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147500.9911Evaluation of the FilmArray(®) Pneumonia Plus Panel for Rapid Diagnosis of Hospital-Acquired Pneumonia in Intensive Care Unit Patients. The FilmArray(®) Pneumonia plus Panel (FAPP) is a new multiplex molecular test for hospital-acquired pneumonia (HAP), which can rapidly detect 18 bacteria, 9 viruses, and 7 resistance genes. We aimed to compare the diagnosis performance of FAPP with conventional testing in 100 intensive care unit (ICU) patients who required mechanical ventilation, with clinically suspected HAP. A total of 237 samples [76 bronchoalveolar lavages (BAL(DS)) and 82 endotracheal aspirates (ETA(DS)) obtained at HAP diagnosis, and 79 ETA obtained during follow-up (ETA(TT))], were analyzed independently by routine microbiology testing and FAPP. 58 patients had paired BAL(DS) and ETA(DS). The positivity thresholds of semi-quantified bacteria were 10(3)-10(4) CFUs/mL or 10(4) copies/mL for BAL, and 10(5) CFUs/mL or copies/mL for ETA. Respiratory commensals (H. influenzae, S. aureus, E. coli, S. pneumoniae) were the most common pathogens. Discordant results for bacterial identification were observed in 33/76 (43.4%) BAL(DS) and 36/82 (43.9%) ETA(DS), and in most cases, FAPP identified one supplemental bacteria (23/33 BAL(DS) and 21/36 ETA(DS)). An absence of growth, or polybacterial cultures, explained almost equally the majority of the non-detections in culture. No linear relationship was observed between bin and CFUs/mL variables. Concordant results between paired BAL(DS) and ETA(DS) were obtained in 46/58 (79.3%) patients with FAPP. One of the 17 resistance genes detected with FAPP (mecA/C and MREJ) was not confirmed by conventional testing. Overall, FAPP enhanced the positivity rate of diagnostic testing, with increased recognition of coinfections. Implementing this strategy may allow clinicians to make more timely and informed decisions.202032983057
125410.9911Genetic diversity and antimicrobial resistance of Staphylococcus aureus from recurrent tonsillitis in children. The aim of this study was to analyze the prevalence of Staphylococcus aureus in the tonsils of children subjected tonsillectomy due to recurrent tonsilitis and to determine the spa types of the pathogens, carriage of virulence genes and antimicrobial resistance profiles. The study included 73 tonsillectomized children. Bacteria, including S. aureus were isolated from tonsillar surface prior to tonsillectomy, recovered from tonsillar core at the time of the surgery, and from posterior pharynx 2-4 weeks after the procedure. Staphylococcus aureus isolates were compared by spa typing, tested for antimicrobial susceptibility and for the presence of superantigenic toxin genes (sea-seu, eta, etb, tst, lukS/lukF-PV) by multiplex polymerase chain reaction. Seventy-three patients (mean 7.1 ± 4.1 years, 61.6% male) were assessed. The most commonly isolated bacteria were S. aureus. The largest proportion of staphylococcal isolates originated from tonsillar core (63%), followed by tonsillar surface (45.1%) and posterior pharynx in tonsillectomized children (18.2%, p = 0.007). Five (6.3%) isolates were identified as MRSA (mecA-positive). Up to 67.5% of the isolates synthesized penicillinases (blaZ-positive isolates), and 8.8% displayed MLS(B) resistance. The superantigenic toxin genes were detected in more than half of examined isolates (56.3%). spa types t091, t084, and t002, and clonal complexes (CCs) CC7, CC45, and CC30 turned out to be most common. Staphylococcus aureus associated with RT in children showed pathogenicity potential and considerable genetic diversity, and no clones were found to be specific for this condition although further studies are needed.202031692060
218920.9909High prevalence of Panton-Valentine Leucocidin (PVL) toxin carrying MRSA and multidrug resistant gram negative bacteria in late onset neonatal sepsis indicate nosocomial spread in a Pakistani tertiary care hospital. BACKGROUND: Neonatal sepsis has high incidence with significant mortality and morbidity rates in Pakistan. We investigated common etiological patterns of neonatal sepsis at a tertiary care setup. METHODS: 90 pus and blood, gram negative and gram positive bacterial isolates were analyzed for virulence and antibiotic resistance gene profiling using PCR and disc diffusion methods. RESULTS: Staphylococcus aureus showed strong association with neonatal sepsis (43 %) followed by Citrobacter freundii (21 %), Pseudomonas aeruginosa (13 %), Escherichia coli (15 %) and Salmonella enterica (8 %). Molecular typing of E. coli isolates depicted high prevalence of the virulent F and B2 phylogroups, with 4 hypervirulent phylogroup G isolates. 