scholarly journals Probing the dissonance among the diagnostic outputs of multiple approaches used for detection of Methicillin-resistant Staphylococcus aureus (MRSA)

2020 ◽  
Author(s):  
Ujjwal Ranjan Dahiya ◽  
Arnab Sikidar ◽  
Priyanka Sharma ◽  
Chitra Rawat ◽  
Benu Dhawan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Culture Method ◽  
Nucleic Acid Amplification ◽  
Detection Methods ◽  
Diagnostic Efficiency ◽  
Coagulase Negative Staphylococci ◽  
Culture Methods ◽  
Methicillin Resistant ◽  
Conventional Culture

Methicillin-resistant staphylococcus aureus (MRSA) is an extremely infectious hospital acquired bacterial pathogen often found in post-surgical patients globally. Early detection of such pathogens is a critical requirement to eliminate or reduce the incidence of antimicrobial resistance as well as for effective management of the disease. Despite the development of multiple biochemical, microbiological and nucleic acid amplification techniques (NAATs), conventional culture methods are widely used clinically owing to high variability between the methods, technical skills, and infrastructural needs. Further, multiple reports suggest a significant variation among diagnostic output for MRSA detection. This work attempts to probe the discordance among the diagnostic output of three commonly used methods while trying to understand the underlying cause of variability. MRSA detection on 217 clinical pus isolates was carried out using three different methods namely, conventional culture method, qPCR-based amplification, and a modern LAMP-based detection approach. Also, to confirm the presence of MRSA and distinguish from coagulase-negative staphylococci (CoNS), as well as to investigate the observed differences between qPCR and LAMP outputs, melt curve analysis was performed on discordant samples. LAMP-based MRSA detection was found to be the optimum method. In summary, this study evaluates the diagnostic efficiency of the different detection methods, while probing for possible explanations for the observed differences.

scholarly journals MecA gene and methicillin-resistant Staphylococcus aureus (MRSA) isolated from dairy farms in East Java, Indonesia

2020 ◽  
Vol 21 (8) ◽  
Author(s):  
Sancaka Chasyer Ramandinianto ◽  
Aswin Rafif Khairullah ◽  
Mustofa Helmi Effendi
Keyword(s):  
Staphylococcus Aureus ◽  
Pathogenic Bacteria ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Dairy Farms ◽  
Detection Methods ◽  
Disc Diffusion ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Oxacillin Resistance

Abstract. Ramandinianto SC, Khairullah AR, Effendi MH. 2020. MecA gene and methicillin-resistant Staphylococcus aureus (MRSA) isolated from dairy farms in East Java, Indonesia. Biodiversitas 21: 3562-3568. Milk Borne Disease (MBD) can be caused by a variety of pathogenic bacteria, one of which is Staphylococcus aureus which has a large impact on aspects of public health. The therapy used to treat staphylococcal infection is Oxacillin preparations that can inhibit bacterial wall synthesis, but the adaptation of the mecA gene to staphylococcal cassette chromosome mec (SCCmec) causes the emergence of strains of methicillin-resistant S. aureus (MRSA). The purpose of this study was to detect the level of MRSA strain contamination in dairy cows in East Java by comparing the mecA gene, Oxacillin, and Cefoxitin Disc Diffusion Methods and Oxacillin Resistance Screen Agar (ORSA) detection methods. A total of 150 cow's milk samples were taken at 3 village dairy farms in East Java, samples were added to the enrichment media Buffer Pepton Water (BPW) and then isolates were planted and purified using Mannitol Salt Agar (MSA). The detection of MRSA was carried out by the Kirby Bauer disc diffusion preparation Cefoxitin 30 μg and Oxacillin 30 μg then confirmed by ORSA and the presence of mecA gene by the polymerase chain reaction (PCR) method. The results showed that from a total of 92 S. aureus isolates using Oxacillin disc test, 24 resistant isolates were obtained, using Cefoxitin disc test, 17 isolates were obtained, and using the ORSA test 18 MRSA isolates were obtained. MRSA isolates tested by PCR obtained evidence of 2 isolates of mecA gene. It can be concluded that the Oxacillin disc test was the highest sensitivity for detecting MRSA strain isolate, however, mecA gene was the golden standard to detect MRSA on the dairy farms.

Evaluation of Detection Methods for Screening Meat and Poultry Products for the Presence of Foodborne Pathogens

2005 ◽  
Vol 68 (12) ◽  
pp. 2637-2647 ◽  
Author(s):  
VALERIE M. BOHAYCHUK ◽  
GARY E. GENSLER ◽  
ROBIN K. KING ◽  
JOHN T. WU ◽  
LYNN M. McMULLEN
Keyword(s):  
Foodborne Pathogens ◽  
Culture Method ◽  
Lateral Flow ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Processed Meat ◽  
Culture Methods ◽  
E Coli ◽  
Poultry Products ◽  
Conventional Culture

Rapid and molecular technologies such as enzyme-linked immunosorbent assay (ELISA), PCR, and lateral flow immunoprecipitation can reduce the time and labor involved in screening food products for the presence of pathogens. These technologies were compared with conventional culture methodology for the detection of Salmonella, Campylobacter, Listeria, and Escherichia coli O157:H7 inoculated in raw and processed meat and poultry products. Recommended protocols were modified so that the same enrichment broths used in the culture methods were also used in the ELISA, PCR, and lateral flow immunoprecipitation assays. The percent agreement between the rapid technologies and culture methods ranged from 80 to 100% depending on the pathogen detected and the method used. ELISA, PCR, and lateral flow immunoprecipitation all performed well, with no statistical difference, compared with the culture method for the detection of E. coli O157:H7. ELISA performed better for the detection of Salmonella, with sensitivity and specificity rates of 100%. PCR performed better for the detection of Campylobacter jejuni, with 100% agreement to the culture method. PCR was highly sensitive for the detection of all the foodborne pathogens tested except Listeria monocytogenes. Although the lateral flow immunoprecipitation tests were statistically different from the culture methods for Salmonella and Listeria because of false-positive results, the tests did not produce any false negatives, indicating that this method would be suitable for screening meat and poultry products for these pathogens.

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