scholarly journals 643. Comparison of Multiplex Polymerase Chain Reaction (PCR) and Routine Culture for the Detection of Respiratory Pathogens in Pneumonia Patients

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S297-S297
Author(s):  
Emad Abu Sitta ◽  
Nicole Hubbard ◽  
Geehan Suleyman
Keyword(s):  
Resistance Genes ◽  
Standard Procedure ◽  
Meca Gene ◽  
Bacterial Pathogen ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Use ◽  
Respiratory Pathogens ◽  
Culture Methods ◽  
Radiological Criteria ◽  
Two Samples

Abstract Background The identification of causative pathogens in pneumonia can be challenging, and conventional culture methods can take up to 72 hours. However, rapid microbiologic tests identify organisms within hours. The Biofire®Filmarray ((bioMérieux, North Carolina) Pneumonia Panel was recently approved by the FDA. The multiplex PCR system identifies 33 targets from sputum and bronchoalveolar (BAL) samples, which include 18 bacteria, 8 viruses, and 7 antibiotic resistance genes. The purpose is to compare the panel to routine culture methods for the detection of respiratory pathogens in patients with pneumonia in a 794-bed teaching hospital in northwest Ohio. Methods We retrospectively screened all hospitalized intensive care unit patients who met clinical and radiological criteria of pneumonia using electronic medical records between November 2018 and February 2019. Adult patients who had respiratory cultures collected within 7 days were included. Repeat specimens were excluded. Routine cultures were performed using the laboratory’s standard procedure, and Pneumonia Panel testing was performed according to manufacturer instructions. Results Fifty-nine respiratory or 13 BAL and 46 sputum specimens were evaluated. There was no discrepancy between culture and PCR in 63% (37/59) samples. One (8%) BAL and 10 (22%) sputum specimens had additional pathogens detected by PCR. There was a discrepancy between culture and PCR in four (31%) BAL and seven (15%) sputum samples. The largest discrepancy was noted amongst Serratia marcescens (4/59 or 7%) and Haemophilus influenzae (6/59 or 10%) species. Only one sputum culture had Legionella detected by PCR. Additionally, 17 specimens had a virus detected either alone or with another bacterial pathogen by PCR. For the resistance genes, KPC was detected by PCR but not by Modified Carbapenem Inactivation Method (mCIM) test. The mecA gene was detected in six of seven (86%) of methicillin-resistant Staphylococcus aureus (MRSA) isolates. CTX-M was detected in Serratia and Klebsiella pneunomiae in two samples; however, the organisms were not isolated in culture. Conclusion The Pneumonia Panel can identify additional bacteria that did not grow in culture. This panel can rapidly identify pathogens and potentially reduce unnecessary antibiotic use. Disclosures All authors: No reported disclosures

Quintuplex PCR to Detect Antibiotic Resistance Genes in Streptococcus pneumoniae

2016 ◽  
Vol 1 (2) ◽  
pp. 22 ◽  
Author(s):  
Navindra Kumari Palanisamy ◽  
Parasakthi Navaratnam ◽  
Shamala Devi Sekaran
Keyword(s):  
Antibiotic Resistance ◽  
Streptococcus Pneumoniae ◽  
Resistance Genes ◽  
Bacterial Pathogen ◽  
Antibiotic Resistance Genes ◽  
Pcr Amplification ◽  
Penicillin Resistance ◽  
Penicillin Binding Proteins ◽  
Efflux Mechanism ◽  
Mic Value

Introduction: Streptococcus pneumoniae is an important bacterial pathogen, causing respiratory infection. Penicillin resistance in S. pneumoniae is associated with alterations in the penicillin binding proteins, while resistance to macrolides is conferred either by the modification of the ribosomal target site or efflux mechanism. This study aimed to characterize S. pneumoniae and its antibiotic resistance genes using 2 sets of multiplex PCRs. Methods: A quintuplex and triplex PCR was used to characterize the pbp1A, ermB, gyrA, ply, and the mefE genes. Fifty-eight penicillin sensitive strains (PSSP), 36 penicillin intermediate strains (PISP) and 26 penicillin resistance strains (PRSP) were used. Results: Alteration in pbp1A was only observed in PISP and PRSP strains, while PCR amplification of the ermB or mefE was observed only in strains with reduced susceptibility to erythromycin. The assay was found to be sensitive as simulated blood cultures showed the lowest level of detection to be 10cfu. Conclusions: As predicted, the assay was able to differentiate penicillin susceptible from the non-susceptible strains based on the detection of the pbp1A gene, which correlated with the MIC value of the strains.

Retail Ready-to-Eat Food as a Potential Vehicle for Staphylococcus spp. Harboring Antibiotic Resistance Genes

2014 ◽  
Vol 77 (6) ◽  
pp. 993-998 ◽  
Author(s):  
WIOLETA CHAJĘCKA-WIERZCHOWSKA ◽  
ANNA ZADERNOWSKA ◽  
BEATA NALEPA ◽  
MAGDA SIERPI´NSKA ◽  
ŁUCJA ŁANIEWSKA-TROKENHEIM
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Meca Gene ◽  
Antibiotic Resistance Genes ◽  
Tetracycline Resistance ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Antibiotic Resistant ◽  
Cured Meats ◽  
Staphylococcus Spp

Ready-to-eat (RTE) food, which does not need thermal processing before consumption, could be a vehicle for the spread of antibiotic-resistant microorganisms. As part of general microbiological safety checks, staphylococci are routinely enumerated in these kinds of foods. However, the presence of antibiotic-resistant staphylococci in RTE food is not routinely investigated, and data are only available from a small number of studies. The present study evaluated the pheno- and genotypical antimicrobial resistance profile of Staphylococcus spp. isolated from 858 RTE foods (cheeses, cured meats, sausages, smoked fishes, salads). Of 113 strains isolated, S. aureus was the most prevalent species, followed by S. xylosus, S. saprophyticus, and S. epidermidis. More than half (54.9%) of the isolates were resistant to at least one class of tested antibiotic; of these, 35.4% of the strains were classified as multidrug resistant. Most of the isolates were resistant to cefoxitin (49.6%), followed by clindamycin (39.3%), tigecycline (27.4%), quinupristin-dalfopristin (22.2%), rifampin (20.5%), tetracycline (17.9%), and erythromycin (8.5%). All methicillin-resistant staphylococci harbored the mecA gene. Among the isolates resistant to at least one antibiotic, 38 harbored tetracycline resistance determinant tet(M), 24 harbored tet(L), and 9 harbored tet(K). Of the isolates positive for tet(M) genes, 34.2% were positive for the Tn916-Tn1545–like integrase family gene. Our results indicated that retail RTE food could be considered an important route for the transmission of antibiotic-resistant bacteria harboring multiple antibiotic resistance genes.

scholarly journals Comparison of Antibiotic Resistance Mechanisms in Antibiotic-Producing and Pathogenic Bacteria

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3430 ◽  
Author(s):  
Hiroshi Ogawara
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Pathogenic Bacteria ◽  
Early Stage ◽  
Antibiotic Resistance Genes ◽  
Gene Clusters ◽  
Antibiotic Use ◽  
Resistance Mechanisms ◽  
Biosynthetic Gene Clusters ◽  
Points Of View

Antibiotic resistance poses a tremendous threat to human health. To overcome this problem, it is essential to know the mechanism of antibiotic resistance in antibiotic-producing and pathogenic bacteria. This paper deals with this problem from four points of view. First, the antibiotic resistance genes in producers are discussed related to their biosynthesis. Most resistance genes are present within the biosynthetic gene clusters, but some genes such as paromomycin acetyltransferases are located far outside the gene cluster. Second, when the antibiotic resistance genes in pathogens are compared with those in the producers, resistance mechanisms have dependency on antibiotic classes, and, in addition, new types of resistance mechanisms such as Eis aminoglycoside acetyltransferase and self-sacrifice proteins in enediyne antibiotics emerge in pathogens. Third, the relationships of the resistance genes between producers and pathogens are reevaluated at their amino acid sequence as well as nucleotide sequence levels. Pathogenic bacteria possess other resistance mechanisms than those in antibiotic producers. In addition, resistance mechanisms are little different between early stage of antibiotic use and the present time, e.g., β-lactam resistance in Staphylococcus aureus. Lastly, guanine + cytosine (GC) barrier in gene transfer to pathogenic bacteria is considered. Now, the resistance genes constitute resistome composed of complicated mixture from divergent environments.