76.9 % S. aureus isolates showed presence of Luk-PV, encoding for Panton-valentine leucocidin (PVL) toxin with majority also carrying MecA gene and classified as methicillin resistant S. aureus (MRSA). ecpA, papC, fimH and traT virulence genes were detected in E. coli and Salmonella isolates. 47 % Citrobacter freundii isolates carried the shiga like toxin SltII B. Antimicrobial resistance profiling depicted common resistance to cephalosporins, beta lactams and fluoroquinolones. CONCLUSION: Presence of PVL carrying MRSA and multidrug resistant gram negative bacteria, all isolated from late onset sepsis neonates indicate a predominant nosocomial transmission pattern which may complicate management of the disease in NICU setups.202336621204
217030.9908Drug resistance in bacteria isolated from patients presenting with wounds at a non-profit Surgical Center in Phnom Penh, Cambodia from 2011-2013. BACKGROUND: Emerging antibiotic resistance amongst clinically significant bacteria is a public health issue of increasing significance worldwide, but it is relatively uncharacterized in Cambodia. In this study we performed standard bacterial cultures on samples from wounds at a Non-Governmental-Organization (NGO) Hospital in Phnom Penh, Cambodia. Testing was performed to elucidate pathogenic bacteria causing wound infections and the antibiotic resistance profiles of bacterial isolates. All testing was performed at the Naval Medical Research Unit, No.2 (NAMRU-2) main laboratory in Phnom Penh, Cambodia. METHODS: Between 2011-2013, a total of 251 specimens were collected from patients at the NGO hospital and analyzed for bacterial infection by standard bacterial cultures techniques. Specimens were all from wounds and anonymous. No specific clinical information accompanied the submitted specimens. Antibiotic susceptibility testing, and phenotypic testing for extended-spectrum beta-lactamase (ESBL) were performed and reported based on CLSI guidelines. Further genetic testing for CTX-M, TEM and SHV ESBLs was accomplished using PCR. RESULTS: One-hundred and seventy-six specimens were positive following bacterial culture (70 %). Staphlycoccus aureus was the most frequently isolated bacteria. Antibiotic drug resistance testing revealed that 52.5 % of Staphlycoccus aureus isolates were oxacillin resistant. For Escherichia coli isolates, 63.9 % were ciprofloxacin and levofloxacin resistant and 96 % were ESBL producers. Resistance to meropenem and imipenem was observed in one of three Acinetobacter spp isolates. CONCLUSIONS: This study is the first of its kind detailing the antibiotic resistance profiles of pathogenic bacteria causing wound infections at a single surgical hospital in Cambodia. The reported findings of this study demonstrate significant antibiotic resistance in bacteria from injured patients and should serve to guide treatment modalities in Cambodia.201528883936
148140.9908Molecular versus conventional assay for diagnosis of hospital-acquired pneumonia in critically ill patients: a single center experience. PURPOSE: Lower respiratory tract infections are reported as one of top five causes of mortality and morbidity in the world. A bacterial etiology is often involved in HAP, most frequently from multidrug resistant gram-negative bacteria, and fast accurate diagnosis of etiologic agent(s) of LRTI is essential for an appropriate management. The aim of this retrospective study was to evaluate the analytical performance of Biofire Filmarray Pneumonia Plus for bacteria detection in bronchoalveolar lavage samples and the concordance of bacterial loads between BFPP and cultural gold standard methods. METHODS: A total of 111 BAL samples were obtained from 111 consecutive patients admitted to Intensive Care Unit of "Renato Dulbecco" Teaching Hospital of Catanzaro, from March 2023 to March 2024. RESULTS: Compared to conventional methods, BFPP showed a sensitivity of 99 % and a specificity of 64 %. The agreement between the two methods was assessed by calculating PPA and NPA, being 89 % and 95 %, respectively. The most common bacterial species identified at BFPP was Klebsiella pneumoniae, followed by Acinetobacter calcaceuticus-baumanii complex, Staphylococcus aureus and Pseudomonas aeruginosa. Bacterial load (CFU/ml) in relation to copy number detected by molecular analysis showed the best performance for value ≥10(6) copie/mL. About molecular mechanisms of resistance in comparison to phenotypic profiles, the highest level of performance was observed for presence of KPC genes, all isolates showing resistance to carbapenems, followed by OXA-48 like and NDM. CONCLUSION: The high concordance reported in this study between the identification of resistance genes and phenotypic indication can lead to an appropriate, fast and tailored antibiotic therapy.202540513663
234450.9907Analysis of antibiotic resistance and genetic profile of conjunctival bacteria flora before and after cataract surgery. PURPOSE: To analyze antibiotic resistance and genetic profile of conjunctival bacteria flora before and after cataract surgery with the focus on coagulase-negative staphylococci (CNS) during cataract surgery and discuss the implications of this colonization as a potential risk of acquiring endophthalmitis. METHODS: After approval of the institutional review board and informed consent from patients had been obtained, conjunctival swabs for culture from 59 patients undergoing cataract surgery were taken of the fellow eye at baseline (C0) and from the eye to be operated before (T0) and after (T1) irrigation with povine-iodine 5%, and at the end of surgery (T2). Genes responsible for virulence (mecA, ica and atlE) and antibiotic profile were determined; strain clonality of persistent colonizing Staphylococcus epidermidis strains was established by the Multi-locus sequence typing (MLST). RESULTS: The frequency of CNS was significantly reduced in T1 (13.6%) from 81.4% in T0 and 86.4% in C0. The frequency of mecA, ica and atlE genes was 34.4%, 37.5% and 61.4%, respectively; and methicillin phenotypic resistance was 35.4%. S. epidermidis was the most frequent species isolated in every time point. MLST revealed in 7 patients 100% coincidence of the seven alleles of the S. epidermidis isolated previous to povine-iodine 5% disinfection and at the end of the surgery. CNS isolates from T1 or T2 corresponded to the same species, antibiotic and virulence profile as those isolates from C0 or T0. CONCLUSION: Povidone-iodine 5% prophylaxis before surgery significantly reduced conjunctival contamination; in those that persisted, the source of contamination was mostly the patient's microbiota confirmed by the MLST system.202335943639
145560.9907Resistance to bacterial infection, complication occurring after cardiac surgery. To analyze the occurrence of resistant bacterial infection in patients undergoing cardiac surgery hospitalized in the surgical specialty hospital, in Erbil city, Iraq. A prospective study was done on a total of 138 patients operated and hospitalized in an intensive care unit and surgical wards. Bacterial isolates identification was done according to cultural characteristics, microscopic examination, some biochemical tests, analytic Profile Index 20E& API Staph, confirmed with VITEK® 2 compact system (BioMérieux). Antimicrobial susceptibility for disc diffusion tested to 17 antimicrobial agents. Resistance isolates were confirmed phenotypically for carbapenemase by Rapidec Carba NP Test (bioMe´rieux SA, Marcy-l'E´toile, France) for ESBLs producers by ESBL screening test VITEK 2 system. Molecularly blaIMP blaTEM, blaKPC, AmpC and blaCTX-M were detected by PCR. In 134 patients, 28.3% of patients got infected post-operatively. The most frequent source of isolation was from ICU patients (75%). Isolated bacteria included gram-positive 29 (54.7%) and gram-negative bacteria 24 (45.3%). Most frequently:  Staphylococcus aureus (24.4%), each of pseudomonas aeroginosa, Klebsiella pneumonia (15.1%), Streptococcus spp. (11.3%), Escherichia coli (9.4%). Whereas included Coagulase Negative Staphylococci species (CoNS) (13.2%) and Enterococci species (5.7) Statistical analysis showed significantly higher sensitive isolates as compared with resistance isolates. Resistance to Carbapenems calss was 18.9% and Cephalosporins class 41.5% of isolates. The antimicrobial resistance pattern indicated that MDR bacterial isolates (81.1%) were widespread. Of the 34 phenotypically ESBL positive isolates, the ESBL genes (AmpC, blaCTX-M, and blaTEM) were amplified in 7(20.6), 6(17.6) and 6(17.6) isolates respectively. Out of 8 K. pneumonia (37.5%) harboring both blaAmpC and bla-CTX-M genes, while 6(75%) carries blaTEM. The blaCTX-M gene was found in only 1 (12.5%) out of 8 isolates of P. aeruginosa. While blaAmpC genotyping revealed that 1(7.7%) out of 13 Staph. aureus isolates were harboring it. Finally, 3(60%) out of 5 E. coli isolates harboring both AmpC and bla-CTX-M genes. Cardiac surgery patients wound show increasingly emerging strains of ESBL-producing gram-negative bacteria K. pneumonia, P. aeruginosa and E. coli especially patients prolonged in the intensive care unit.202034174972
125770.9907Antimicrobial Susceptibility Pattern in the Bacteria Isolated from Surgical Site Infection: Emphasis on Staphylococcus Aureus; Yasuj City, Southwest Iran. BACKGROUND: Surgical site infections (SSIs) in surgical wards remains the most common cause of postoperative complications and realistically is the third most common origin of healthcare-related conditions. Staphylococcus aureus is undoubtedly the most common bacteria causing SSIs. The current study aimed at investigating the antimicrobial susceptibility pattern in bacteria isolated from SSIs, evaluation of tetracycline resistance genes, and SCCmec typing in S. aureus isolates isolated from patients with SSIs from 2018 to 2019 in Yasuj, Kohgiluyeh, and Boyer-Ahmad Province, Iran. METHODS: This study diligently investigated 240 potential patients. Antimicrobial susceptibility testing was performed properly by the disk diffusion method. For the final confirmation of isolated bacteria, PCR was used. The presence of tet genes and SCCmec typing was carried out by multiplex PCR. RESULTS: The results showed that the most common isolated pathogens included S. aureus, E. coli, P. aeruginosa, Coagulase-negative Staphylococci, and K. pneumonia in 58.8%, 19.8%, 9.2%, 6.8% and 5.4% of cases, respectively. The majority of the Gram positive isolates were resistant against penicillin (86%) and Gram negative were resistant against ciprofloxacin (75.6%). In isolates of Staphylococcus aureus, the mecA gene was detected in 63.6% of isolates. The predominant SCCmec types were type III (59.1%) and type I (18.4%). The tetK and tetM genes were detected in 80.7% and 71.9% of the S. aureus isolates, respectively. There was a statistically significant difference between tet genes (tetK and tetM) from the viewpoint of resistance to tetracycline (p = 0.024). CONCLUSIONS: According to the results of the current study, it is recommended to administer vancomycin, amikacin, and imipenem in Yasuj to treat SSIs.202133616327
145480.9906OCCURRENCE OF AMINOGLYCOSIDES RESISTANCE GENES ACC(6)-IB AND ACC(3)-II AMONG GRAM-NEGATIVE ISOLATES CAUSING URINARY TRACT INFECTION IN PEDIATRIC PATIENTS, NAJAF, IRAQ. OBJECTIVE: The aim: The aim of the study was to detect the antimicrobial susceptibility patterns and frequency of aminoglycosides resistance genes of Gram-negative bacteria isolated from pediatric patient with UTI. PATIENTS AND METHODS: Materials and methods: The study has been performed with a total of 500 urine specimens collected from pediatric patients under the age of 18 year suspected with UTI, admitted to hospitals in Al-Najaf province/Iraq during the period from November 2018 to March 2019. RESULTS: Results: A total of 500 urine specimens had been tested, 120 (24%) had signifficant bacteriuria, while there 380 (76%) had non-signi!cant bacteriuria. Escherichia coli represent about 70 (68.2%) followed by followed by 23 (22.5%) K. pneumoniae, 5 (4.9%) P. aeruginosa, 2 (1.9%) Proteus spp., 1 (0.9%) Enterobacter spp. and 1 (0.9%) Oligella uratolytic. The antimicrobial susceptibility profile of 102 Gram-negative isolates, revealed that 59 (58%) were multidrug resistant (MDR) and 38(37%) were extensive drug resistant (XDR). The PCR results of aminoglycosides resistance showing that 23 (74.1%) Gram-negative isolates had acc(6')-Ib gene and 12 (38.7%) Gram-negative isolates acc(3')-II gene. CONCLUSION: Conclusions: A high frequency of multi-drug resistance and extensive-drug resistance of isolates were recognized, and an alarming percentage of amino-glycosides resistance to acc(6')-Ib and acc(3')-II.202337010165
147390.9906Evaluation of the Unyvero i60 ITI® multiplex PCR for infected chronic leg ulcers diagnosis. OBJECTIVES: Unyvero i60 ITI multiplex PCR (mPCR) may identify a large panel of bacteria and antibiotic resistance genes. In this study, we compared results obtained by mPCR to standard bacteriology in chronic leg ulcer (CLU) infections. METHODS: A prospective study, part of the interventional-blinded randomized study "ulcerinfecte" (NCT02889926), was conducted at Saint Joseph Hospital in Paris. Fifty patients with a suspicion of infected CLU were included between February 2017 and September 2018. Conventional bacteriology and mPCR were performed simultaneously on deep skin biopsies. RESULTS: Staphylococcus aureus and Pseudomonas aeruginosa were the most detected pathogens. Regarding the global sensitivity, mPCR is not overcome to the standard culture. Anaerobes and slow growing bacteria were detected with a higher sensitivity rate by mPCR than standard culture. CONCLUSION: Unyvero i60 ITI multiplex PCR detected rapidly pathogenic bacteria in infected CLU especially anaerobes and slow growing bacteria and was particularly effective for patients previously treated with antibiotics.202031790779
