microdilution assay, as recommended by the Clinical Laboratory Standards Institute.
Results. A total of 86 Malassezia isolates were recovered from 182 volunteers. Malassezia sympodialis was the most frequent isolated species. We found high MIC values and a wide MIC range in the case of tested azoles, and very low terbinafine MIC values against most isolates. Previous topical corticosteroid therapy was associated with a significant increase of MIC values of fluconazole and of terbinafine.
Conclusion. Conversely to other European studies, M. sympodialis was the most common isolated species, which might be related to geographic reasons. The impact of previous topical corticotherapy upon the antifungal susceptibility profile was hereby demonstrated. In vitro susceptibility test results suggest that terbinafine might be a valid alternative for Malassezia-related skin diseases nonresponsive to azoles.
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- Molecular and Microbial Epidemiology
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Polyclonal spread of multiple genotypes of Mycoplasma pneumoniae in semi-closed settings in Yamagata, Japan
Purpose. To clarify the spread of Mycoplasma pneumoniae infections in semi-closed settings such as schools and family homes using molecular typing methods.
Methodology. We retrospectively searched for school- and family-based clusters of M. pneumoniae infections based on information regarding patients from whom M. pneumoniae strains had been isolated between 2011 and 2013 in Yamagata, Japan. The molecular typing profile, including the P1 type and the four-locus (Mpn13, 14, 15 and 16) multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) type, was obtained from our previous study.
Results. We identified 11 school-based clusters involving 71 patients and 16 family-based clusters involving 38 patients, including 14 duplications between these types of clusters. A total of 95 M. pneumoniae strains isolated from those patients were divided into 4 genotypes: 33 strains of type 4-5-7-2, 1; 31 of type 4-5-7-3, 1; 24 of type 3-5-6-2, 2c; and 7 of type 3-5-6-2, 2a. Of the 11 school-based clusters, 6 clusters (54.5%) consisted of multiple genotypes, and the remaining 5 clusters consisted of a single genotype. Moreover, the presence of multiple genotypes was identified in three classrooms of a school. On the other hand, in 14 (87.5%) of the 16 family-based clusters, the genotypes of the M. pneumoniae strains isolated from each family member were identical.
Conclusion. The spread of M. pneumoniae infection in schools is likely polyclonal, since M. pneumoniae strains are brought into schools from various sites, such as family homes, which are important sites of disease transmission.
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- Prevention, Therapy and Therapeutics
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Synthesis of conjugated PIA–rSesC and immunological evaluation against biofilm-forming Staphylococcus epidermidis
Purpose. Staphylococcus epidermidis is an opportunistic pathogen and a leading cause of morbidity and premature mortality in patients with medical device-related infections, which is a concern in hospitalized patients. Developing a strategy to raise opsonic antibodies against polysaccharide intracellular adhesin (PIA) could be promising for the elimination of colonizing and biofilm-forming S. epidermidis . Following the purification of truncated rSesC protein and PIA, for the first time, PIA was conjugated to rSesC as a carrier to increase the immunogenicity of PIA and its efficacy in mice was evaluated. The structure of the conjugate was analysed using the Fourier transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance spectroscopy (H1- NMR) methods. Afterwards, the immune response was evaluated by measuring the total IgG, IgG2a and IgG2b titres.
Results. The immunization of mice with the PIA–rSesC conjugate raised the levels of opsonic antibodies, and the vaccinated mice were protected when challenged intravenously by wild-type S. epidermidis strain 1457. Further studies indicated that the conjugated vaccine was able to eliminate S. epidermidis biofilm formation in in vitro or in vivo assays.
Conclusion. This study confirms the proposal that the immunization of mice with PIA-rSesC conjugate vaccine could protect against S. epidermidis infection.
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Outcome of a screening program for the prevention of neonatal early-onset group B Streptococcus infection: a population-based cohort study in Inner Mongolia, China
More LessPurpose . Invasive early-onset group B Streptococcus infection (EOGBS) is an important cause of severe neonatal complications but study on comprehensive GBS screening is lacking in China. This study aims to investigate the outcome of a regional anterpartum screening program for EOGBS prevention and to estimate the pros and cons of a new GBS screening strategy employed.
Methods . We performed an optimized hospital strategy for GBS screening, which targeted expectant mothers (including those with preterm births) from January 2016 to December 2016 in a population-based cohort. Three common screening strategies were simulated to estimate the availability of the hospital strategy used in this study.
Results . Altogether, 9770 eligible women were tested and the rate of GBS carriage was 2.7 % (266/9770). In total, 198 of the 266 maternal GBS carriers accepted intrapartum antibiotic prophylaxis (IAP) treatment. Among the 9860 neonates of 9770 mothers, four cases of EOGBS infection were identified and one case was missed (EOGBS incidence with screening and IAP: 0.5/1000). Risk factors for maternal GBS colonization included preterm birth (between 35 and 37 weeks) [odds ratio (OR)=1.7 (95 % confidence interval: 1.22–2.33)], region of origin, resident areas, maternal age (older than 34 years) [OR=1.5 (1.06–2.09)], prelabour rupture of membranes [OR=1.8 (1.34–2.35)], gestational diabetes mellitus [OR=1.6 (1.14–2.28)] and maternal mild anemia (Hb: 90–110 g dl−1) [OR=1.5 (1.16–2.06)]. This new screening strategy resulted in less antibiotic exposure and least number of cases missed.
Conclusions . Our findings illustrate that this perinatal screening (including preterm births) for prevention of EOGBS infection can be implemented in the Inner Mongolian area.
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- Method
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Identifying indoor air Penicillium species: a challenge for allergic patients
More LessPurpose. Penicillium is the most common mould isolated in housing. Penicillium chrysogenum is the only species tested by prick test or serology for allergic patients. The American Institute of Medicine has accepted Penicillium as an aetiological agent of rhinitis in children and adults and as an asthma agent in children. However, few studies have identified Penicillium in housing to the species level (354 species). Phenotypic identification is difficult. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) should be an alternative. The aim of this study was (1) to identify the Penicillium species present in dwellings in Eastern France and (2) to evaluate the reliability of MALDI-TOF MS for identification, by comparing it to DNA sequencing and phenotypic identification.
Methodology. Identification to the species level was performed by MALDI-TOF MS on 275 strains isolated from 48 dwellings. These results were compared to beta-tubulin gene sequencing and to the phenotypic aspects.
Results. Thanks to MALDI-TOF, 235/275 strains could be identified (85.5 %). Fourteen species were identified among 23 Penicillium species included in the Filamentous Fungi Library 1.0 (Bruker Daltonics). However, 72.2 % of the strains belonged to five main taxa: P. chrysogenum (27.3 %), Penicillium glabrum (22.9 %), Penicillium commune (11.3 %), Penicillium brevicompactum (6.5 %) and Penicillium expansum (4.2 %).
Conclusion. Complete coherence between MALDI-TOF MS and sequence-based identification was found for P. chrysogenum, P. expansum, P. glabrum, Penicillium italicum and Penicillium corylophilum. The main drawback was observed for Penicillium crustosum, which included 21 strains (7.6 %) that could not be identified using MALDI-TOF MS.
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Volumes and issues
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Volume 73 (2024)
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Volume 72 (2023 - 2024)
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