scholarly journals The occurrence of antibiotic resistance genes in the microbiota of yak, beef and dairy cattle characterized by a metagenomic approach

2021 ◽  
Author(s):  
Weiwei Wang ◽  
Xiaojuan Wei ◽  
Lingyu Wu ◽  
Xiaofei Shang ◽  
Fusheng Cheng ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Antibiotic Resistance ◽  
Dairy Cattle ◽  
Resistance Genes ◽  
Animal Feed ◽  
Growth Promotion ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Use ◽  
Agricultural Animal ◽  
Metagenomic Approach

AbstractDrug resistance has been partly driven by the overuse of antimicrobials in agricultural animal feed. Better understanding of antibiotic resistance in bovine gut is needed to assess its potential effects based on metagenomic approach and analysis. In this study, we collected 40 fecal samples to explore drug resistance derived from antibiotic use in the bacterial community by an analysis of the diversities and differences of antibiotic-resistant genes (ARGs) in the gut microbiota from yak, beef, and dairy cattle. Overall, 1688 genes were annotated, including 734 ARG subtypes. The ARGs were related to tetracyclines, quinolones, β-lactam, and aminoglycosides, in accordance with the antibiotics widely used in the clinic for humans or animals. The emergence, prevalence, and differences in resistance genes in the intestines of yaks, beef, and dairy cattle may be caused by the selective pressure of different feeding patterns, where yaks were raised without antibiotics for growth promotion. In addition, the abundance of ARGs in yak was lower than in beef and dairy cattle, whereas the abundance of integron, a kind of mobile genetic elements (MGEs) was higher in yaks than those in beef and dairy cattle. Furthermore, the results of this study could provide the basis for a comprehensive profile of various ARGs among yak, beef, and dairy cattle in future.

scholarly journals The perfect condition for the rising of superbugs: person-to-person contagion and antibiotic use are the key factors responsible for the positive correlation between antibiotic resistance gene diversity and virulence gene diversity in human metagenomes

2020 ◽  
Author(s):  
Célia P. F. Domingues ◽  
João S. Rebelo ◽  
Teresa Nogueira ◽  
Joël Pothier ◽  
Francisca Monteiro ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Loss Rate ◽  
Gene Diversity ◽  
Recent Observation ◽  
Antibiotic Resistance Gene ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Use ◽  
Resistant Bacteria ◽  
Positive Correlation

1.AbstractThis study aims to understand the cause of the recent observation that humans with a higher diversity of virulence genes in their metagenomes tend to be precisely those with higher diversity of antibiotic-resistance genes. We simulated the transferring of virulence and antibiotic-resistance genes in a community of interacting people where some take antibiotics. The diversities of the two genes types became positively correlated whenever the contagion probability between two people was higher than the probability of losing resistant genes. However, no such positive correlations arise if no one takes antibiotics. This finding holds even under changes of several simulations’ parameters, such as the relative or total diversity of virulence and resistance genes, the contagion probability between individuals, the loss rate of resistance genes, or the social network type. Because the loss rate of resistance genes may be shallow, we conclude that the contagion between people and antibiotic usage is the leading cause of establishing the positive correlation mentioned above. Therefore, antibiotic use and something as prosaic as the contagion between people may facilitate the emergence of virulent and multi-resistant bacteria in people’s metagenomes with a high diversity of both gene types. These superbugs may then circulate in the community.

scholarly journals Microbiota analysis of rural and urban surface waters and sediments in Bangladesh identifies human waste as driver of antibiotic resistance

2021 ◽  
Author(s):  
Ross Stuart McInnes ◽  
Md. Hassan uz-Zaman ◽  
Imam Taskin Alam ◽  
Siu Fung Stanley Ho ◽  
Robert A. Moran ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Surface Waters ◽  
Water Samples ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Use ◽  
Urban Surface ◽  
Resistant Bacteria ◽  
Antibiotic Resistant Bacteria ◽  
Antibiotic Resistant

