Comparison of complete rpoB gene sequence typing and multi-locus sequence typing for phylogenetic analysis of Staphylococcus aureus

Seong, Won-Jin; Kim, Jae Hong; Kwon, Hyuk‐Joon

doi:10.2323/jgam.59.335

Cited by 9 publications

(16 citation statements)

References 27 publications

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“…A total of 84 S. aureus isolates from bovine mastitis cows (22 farms, 72 cows, and 84 udders) in Korea were isolated after cultivation on sheep blood agar and identified by using the VITEK2 system and Gram-Positive Identification Cards (BioMérieux, France) [ 18 ]. Fourteen human MRSA isolates were purchased from the Culture Collection of Antimicrobial Resistant Microbes (Korea), and MLST and rpoB sequence type (RST) were characterized previously [ 33 ] ( Supplementary Table 1 ). The 166 human strains were collected from the genome sequences of S. aureus in the GenBank database ( Supplementary Table 1 ).…”

Section: Methodsmentioning

confidence: 99%

“…The primer sets for RSTing and detection of the methicillin-resistance gene, mecA , and spa typing were used as previously reported [ 19 33 38 ]. The primer sets for mPPTing were designed by using large subunit terminase genes of S. aureus collected from the GenBank database; primer sequences are summarized in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

“…1.5; Technelysium, Australia). After comparison with consensus and previously identified RST sequences, the RST of the new rpoB sequence was assigned as previously described [ 33 ]. Nucleotide similarity, variable nucleotide comparisons, and translation of nucleotide sequences were performed with Bioedit software (ver.…”

Section: Methodsmentioning

confidence: 99%

“…Although comparative genomics has been successfully applied to elucidate the transmission and evolution of whole genetic contents related to antibiotic resistance and pathogenicity, it remains costly and time-consuming when analyzing large datasets. Sub-genomic methods including multi-locus sequence typing (MLST), arbitrary primer-polymerase chain reaction (PCR), pulsed-field gel electrophoresis, multi-locus variable number tandem repeat analysis, spa typing, small genomic islet typing, rpoB sequence typing (RSTing), and microarray hybridization have been used [ 5 8 11 26 27 28 33 41 ]. Among these methods, RSTing has been developed to reduce the experimental cost of MLST and was successfully applied to the molecular epidemiological study of human and avian isolates of S. aureus [ 33 ].…”

Section: Introductionmentioning

confidence: 99%

“…Sub-genomic methods including multi-locus sequence typing (MLST), arbitrary primer-polymerase chain reaction (PCR), pulsed-field gel electrophoresis, multi-locus variable number tandem repeat analysis, spa typing, small genomic islet typing, rpoB sequence typing (RSTing), and microarray hybridization have been used [ 5 8 11 26 27 28 33 41 ]. Among these methods, RSTing has been developed to reduce the experimental cost of MLST and was successfully applied to the molecular epidemiological study of human and avian isolates of S. aureus [ 33 ]. Furthermore, RSTing provides information on rifampin, daptomycin, and vancomycin resistance in S. aureus , which are useful for MRSA treatment [ 1 6 45 46 ].…”

Section: Introductionmentioning

confidence: 99%

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Molecular prophage typing ofStaphylococcus aureusisolates from bovine mastitis

Seong

Kim

et al. 2018

J Vet Sci

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Staphylococcus aureus is one of the major pathogens causing bovine mastitis and foodborne diseases associated with dairy products. To determine the genetic relationships between human and bovine or bovine isolates of S. aureus, various molecular methods have been used. Previously we developed an rpoB sequence typing (RSTing) method for molecular differentiation of S. aureus isolates and identification of RpoB-related antibiotic resistance. In this study, we performed spa typing and RSTing with 84 isolates from mastitic cows (22 farms, 72 cows, and 84 udders) and developed a molecular prophage typing (mPPTing) method for molecular epidemiological analysis of bovine mastitis. To compare the results, human isolates from patients (n = 14) and GenBank (n = 166) were used for real and in silico RSTing and mPPTing, respectively. Based on the results, RST10-2 and RST4-1 were the most common rpoB sequence types (RSTs) in cows and humans, respectively, and most isolates from cows and humans clearly differed. Antibiotic resistance-related RSTs were not detected in the cow isolates. A single dominant prophage type and gradual evolution through prophage acquisition were apparent in most of the tested farms. Thus, RSTing and mPPTing are informative, simple, and economic methods for molecular epidemiological analysis of S. aureus infections.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecular prophage typing ofStaphylococcus aureusisolates from bovine mastitis

Seong

Kim

et al. 2018

J Vet Sci

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Evolution of a major bovine mastitic genotype (rpoB sequence type 10-2) of Staphylococcus aureus in cows

Kim

et al. 2019

J Microbiol.

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Evolution of a major bovine mastitic genotype (rpoB sequence type 10-2) of Staphylococcus aureus in cows §Staphylococcus aureus is the major pathogen leading to bovine mastitis globally while livestock-associated methicillin resistant S. aureus (LA-MRSA) has become a potential threat to public health. MRSA from bovine mastitis is not common but a methicillin susceptible S. aureus (MSSA) genotype, rpoB sequence type (RST)10-2 (RST10-2), is prevalent in Korea. To date, many genomic sequences from S. aureus have been elucidated, but the complete genome sequences of RST10-2 MSSA from bovine mastitis has never been reported. In this study, we determined the complete genome sequence of two RST10-2 MSSA that differ from each other in staphylococcal protein A and molecular prophage types [PMB64-1 (t2489/ mPPT0) and PMB81-4 (t127/mPPT1-2-3)] and conducted a comparative genomics study. The genomic sequences of PMB64-1 and PMB81-4 were more homologous to the representative human RST10-2 strains (MSSA476, MW2 etc.) compared to other RSTs. Most of them shared five common pseudogenes, along with high amino acid identity of four variable virulence genes that were identified in this study. However, PMB64-1 and PMB81-4 acquired different strainspecific pseudogenes and mobile genetic elements than the human strains. The unique pseudogene profile and high identity of the virulence genes were verified in RST10-2 field strains from bovine mastitis. Thus, bovine mastitic RST10-2 MSSA may have an evolutionary relationship with the human RST10-2 community-associated (CA) MSSA and CA-MRSA strains but may have adapted to cows.

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