2188100.9904Detection of Virulence Factors and Antibiotic Resistance Pattern of Clinical and Intensive Care Unit Environmental Isolates of Pseudomonas aeruginosa. BACKGROUND: Pseudomonas aeruginosa is a gram-negative non-glucose fermenting aerobic bacteria and an opportunistic pathogen in humans and animals. The present study was carried out to investigate the distribution of virulence factors and antibiotic resistance properties of P. aeruginosa isolated from patients and intensive care unit (ICU) environment. MATERIAL AND METHODS: A total of 116 P. aeruginosa isolated from patients and ICU environment were collected from Besat hospital in Hamadan, the West of Iran. P. aeruginosa isolates were analyzed based on the presence of the virulence factors encoding genes included exoA, exoS, exoU, and algD using polymerase chain reaction (PCR). Antimicrobial susceptibility test was performed using a disk diffusion method. RESULTS: The results showed the prevalence of exoA 33 (56.9%), exoS 21 (36.20%), exoU 37 (63.8%), and algD 35 (60.34%) genes in ICU environment P. aeruginosa strains and exo A 23 (39.25%), exoS 25 (43.1%), exoU 40(68.98%), and algD 25 (43.1%) genes in clinical isolates of P. aeruginosa. High resistance levels of the clinical and ICU environment isolate to ampicillinsulbactam (100%), were also observed. CONCLUSION: Our findings should raise awareness about antibiotic resistance in hospitalized patients in Iran. Clinicians should exercise caution in prescribing antibiotics, especially in cases of human infections.202031889501
1406110.9904Multicentre study of the burden of multidrug-resistant bacteria in the aetiology of infected diabetic foot ulcers. BACKGROUND: Infected diabetic foot ulcer (IDFU) is a public health issue and the leading cause of non-traumatic limb amputation. Very few published data on IDFU exist in most West African countries. OBJECTIVE: The study investigated the aetiology and antibacterial drug resistance burden of IDFU in tertiary hospitals in Osun state, Nigeria, between July 2016 and April 2017. METHODS: Isolates were cultured from tissue biopsies or aspirates collected from patients with IDFU. Bacterial identification, antibiotic susceptibility testing and phenotypic detection of extended-spectrum beta-lactamase and carbapenemase production were done by established protocols. Specific resistance genes were detected by polymerase chain reaction. RESULTS: There were 218 microorganisms isolated from 93 IDFUs, comprising 129 (59.2%) Gram-negative bacilli (GNB), 59 (27.1%) Gram-positive cocci and 29 (13.3%) anaerobic bacteria. The top five facultative anaerobic bacteria isolated were: Staphylococcus aureus (34; 15.6%), Escherichia coli (23; 10.6%), Pseudomonas aeruginosa (20; 9.2%), Klebsiella pneumoniae (19; 8.7%) and Citrobacter spp. (19; 8.7%). The most common anaerobes were Bacteroides spp. (7; 3.2%) and Peptostreptococcus anaerobius (6; 2.8%). Seventy-four IDFUs (80%) were infected by multidrug-resistant bacteria, predominantly methicillin-resistant S. aureus and GNB producing extended-spectrum β-lactamases, mainly of the CTX-M variety. Only 4 (3.1%) GNB produced carbapenemases encoded predominantly by bla (VIM). Factors associated with presence of multidrug-resistant bacteria were peripheral neuropathy (adjusted odds ratio [AOR] = 4.05, p = 0.04) and duration of foot infection of more than 1 month (AOR = 7.63, p = 0.02). CONCLUSION: Multidrug-resistant facultative anaerobic bacteria are overrepresented as agents of IDFU. A relatively low proportion of the aetiological agents were anaerobic bacteria.202133824857
1479120.9904BioFire FilmArray BCID2 versus VITEK-2 System in Determining Microbial Etiology and Antibiotic-Resistant Genes of Pathogens Recovered from Central Line-Associated Bloodstream Infections. Central line-associated bloodstream infection (CLABSI) is among the most serious hospital acquired infections. Therefore, the rapid detection of the causative microorganism is of crucial importance to allow for the appropriate antimicrobial therapy. In the present study, we analyzed the clinical performance of the BioFire FilmArray Blood Culture Identification 2 (BCID2) panel in the identification of 33 microbial species and 10 antibiotic resistance genes in comparison to the VITEK-2 system. A total of 104 blood specimens were included. The FilmArray BCID2 results