Development and Application of a Novel Nucleic Amplification Kit on Rapid Detection of Various Types of Staphylococci Strains

2014 ◽  
Vol 618 ◽  
pp. 288-292
Author(s):  
Yang Deng ◽  
Yan Rui Liang ◽  
Lin Li ◽  
Bing Li ◽  
Jian Yu Su
Keyword(s):  
Antibiotic Resistance ◽  
Predictive Value ◽  
Rapid Detection ◽  
Detection Rate ◽  
Lamp Method ◽  
Gram Negative ◽  
Optimal Reaction Condition ◽  
Optimal Reaction ◽  
Reference Strains ◽  
Antibiotic Resistance Determinant

A loop-mediated isothermal amplification (LAMP) method for rapid detection of various staphylococci strains and associated antibiotic resistance determinant had been developed and evaluated in this study. Six primers, including outer primers, inner primers and loop primers, were specially designed for recognizing eight distinct sequences on four targets: 16Sr RNA,femA,mecA, andorfX. Twenty-seven reference strains, including various species of gram-negative and-positive isolates, were included in this study to evaluate and optimize LAMP assays. The optimal reaction condition was found to be 65°C for 45 min, with detection limits at 100 fg DNA/tube and 10 CFU/reaction for 16S rRNA, 100 fg DNA/tube and 10 CFU/reaction forfemA, 1 pg DNA/tube and 100 CFU/reaction formecA, 10 DNA/tube and 10 CFU/reaction fororfX, respectively. Application of LAMP assays were performed on 166 various types of staphylococci isolates, the detection rate of LAMP assays for the 16Sr RNA,femA,mecA, andorfXwas 100% (166/166), 98.5% (64/65), 94.3% (66/70), and 98.6% (69/70) and the negative predictive value (NPV) was 100%, 98.1%, 92.3%, and 92.7% respectively; with a 100% positive predictive value (PPV) for all three targets. In conclusion, LAMP assays were demonstrated to be useful and powerful tools for rapid detection of various staphylococci strains.

scholarly journals 2175. Rapid Detection of Carbapenemase Producing Organisms Directly from Blood Cultures Positive for Gram-Negative Bacilli

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S738-S738
Author(s):  
William Stokes ◽  
Johann Pitout ◽  
Lorraine Campbell ◽  
Deirdre Church ◽  
Dan Gregson
Keyword(s):  
Predictive Value ◽  
Rapid Detection ◽  
Blood Cultures ◽  
Gram Negative ◽  
Carbapenem Resistant ◽  
Direct Testing ◽  
Appropriate Treatment ◽  
Risk Patients ◽  
Health Zone ◽  
Sensitivity Specificity

Abstract Background The rapid detection of carbapenemase-producing organisms (CPOs) directly from blood cultures (BC) positive for Gram-negative bacilli (GNB) may accelerate the appropriate treatment of at-risk patients. Our objective was to evaluate the performance of two commercial assays in the rapid detection of CPOs directly from BC positive for GNB. Methods BC positive for GNB, taken from patients within the Calgary Health Zone over a 3 month period, were tested for the presence of CPOs with βCARBA® and NG-Test® CARBA 5. A subset of sterile BC samples was seeded with multi-drug-resistant (MDR) GNB. BC were incubated using the Bact-Alert™ system. Positive BC from clinical and seeded samples was tested directly with βCARBA and CARBA 5 from BC pellets processed for direct testing using an ammonium chloride lysis and wash method. Sensitivity, specificity, negative predictive value (NPV) and positive predictive value (PPV) were calculated with 95% confidence intervals for binomial proportions. Results 65 samples were tested (30 clinical, 35 seeded). Seeded samples included 1 GES, 4 IMP, 6 KPC, 1 co-producing KPC and NDM, 9 OXA, 4 VIM, 5 NDM, and 5 non-CPO carbapenem-resistant organisms. βCARBA had a sensitivity, specificity, NPV and PPV of 100% (88.4% - 100%), 65.7% (47.8–80.9%), 100%, and 71.4% (61.3%–79.8%), respectively. CARBA 5 had a sensitivity, specificity, NPV and PPV of 90.0% (73.5%–97.9%), 100% (90.0%–100%), 92.1% (80.0%–97.2%), and 100%. When excluding GES, which is known not to be detected by CARBA 5, sensitivity and NPV increased to 93.1% (77.2%–99.2%) and 93.1% (78.0%–98.1%), respectively. False negatives for βCARBA occurred with 1 VIM-1 and IMP-14. Conclusion This study demonstrates that the detection of CPOs directly from positive BC can be accurately achieved. βCARBA had excellent sensitivity but suffered from poor specificity. CARBA 5 had good sensitivity and specificity but is unable to detect certain CPOs. Testing positive BC directly using βCARBA and/or CARBA 5 may be useful in rapidly detecting CPOs. Results of direct testing from the CARBA5 assay would quickly identify patients amenable to treatment with avibactam combination compounds. Disclosures All authors: No reported disclosures.

