Coagulase-negative Staphylococci Species Research Articles

COAGULASE NEGATIVE STAPHYLOCOCCUS SPECIES

Objective: Department and sample wise distribution along with drug-resistantpattern and best therapeutic choice of drugs against Coagulase-negative Staphylococcusspecies (CoNS). Study Design: Cross-sectional study. Setting: Department of MicrobiologyAIMC Lahore Pakistan. Period: 1st January 2016 to 25th May 2017. Methodology: About4597 samples were collected from various departments and processed for antimicrobialresistant testing. Results: Of 4597 samples, culture positive were 40.4% CoNS were 9.0%and 22.3% of total and total culture positive specimens respectively The highest rate isolationrate of CONS was found in ICU 32.4%, Sample-wise 34.1% in blood samples.0% resistantto Linezolid, Teicoplanin, and Vancomycin, while very high resistant rate against penicillin77.8%, Cotrimoxazole 60%, Methicillin 59.8%,Co-amoxiclav 54%,Clindamycin 48.7%,FusidicAcid 463%, Doxycycline 45.1%, Erythromycin 44.9%, Amikacin 42%, Ciprofloxacin 41.8% andGentamycin 23%. Conclusion: Linezolid, Vancomycin, and Teicoplanin are the best choice ofdrugs while emergence of drug-resistant to other basic drugs is alarming

Complete Genome Sequence of Staphylococcus haemolyticus Type Strain SGAir0252.

ABSTRACTStaphylococcus haemolyticus is a coagulase-negative staphylococcal species that is part of the skin microbiome and an opportunistic human pathogen. The strain SGAir0252 was isolated from tropical air samples collected in Singapore, and its complete genome comprises one chromosome of 2.63 Mb and one plasmid of 41.6 kb.

RESISTANCE AND THERAPEUTIC DECISIONS AT THE TURN OF THE NOVEL MILLENNIUM

Objective: Department and sample wise distribution along with drug-resistantpattern and best therapeutic choice of drugs against Coagulase-negative Staphylococcusspecies (CoNS). Study Design: Cross-sectional study. Setting: Department of MicrobiologyAIMC Lahore Pakistan. Period: 1st January 2016 to 25th May 2017. Methodology: About4597 samples were collected from various departments and processed for antimicrobialresistant testing. Results: Of 4597 samples, culture positive were 40.4% CoNS were 9.0%and 22.3% of total and total culture positive specimens respectively The highest rate isolationrate of CONS was found in ICU 32.4%, Sample-wise 34.1% in blood samples.0% resistantto Linezolid, Teicoplanin, and Vancomycin, while very high resistant rate against penicillin77.8%, Cotrimoxazole 60%, Methicillin 59.8%,Co-amoxiclav 54%,Clindamycin 48.7%,FusidicAcid 463%, Doxycycline 45.1%, Erythromycin 44.9%, Amikacin 42%, Ciprofloxacin 41.8% andGentamycin 23%. Conclusion: Linezolid, Vancomycin, and Teicoplanin are the best choice ofdrugs while emergence of drug-resistant to other basic drugs is alarming

Simple and economical method for identification and speciation of Staphylococcus epidermidis and other coagulase negative Staphylococci and its validation by molecular methods

Coagulase-negative staphylococci (CoNS) have been increasingly recognized as a clinically important group of species that can cause several opportunistic nosocomial infections. There are at least 47 known species of Staphylococci and to differentiate all these species >40 biochemical tests need to be performed. The present study was able to refine the CoNS identification process by using only five tests to identify S. epidermidis from the rest and used six other tests to identify eleven other clinically significant CoNS species. A total of 242 CoNS isolates were collected from tertiary care hospitals and included in the study. The five-biochemical test scheme devised based on mathematical probability derived from a computer algorithm included fermentation of mannitol, maltose, mannose, trehalose and novobiocin susceptibility to differentiate S. epidermidis from other CoNS species. The remaining CoNS isolates other than S. epidermidis were further characterized with the help of six additional tests, which identified another eleven species. Species-specific PCR and 16SrDNA sequencing were used to confirm and validate the identification scheme. Species-specific PCR and 16SrDNA sequencing showed 100% agreement with non-divergent phenotypic test results, indicating that the five selected assays are highly specific for identifying S. epidermidis. In conclusion, this study used only 11 tests to identify most of the clinically significant CoNS that can reduce cost and time. This scheme is easy to perform in any laboratory with basic resources, the results of this study were validated using more accurate molecular methods such as PCR and 16S rDNA typing to confirm the utility of the proposed scheme.

Haemato-Biochemical Changes in Staphylococcal Species Induced Mastitis in Mouse Model

In this study, haemato-biochemical changes in mouse mastitis by inoculating Staphylococcus epidermidis, S. chromogenes, S. haemolyticus and S. Aureus isolated from bovine milk was studied. Blood was collected by intracardiac route at 6, 12, 24, 48, 72 and 96 h. Haematological changes noticed were: white blood cells (WBC) decreased in S. haemolyticus and S. chromogenes at 48 h; S. haemolyticus and S. aureus at 96 h. S. haemolyticus showed increased lymphocyte at 24 and 48 h and in S. aureus at 48 h. Neutrophil was decreased in S. haemolyticus and S. aureus group. Glucose increased at 6 h with three Coagulase Negative Staphylococcus (CNS) species and S. aureus. Glucose and total protein increased in S. chromogenes infected mice. Globulin levels showed decrease in S. epidermidis and S. aureus. Albumin globulin ratio showed decreasing trend in S. chromogenes and S. haemolyticus inoculated mice. Aspartate aminotransferase (AST) level increased at 6 and 12 h in all infected groups. Alanine aminotransferase increased at 12 and 48 h in S. chromogenes and S. aureus respectively. Lactate dehydrogenase (LDH) increased at 6 h in all the infected mice. Thus, three CNS species induced haemato-biochemical changes in haemoglobin, WBC, neutrophil, glucose, total protein, AST and LDH which may be used as indicators for diagnosis of CNS species mastitis in mice. Keywords: Staphylococcus species, mouse mastitis, haemato-biochemical changes Cite this Article Krishnamoorthy P, