scholarly journals Development of multiplex polymerase chain reaction assay for rapid detection ofStaphylococcus aureusand selected antibiotic resistance genes in bovine mastitic milk samples

2011 ◽  
Vol 23 (5) ◽  
pp. 894-901 ◽  
Author(s):  
Jian Gao ◽  
Miro Ferreri ◽  
Xiu Q. Liu ◽  
Li B. Chen ◽  
Jing L. Su ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Rapid Detection ◽  
Multiplex Polymerase Chain Reaction ◽  
Antibiotic Resistance Genes ◽  
Chain Reaction ◽  
Milk Samples ◽  
Polymerase Chain ◽  
Mastitic Milk

scholarly journals Detection of selected antibiotic resistance genes using multiplex PCR assay in mastitis pathogens in the Czech Republic

2017 ◽  
Vol 86 (2) ◽  
pp. 167-174 ◽  
Author(s):  
Vladimir Pyatov ◽  
Irena Vrtková ◽  
Aleš Knoll
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Czech Republic ◽  
Resistance Genes ◽  
Multiplex Polymerase Chain Reaction ◽  
Antibiotic Resistance Genes ◽  
Chain Reaction ◽  
The Czech Republic ◽  
Polymerase Chain ◽  
Mastitis Pathogens

The aim of this research was to develop multiplex polymerase chain reaction assays for the detection of aminoglycoside (strA, strB), sulphonamide (sulI, sulII), tetracycline (tetA, tetB, tetK, tetM, tetO), macrolide and lincosamide (msrA, ermA, ermB, ermC, mefA/E) genes of resistance in mastitis pathogens (Escherichia coli, Staphylococcus aureus, Streptococcus uberis, Streptococcus agalactiae and Streptococcus dysgalactiae). Applying the established assays, we investigated the distribution of antibiotic resistance genes in the above mentioned species isolated from milk samples in the Czech Republic. Each assay consisted of seven pairs of primers. Six of them amplified fragments of antibiotic resistance genes and one pair a fragment of a species specific gene. Polymerase chain reaction conditions were optimized to amplify seven gene fragments simultaneously in one reaction. In total, 249 isolates were used, among which 111 were positive for E. coli, 52 for S. aureus and 86 for Streptococcus spp. The majority (60.2%) of bacteria carried at least one antibiotic resistance gene and 44.6% were multidrug-resistant. The designed multiplex polymerase chain reaction assays may be applied as diagnostic method to replace or complement standard techniques of antibiotic susceptibility testing in the mentioned pathogens.

scholarly journals Detection and Profiling of Antibiotic Resistance among Culturable Bacterial Isolates in Vended Food and Soil Samples

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Susan W. Muriuki ◽  
Johnstone O. Neondo ◽  
Nancy L. M. Budambula
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Risk Group ◽  
Antibiotic Resistance Genes ◽  
Soil Samples ◽  
Bacterial Isolates ◽  
Chain Reaction ◽  
Antibiotic Resistant ◽  
Polymerase Chain