Rapid Detection of Antibiotic Resistance in Gram-Negative Bacteria Through Assessment of Changes in Cellular Morphology

2017 ◽  
Vol 23 (2) ◽  
pp. 157-162 ◽  
Author(s):  
Fátima Otero ◽  
Rebeca Santiso ◽  
Maria Tamayo ◽  
José Luis Fernández ◽  
Germán Bou ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Rapid Detection ◽  
Cellular Morphology ◽  
Gram Negative Bacteria ◽  
Gram Negative

Extended Spectrum Beta Lactamase (ESBL), bla TEM,bla SHVand bla CTX-M,resistance genes in Community and Healthcare Associated Gram Negative Bacteria from Osun State, Nigeria

2020 ◽  
Vol 20 ◽  
Author(s):  
Ganiyat Shitta ◽  
Olufunmilola Makanjuola ◽  
Olusolabomi Adefioye ◽  
Olugbenga Adekunle Olowe
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Gram Negative Bacteria ◽  
Beta Lactamase ◽  
Multiple Antibiotic Resistance ◽  
Esbl Production ◽  
Gram Negative ◽  
Extended Spectrum Beta Lactamase ◽  
Healthcare Associated ◽  
Esbl Producers

Background: Extended Spectrum Beta Lactamase (ESBL) production in gram negative bacteria confers multiple antibiotic resistance, adversely affecting antimicrobial therapy in infected individuals. ESBLs result from mutations in β-lactamases encoded mainly by the bla TEM,bla SHVand bla CTX-Mgenes. The prevalence of ESBL producing bacteria has been on the increase globally especially its upsurge among isolates from community-acquired infections. Aim: To determine ESBL prevalence and identify ESBL genes among clinical isolates in Osun State, Nigeria. Material and Methods: A cross-sectional study was carried out from August 2016 –July 2017 in Osun State, Nigeria. Three hundred and sixty Gram negative bacteria recovered from clinical samples obtained from both community and healthcare associated infections were tested. They included147 Escherichia coli(40.8%), 116 Klebsiella spp(32.2%), 44 Pseudomo-nas aeruginosa(12.2%) and23 Proteus vulgaris (6.4%) isolates. Others were Acinetobacter baumannii, Serratia rubidae, Citrobacter spp, Enterobacter spp and Salmonella typhi. Disk diffusion antibiotic susceptibility testing was carried out, isolates were screened for ESBL production and confirmed using standard laboratory procedures. ESBLs resistance genes were identified by Polymerase Chain Reaction (PCR). Results: All isolates demonstrated multiple antibiotic resistance. Resistance to ampicillin, amoxicillin with clavulanate and erythromycin was 100%, whereas resistance to Imipenem was very low (5.0%). : Overall prevalence of ESBL producers was 41.4% with Klebsiellaspp as the highest ESBL producing Enterobacteriacaea. ESBL producers were more prevalent among the hospital pathogens than community pathogens, 58% vs 29.5% (p=0.003). ESBL genes were detected in all ESBL producers with the blaCTX-Mgene predominating (47.0%) followed by blaTEM(30.9%) and blaSHVgene was the least, 22.1%. The blaCTX-Mgene was also the most prevalent in the healthcare pathogens (62%) but it accounted for only 25% in those of community origin. Conclusion: A high prevalence of ESBL producing gram negative organisms occurs both in healthcare and in the community in our environment with the CTX-M variant predominating. Efforts to control spread of these pathogens should be addressed.