AbstractIn many low- and middle-income countries antibiotic resistant bacteria spread in the environment due to inadequate treatment of wastewater and the poorly regulated use of antibiotics in agri- and aquaculture. Here we characterised the abundance and diversity of antibiotic-resistant bacteria and antibiotic resistance genes in surface waters and sediments in Bangladesh through quantitative culture of Extended-Spectrum Beta-Lactamase (ESBL)-producing coliforms and shotgun metagenomics. Samples were collected from highly urbanised settings (n = 7), from rural ponds with a history of aquaculture-related antibiotic use (n = 11) and from rural ponds with no history of antibiotic use (n = 6). ESBL-producing coliforms were found to be more prevalent in urban samples than in rural samples. Shotgun sequencing showed that sediment samples were dominated by the phylum Proteobacteria (on average 73.8% of assigned reads), while in the water samples Cyanobacteria (on average 60.9% of assigned reads) were the predominant phylum. Antibiotic resistance genes were detected in all samples, but their abundance varied 1,525-fold between sites, with the highest levels of antibiotic resistance genes being present in urban surface water samples. We identified an IncQ1 sulphonamide resistance plasmid ancestral to the widely studied RSF1010 in one of the urban water samples. The abundance of antibiotic resistance genes was significantly correlated (R2 = 0.73; P = 8.9 × 10−15) with the abundance of bacteria originating from the human gut, which suggests that the release of untreated sewage is a driver for the spread of environmental antibiotic resistance genes in Bangladesh, particularly in highly urbanised settings.ImportanceLow- and middle-income countries (LMICs) have higher burdens of multidrug-resistant infections than high-income countries and there is thus an urgent need to elucidate the drivers of the spread of antibiotic-resistant bacteria in LMICs. Here we study the diversity and abundance of antibiotic resistance genes in surface water and sediments from rural and urban settings in Bangladesh. We found that urban surface waters are particularly rich in antibiotic resistance genes, with a higher number of them associated with plasmids indicating that they are more likely to spread horizontally. The abundance of antibiotic resistance genes was strongly correlated with the abundance of bacteria that originate from the human gut, suggesting that uncontrolled release of human waste is a major driver for the spread of antibiotic resistance in the urban environment. Improvements in sanitation in LMICs may thus be a key intervention to reduce the dissemination of antibiotic resistant bacteria.

scholarly journals Biocide use, integrons and novel genetic elements

2010 ◽  
Vol 31 (4) ◽  
pp. 192 ◽  
Author(s):  
Michael R Gillings
Keyword(s):  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Use ◽  
Consumer Products ◽  
Mobile Dna ◽  
Bacterial Strains ◽  
Class 1 Integron ◽  
Genetic Elements ◽  
Inappropriate Use ◽  
Selection Of

Resistance to antibiotics threatens our ability to control bacterial pathogens. It is clear that the persistence of cells containing resistance determinants is promoted by the strong selective pressure imposed by antibiotic use. This problem has been exacerbated by inappropriate and excessive use of antibiotics in both medicine and animal production. Concern has also been raised that inappropriate use of biocides contributes to the selection of resistant bacterial strains. This may occur because detoxification mechanisms for biocides and antibiotics are shared, or via selection for biocide resistance genes that are physically linked to antibiotic resistance genes and their mobile DNA vectors. In this brief review I will illustrate the latter phenomenon using the evolutionary history of the class 1 integron as an example, and then examine whether the increasing trend towards indiscriminate use of biocides in homes and consumer products might result in the selection of novel genetic elements that will have negative and unpredictable consequences for human health.

scholarly journals Plant Agricultural Streptomycin Formulations Do Not Carry Antibiotic Resistance Genes

2009 ◽  
Vol 53 (7) ◽  
pp. 3173-3177 ◽  
Author(s):  
Fabio Rezzonico ◽  
Virginia O. Stockwell ◽  
Brion Duffy
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Extraction Procedure ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Use ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Bacterial Disease ◽  
Pome Fruit ◽  
Fruit Orchards

ABSTRACT Streptomycin is used in plant agriculture for bacterial disease control, particularly against fire blight in pome fruit orchards. Concerns that this may increase environmental antibiotic resistance have led to bans or restrictions on use. Experience with antibiotic use in animal feeds raises the possible influence of formulation-delivered resistance genes. We demonstrate that agricultural streptomycin formulations do not carry producer organism resistance genes. By using an optimized extraction procedure, Streptomyces 16S rRNA genes and the streptomycin resistance gene strA were not detected in agricultural streptomycin formulations. This diminishes the likelihood for one potential factor in resistance development due to streptomycin use.

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