The emergence and persistence of antibiotic resistance remain formidable health challenges. This study aimed at detecting and profiling antibiotic resistance of bacterial contaminants in vended food and the environment. Seventy antibiotic-resistant bacterial isolates were isolated from fried fish, African sausages, roasted meat, smokies, samosa, chips (potato fries), vegetable salads, and soil samples collected from Embu Town and Kangaru Market in Embu County, Kenya. The antibiotic susceptibility test, morphological and biochemical characterization, antibiosis assay, polymerase chain reaction-based detection of antibiotic resistance genes, and sequencing of the 16S rRNA gene were done. Analysis of variance on all measured data was done, and Tukey’s honest test was used to compare and separate mean diameters of zones inhibition. Resistance of bacterial isolates to antibiotics was chloramphenicol (90%), cefotaxime (84.29%), nalidixic acid (81.43%), tetracycline (77.14%), amoxicillin (72.86%), gentamycin (48.57%), streptomycin (32.86%), and trimethoprim + sulphamethoxazole (30%). Isolate KMP337, Salmonella spp., exhibited highly significant antibiosis against S. aureus recording a mean inhibition diameter and standard error (SE) of 16.33 ± 0.88 mm, respectively, at P=0.001. The 70 bacterial isolates belonged to Bacillus, Paraclostridium, Lysinibacillus, Virgibacillus, and Serratia genera. The study isolated Bacillus wiedmannii (KC75) which is a risk group 2 as well as Serratia marcescens (KMP95) and Bacillus anthracis (KS606) which are risk group 3 organisms. The presence of antibiotic resistance genes Tet A, BlaTEM, StrB, Dfr A, Amp, and FloR genes was confirmed by a polymerase chain reaction. Samples from Kangaru Market recorded a higher (88.57%) proportion of resistant isolates as compared to isolates from Embu Town (11.43%). The study confirmed the presence of antibiotic-resistant bacteria in vended fast food and the soil in Embu Town and Kangaru Market. This study calls for continuous monitoring of bacterial status and hygienic handling of vended food.

scholarly journals Transferring of Lactobacillus antibiotic resistant genes to Salmonella

2021 ◽  
pp. 145-151
Author(s):  
Muhammad Ashraf ◽  
Sajjad-ur- Rahman ◽  
Muhammad Jawad Bashir ◽  
Rizwan Aslam ◽  
Sultan Ali ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Real Time ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Growth Promoters ◽  
Chain Reaction ◽  
Antibiotic Resistant ◽  
Polymerase Chain

Antibiotic resistance is a worldwide issue and becoming more problematic due to extensive misuse of antibiotics. The present study was aimed to analyze role of Lactobacillus in transmission of antibiotic resistance genes (tetM, ermB, sul2) to Salmonella and verification of these genes by real time polymerase chain reaction. A total of thirty fecal samples (15 were indigenous and 15 were broilers) were collected and analyzed by real time polymerase chain reaction. The results indicated that there was high expression of antibiotic resistance genes in Lactobacillus in case of broiler chicken than indigenous ones indicating Lactobacillus as a reservoir of antibiotic resistance genes but found to be non-significant in transferring these genes to Salmonella. In conclusion, the excessive use of animal growth promoters in poultry assists in acquisition of antibiotic resistance genes by normal micro-biota. Keywords: Broiler, Non-significant, Antibiotic resistance, Real time polymerase chain

scholarly journals Competitiveness of Quantitative Polymerase Chain Reaction (qPCR) and Droplet Digital Polymerase Chain Reaction (ddPCR) Technologies, with a Particular Focus on Detection of Antibiotic Resistance Genes (ARGs)

2021 ◽  
Vol 1 (3) ◽  
pp. 426-444
Author(s):  
Sol Park ◽  
Anita Rana ◽  
Way Sung ◽  
Mariya Munir
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Antibiotic Resistance Genes ◽  
Chain Reaction ◽  
Viral Infectious Disease ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

With fast-growing polymerase chain reaction (PCR) technologies and various application methods, the technique has benefited science and medical fields. While having strengths and limitations on each technology, there are not many studies comparing the efficiency and specificity of PCR technologies. The objective of this review is to summarize a large amount of scattered information on PCR technologies focused on the two majorly used technologies: qPCR (quantitative polymerase chain reaction) and ddPCR (droplet-digital polymerase chain reaction). Here we analyze and compare the two methods for (1) efficiency, (2) range of detection and limitations under different disciplines and gene targets, (3) optimization, and (4) status on antibiotic resistance genes (ARGs) analysis. It has been identified that the range of detection and quantification limit varies depending on the PCR method and the type of sample. Careful optimization of target gene analysis is essential for building robust analysis for both qPCR and ddPCR. In our era where mutation of genes may lead to a pandemic of viral infectious disease or antibiotic resistance-induced health threats, this study hopes to set guidelines for meticulous detection, quantification, and analysis to help future prevention and protection of global health, the economy, and ecosystems.

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