scholarly journals A113 ANNUAL COLONOSCOPY VOLUME IS NOT PREDICTIVE OF COLONOSCOPY QUALITY - FINDINGS FROM THE SOUTHWEST ONTARIO COLONOSCOPY COHORT

2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 92-93
Author(s):  
M Sey ◽  
O Siddiqi ◽  
C McDonald ◽  
S cocco ◽  
Z Hindi ◽  
...  
Keyword(s):  
Predictive Value ◽  
Detection Rate ◽  
Primary Outcome ◽  
Case Mix ◽  
Operating Characteristics ◽  
Polyp Detection ◽  
Cecal Intubation ◽  
Polyp Detection Rate ◽  
Secondary Outcomes ◽  
Colonoscopy Quality

Abstract Background Performing a minimum number of colonoscopies annually has been proposed by some jurisdictions as a requirement for maintaining privileges. However, this practice is supported by limited evidence. Aims The objective of this study was to determine if annual colonoscopy volume was associated with colonoscopy quality metrics. Methods A population-based study was performed using the Southwest Ontario Colonoscopy cohort, which consists of all adult patients who underwent colonoscopy between April 2017 and Oct 2018 at 21 academic and community hospitals within the health region. Data were collected through a mandatory quality assurance form completed after each procedure and pathology reports were manually reviewed. Physician annualized colonoscopy volumes were compared by correlation analysis to each quality-related outcome, by means of the area under the receiver operating characteristics curve (AUROC), and logistic regression. The prognostic value of colonoscopy volume was also adjusted for case-mix and potential confounders in separate regression analyses for each outcome. The primary outcome was ADR. Secondary outcomes were polyp detection rate (PDR), sessile serrated polyp detection rate (SSPDR), and cecal intubation. Results A total of 47,195 colonoscopies were performed by 75 physicians (37.5% by gastroenterologists, 60% by general surgeons, 2.5% others). There were no clear relationships between annual colonoscopy volumes and study outcomes. Colonoscopy volume was not associated with ADR (OR 1.03, 95% CI 0.96–1.10, p=0.48) and corresponded to an AUROC not significantly different from the null (AUROC 0.52, 95% CI 0.43–0.61, p=0.65). Multi-variable regression adjusting for case-mix also demonstrated no predictive value of annual colonoscopy volume for the primary outcome (OR 1.03, 95% CI 0.94–1.12, p=0.55). Similarly, analyses of secondary outcomes failed to find an association between colonoscopy volume and PDR, SSPDR, or cecal intubation (Table 1). Conclusions Annual colonoscopy volumes do not predict ADR, PDR, SSPDR, or cecal intubation rate. Results of unconditional and conditional approaches for examining the predictive value of annual colonoscopy volume for quality related outcomes. Funding Agencies None

scholarly journals Correlation between the Antibiotic Resistance Genes and Susceptibility to Antibiotics among the Carbapenem-Resistant Gram-Negative Pathogens

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 255
Author(s):  
Salma M. Abdelaziz ◽  
Khaled M. Aboshanab ◽  
Ibrahim S. Yahia ◽  
Mahmoud A. Yassien ◽  
Nadia A. Hassouna
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Efflux Pump ◽  
Antibiotic Resistance Genes ◽  
Multiple Drug ◽  
P Value ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Gram Negative ◽  
Carbapenem Resistant

In this study, the correlation between the antibiotic resistance genes and antibiotic susceptibility among the carbapenem-resistant Gram-negative pathogens (CRGNPs) recovered from patients diagnosed with acute pneumonia in Egypt was found. A total of 194 isolates including Klebsiella pneumoniae (89; 46%), Escherichia coli (47; 24%) and Pseudomonas aeruginosa (58; 30%) were recovered. Of these, 34 (18%) isolates were multiple drug resistant (MDR) and carbapenem resistant. For the K. pneumoniae MDR isolates (n = 22), blaNDM (14; 64%) was the most prevalent carbapenemase, followed by blaOXA-48 (11; 50%) and blaVIM (4; 18%). A significant association (p value < 0.05) was observed between the multidrug efflux pump (AcrA) and resistance to β-lactams and the aminoglycoside acetyl transferase gene (aac-6’-Ib) gene and resistance to ciprofloxacin, azithromycin and β-lactams (except for aztreonam). For P. aeruginosa, a significant association was noticed between the presence of the blaSHV gene and the multidrug efflux pump (MexA) and resistance to fluoroquinolones, amikacin, tobramycin, co-trimoxazole and β-lactams and between the aac-6’-Ib gene and resistance to aminoglycosides. All P. aeruginosa isolates (100%) harbored the MexAB-OprM multidrug efflux pump while 86% of the K. pneumoniae isolates harbored the AcrAB-TolC pump. Our results are of great medical importance for the guidance of healthcare practitioners for effective antibiotic prescription.

Direct Synthesis of 3,5-Dinitropyrazole Catalyzed by Newly Sulfated Nanosolid Superacid TiO2/ SO

2011 ◽  
Vol 391-392 ◽  
pp. 1296-1301
Author(s):  
Li Min Xi ◽  
Xin Xin Zhang
Keyword(s):  
Direct Synthesis ◽  
Sol Gel ◽  
Acid Base ◽  
Direct Transformation ◽  
Acid Base Titration ◽  
Optimal Reaction Condition ◽  
Superacid Catalyst ◽  
Average Size ◽  
Optimal Reaction ◽  
Xrd And Tem

The newly sulfated nanosolid superacid TiO2/SO4 prepared by sol-gel method was broadly characterized by acid base titration, XRD and TEM, which identified that the superfine solid TiO2/SO4 showing good dispersibility with average size of 27 nm belongs to kind of crystalline nanoparticles. With the help of the catalyst TiO2/SO4, the optimal reaction condition for direct transformation of pyrazole and nitrosonitric acid into 3, 5-Dinitropyrazole was n (pyrazol)=0.10 mol, m (TiO2/SO4 )＝1.5g, V(n-octannol)＝90mL, and V(nitrosonitric acid)＝50mL. Moreover, the optimal yield of the catalytic reaction reached up to 59.4% when the reaction time is 7 hours. The nanosolid superacid catalyst is still of high activity after regenerating eight times by calcination at 600。C.

scholarly journals Evaluation of the Carba NP Test for Rapid Detection of Carbapenemase-Producing Enterobacteriaceae and Pseudomonas aeruginosa

2013 ◽  
Vol 57 (9) ◽  
pp. 4578-4580 ◽  
Author(s):  
Nathalie Tijet ◽  
David Boyd ◽  
Samir N. Patel ◽  
Michael R. Mulvey ◽  
Roberto G. Melano
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Positive Predictive Value ◽  
Negative Predictive Value ◽  
Predictive Value ◽  
Rapid Detection ◽  
False Negative ◽  
Bacterial Extract ◽  
Content Type ◽  
Negative Results ◽  
False Negative Results

ABSTRACTThe Carba NP test was evaluated against a panel of 244 carbapenemase- and non-carbapenemase-producingEnterobacteriaceaeandPseudomonas aeruginosaisolates. We confirmed the 100% specificity and positive predictive value of the test, but the sensitivity and negative predictive value were 72.5% and 69.2%, respectively, and increased to 80% and 77.3%, respectively, using a more concentrated bacterial extract. False-negative results were associated with mucoid strains or linked to enzymes with low carbapenemase activity, particularly OXA-48-like, which has emerged globally in enterobacteria.

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