Development and application of a triplex‐PCR assay for rapid detection of methicillin‐resistant Staphylococcus aureus from pigs

Autor: G.P. Patil, Ravi Kant Agrawal, S R Pegu, Swaraj Rajkhowa
Rok vydání: 2020
Předmět:
Methicillin-Resistant Staphylococcus aureus
0106 biological sciences
Swine
Biology
medicine.disease_cause
01 natural sciences
Applied Microbiology and Biotechnology
Rapid detection
Microbiology
Methicillin
03 medical and health sciences
chemistry.chemical_compound
Bacterial Proteins
Limit of Detection
RNA
Ribosomal
16S
010608 biotechnology
medicine
Animals
Humans
Micrococcal Nuclease
Penicillin-Binding Proteins
DNA Primers
Detection limit
0303 health sciences
030306 microbiology
Staphylococcal Infections
biochemical phenomena
metabolism
and nutrition
bacterial infections and mycoses
16S ribosomal RNA
biology.organism_classification
Methicillin-resistant Staphylococcus aureus
chemistry
Staphylococcus aureus
Agarose
Methicillin Resistance
Multiplex Polymerase Chain Reaction
Triplex PCR
Bacteria
Zdroj: Letters in Applied Microbiology. 72:121-125
ISSN: 1472-765X
0266-8254
DOI: 10.1111/lam.13408
Popis: A triplex-PCR assay was developed and evaluated for rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) recovered from various biological samples of pig. Three sets of primers were designed to target mecA, 16S rRNA and nuc genes of MRSA. The specific amplification generated three bands on agarose gel, with sizes 280 bp for mecA, 654 bp for 16S rRNA and 481 bp for nuc, respectively. A potential advantage of the PCR assay is its sensitivity with a detection limit of 102 CFU per ml of bacteria. In all, 79 MRSA isolates recovered from various samples of pigs were subjected to the amplification by the triplex-PCR assay and all the isolates yielded three bands corresponding to the three genes under this study. No false-positive amplification was observed, indicating the high specificity of the developed triplex-PCR assay. This assay will be a useful and powerful method for differentiation of MRSA from methicillin-sensitive S. aureus, coagulase-negative methicillin-resistant staphylococci and coagulase-negative methicillin-sensitive staphylococci.
Databáze: OpenAIRE
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