were concordant with the VITEK-2 system in 69/97 specimens (71.1%). Non-concordance was either due to the detection of more pathogens by the FilmArray BCID2 23/28 (82%) or microbial species were misidentified 5/28 (18%). Hence, in comparison to the VITEK-2 system, the FilmArray BCID2 panel showed an overall sensitivity of 75.8% (95% CI, 66-83%) and an overall specificity of 98% (95% CI, 97-98.8%) in detecting microbial species. For the resistance genes, the FilmArray BCID was able to detect the presence of blaCTX-M gene in 23 Gram-negative isolates, blaNDM and blaOXA-48- like genes in 14 and 13 isolates, respectively. The mecA and mecC genes were found in 23 Staphylococcus species, while mecA, mecC and MREJ genes were found in 4 Staphylococcus aureus isolates. The sensitivity and specificity for detecting resistance genes by the FilmArray BCID2 was 90% (95% CI, 81.4-95%) and 99.6% (95% CI, 99-100%), respectively. As concluded, the present study emphasizes the high sensitivity and specificity of the FilmArray BCID2 in the rapid and reliable detection of different bacteria and fungi from positive blood culture bottles, as well as the accurate detection of various antibiotic resistance markers.202236358274
1428130.9904Carbapenem-resistant Gram-negative bacteria associated with catheter-related bloodstream infections in three intensive care units in Egypt. We aimed to identify the carbapenem-resistant Gram-negative bacteria (GNB) causing catheter-related bloodstream infections (CRBSI) in intensive care units (ICU) in a tertiary care Egyptian hospital, to study their resistance mechanisms by phenotypic and genetic tests, and to use ERIC-PCR for assessing their relatedness. The study was conducted over 2 years in three ICUs in a tertiary care hospital in Egypt during 2015-2016. We identified 194 bloodstream infections (BSIs); 130 (67.01%) were caused by GNB, of which 57 were isolated from CRBSI patients (73.84%). Identification of isolates was performed using conventional methods and MALDI-TOF MS. Antimicrobial susceptibility testing (AST) was done by disc diffusion following CLSI guidelines. Phenotypic detection of carbapenemases enzymes activity was by modified Hodge test and the Carba-NP method. Isolates were investigated for the most common carbapenemases encoding genes bla(KPC), bla(NDM), and bla(OXA-48) using multiplex PCR. Molecular typing of carbapenem-resistant isolates was done by ERIC-PCR followed by sequencing of common resistance genes. The overall rate of CRBSI in our study was 3.6 per 1000 central venous catheter (CVC) days. Among 57 Gram-negative CRBSI isolates, Klebsiella pneumoniae (K. pneumoniae) was the most frequently isolated (27/57; 47.4%), of which more than 70% were resistant to Meropenem. Phenotypic tests for carbapenemases showed that 37.9% of isolates were positive by modified Hodge test and 63.8% by Carba-NP detection. Multiplex PCR assay detected the bla(NDM) in 28.6% of the isolates and bla(KPC) in 26.8%, bla(NDM) and bla(KPC) were detected together in the same isolate in 5.6%, while bla(OXA-48)-like were not detected. ERIC-PCR detected limited genetic relatedness between K. pneumoniae isolates. Elevated resistance rates were observed to all antibiotics including carbapenems among K. pneumoniae isolates causing CRBSI. ERIC-PCR showed that the resistant isolates were mainly polyclonal. Our results call for reinforcement of antimicrobial stewardship and measures to prevent CRBSI.201829936619
5193140.9903Antibiotic resistance genes prediction via whole genome sequence analysis of Stenotrophomonas maltophilia. BACKGROUND: Stenotrophomonas maltophilia (S. maltophilia) is the first dominant ubiquitous bacterial species identified from the genus Stenotrophomonas in 1943 from a human source. S. maltophilia clinical strains are resistance to several therapies, this study is designed to investigate the whole genome sequence and antimicrobial resistance genes prediction in Stenotrophomonas maltophilia (S. maltophilia) SARC-5 and SARC-6 strains, isolated from the nasopharyngeal samples of an immunocompromised patient. METHODS: These bacterial strains were obtained from Pakistan Institute of Medical Sciences (PIMS) Hospital, Pakistan. The bacterial genome was sequenced using a whole-genome shotgun via a commercial service that used an NGS (Next Generation Sequencing) technology called as Illumina Hiseq 2000 system for genomic sequencing. Moreover, detailed in-silico analyses were done to predict the presence of antibiotic resistance genes in S. maltophilia. RESULTS: Results showed that S. maltophilia is a rare gram negative, rod-shaped, non sporulating bacteria. The genome assembly results in 24 contigs (>500 bp) having a size of 4668,850 bp with 65.8% GC contents. Phylogenetic analysis showed that SARC-5 and SARC-6 were closely related to S. maltophilia B111, S. maltophilia BAB-5317, S. maltophilia AHL, S. maltophilia BAB-5307, S. maltophilia RD-AZPVI_04, S. maltophilia JFZ2, S. maltophilia RD_MAAMIB_06 and lastly with S. maltophilia sp ROi7. Moreover, the whole genome sequence analysis of both SARC-5 and SARC-6 revealed the presence of four resistance genes adeF, qacG, adeF, and smeR. CONCLUSION: Our study confirmed that S. maltophilia SARC-5 and SARC-6 are one of the leading causes of nosocomial infection which carry multiple antibiotic resistance genes.202438128408
1476150.9903Evaluation of the BioFire FilmArray Pneumonia Panel for rapid detection of respiratory bacterial pathogens and antibiotic resistance genes in sputum and endotracheal aspirate specimens. OBJECTIVES: The performance of the investigational-use-only version of the BioFire FilmArray Pneumonia Panel (FA-Pneumo), a high-order nested multiplex PCR, was evaluated for the detection of typical respiratory bacterial pathogens and antibiotic resistance genes in sputa and endotracheal aspirate (ETA) specimens. METHODS: Thirty-one sputa and 69 ETA specimens were analyzed. The diagnostic performance of FA-Pneumo was assessed using routine microbiological methods as the reference standard. RESULTS: Overall sensitivity and specificity for organism detection using FA-Pneumo were 98.5% and 76.5%, respectively. The sensitivity for each pathogen was 100%, except for Klebsiella aerogenes, and the range of specificity was 83.3-99.0%. FA-Pneumo detected antimicrobial resistance genes in 17 out of 18 specimens (94.4%) that were resistant by antimicrobial susceptibility testing. FA-Pneumo additionally detected 25 resistance genes in 22 specimens, and sequencing for the presence of resistance genes confirmed the majority of these results (20/25, 80%). Semi-quantitative analysis of bacterial nucleic acid amounts by FA-Pneumo revealed that 88.2% of the identified bacteria (67/76) with ≥10(6) copies/ml also gave culture-positive results with significant amounts of bacteria. CONCLUSIONS: FA-Pneumo is a rapid test with high sensitivity for the detection of bacteria and antimicrobial resistance genes in sputum and ETA specimens and could aid in determining antibiotic therapy.202032179139
2187160.9903Multicentre investigation of pathogenic bacteria and antibiotic resistance genes in Chinese patients with acute exacerbation of chronic obstructive pulmonary disease. OBJECTIVE: A prospective observational study to investigate the distribution and antimicrobial resistance of pathogenic bacteria in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) in Beijing, China. METHODS: Patients with AECOPD were recruited from 11 general hospitals. Sputum specimens were cultured and bacteria identified. Antibiotic susceptibility was determined for each isolate, and presence of antibiotic resistance genes was evaluated using polymerase chain reaction. RESULTS: Pathogenic bacteria were isolated from 109/318 patients (34.28%); 124 isolates of 22 pathogenic bacterial species were identified, including Klebsiella pneumoniae (16.94%), Pseudomonas aeruginosa (16.94%), Acinetobacter baumannii (11.29%), Streptococcus pneumoniae (8.87%), and Staphylococcus aureus (7.26%). S. aureus was sensitive to tigecycline, teicoplanin, vancomycin and linezolid but resistant to penicillin and levofloxacin. K.pneumoniae, P. aeruginosa, A. baumannii and E. coli were susceptible to amikacin and cefoperazone. CONCLUSIONS: K. pneumoniae and P. aeruginosa are the most common pathogenic bacteria in AECOPD cases in Beijing, China. Our antibiotic resistance findings may be helpful in selecting antibiotic therapy.201526152913
1244170.9902Identification of antibiotic resistance genes in Escherichia coli from subclinical mastitis milk in dairy cows and goats, East Java Province. Antibiotics are still used to treat mastitis in dairy cows in Indonesia. This study aimed to analyse antibiotic resistance genes in Escherichia coli (E. coli) from subclinical mastitis milk in East Java Province, Indonesia. The samples consisted of subclinical mastitis milk from cows and goats. A total of 592-quarter cow's milk and 71 goat's milk samples from both halves of the udder were collected from 67 farms in Lumajang, Banyuwangi, Malang, Sidoarjo, Jember, Pasuruan, Probolinggo, and Mojokerto. Subclinical mastitis samples were screened using the California mastitis test (CMT). E. coli was identified by phenotypic and genotypic methods. E. coli was confirmed with a primer specific to the polymerase chain reaction (PCR) technique. Gene resistance of E. coli was tested using the multiplex-PCR (mPCR) technique with primers encoding the genes temoneira enzyme (TEM), oxacillinase (OXA), sulfhydryl variable (SHV), and cefotaximase-munich IV (CTX-M IV). These genes were chosen because mastitis treatment generally uses oxacilline and β-lactam antibiotics. All data obtained were analysed descriptively. The results show that six isolates of E. coli (46.15%) carried a single resistance gene (TEM or SHV) and two isolates (33.33%) were confirmed as multiple drug-resistant organisms (MDROs) (TEM and SHV). The resistance genes were found in samples originating from Blitar, Banyuwangi, Lumajang, and Pasuruan Regencies. This research implies that antibiotic-resistance genes found in E. coli on certain farms are dangerous and may allow gene transmission to other bacteria that make treatment for mastitis or other bacterial infections ineffective.202438550619
1468180.9902Phenotypic and Molecular Characterization of Multidrug Resistant Klebsiella pneumoniae Isolated from Different Clinical Sources in Al-Najaf Province-Iraq. BACKGROUND AND OBJECTIVE: Burns infections and urinary tract infections are the most important prevalent diseases in Asian countries, such as Iraq. Klebsiella pneumoniae is one of the most important bacteria cause this type of infections especially in hospitals. Therefore, the aim of this study was to investigate the prevalence of multi-drug resistance K. pneumoniae and extended-spectrum beta-lactamases producing K. pneumoniae isolates from inpatients with urinary tract infection and burns infections in Al-Kufa hospital in Al-Najaf province, Iraq. MATERIALS AND METHODS: A total of 285 clinical samples were collected from in-patients infected with urinary tract infection (141 urine samples) and burns infections (144 burns swabs). Fourteen different antibiotics were used by disc diffusion method and 13 antimicrobials resistance genes were used by PCR technique. RESULTS: A total of 43 K. pneumoniae strains were isolated. The highest resistance rate was observed for amoxicillin 25 μg and amoxicillin+clavulanic acid 20+10 μg (97.67%) while the lowest resistance rate was observed for imipenem 10 μg (9.30%). The most common resistance associated-genes were blaSHV (86.04%) and at lower prevalence were IMP (9.30%). CONCLUSION: Klebsiella pneumoniae strains isolated from burns infections were more virulent than those isolated from urinary tract infections.201729023034
1402190.9902Detection of β-lactam resistance genes in Gram-negative bacteria from positive blood cultures using a microchip-based molecular assay. BACKGROUND: Accurate detection of β-lactam resistance genes in bloodstream infections is critical for guiding antimicrobial therapy. This study evaluates the Alifax Gram-negative resistance (GNR) microchip assay for detecting β-lactam resistance genes directly from positive blood cultures (PBCs) for Gram-negative (GN) bacteria, including Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter baumannii. METHODS: Simulated (n=146) and clinical (n=106) GN-PBC samples were tested for bla (KPC), bla (VIM), bla (NDM), bla (IMP), bla (OXA-23)-like, bla (OXA-48)-like, bla (SHV)-ESBL, bla (CTX-M-1/9) group, and bla (CMY-2)-like genes using the GNR microchip assay. Whole-genome sequencing (WGS) served as the reference assay for simulated samples and, selectively, for clinical samples. The bioMérieux BioFire Blood Culture Identification 2 (BCID2) panel assay was used as a comparator for clinical samples. RESULTS: The GNR microchip assay correctly identified 203 (99.5%) of 204 β-lactam resistance genes in simulated samples. One sample tested false negative for a bla (SHV)-ESBL gene but true positive for a bla (KPC) gene. In clinical samples, GNR results were concordant with BCID2 for 113 (100%) of 113 genes included in both assays. Additionally, the GNR assay detected bla (CMY-2) -like (n=6), bla (OXA-23)-like (n=5), and bla (SHV)-ESBL (n=2), which are not targeted by BCID2, all confirmed by WGS. In two β-lactam-resistant P. aeruginosa samples but negative by the GNR assay, WGS confirmed the absence of acquired β-lactam resistance genes, suggesting alternative resistance mechanisms. CONCLUSION: The GNR microchip assay demonstrated high concordance and broader β-lactam resistance gene coverage compared to BCID2, supporting its potential role in routine diagnostics. Further validation in larger, prospective studies is warranted.202540529307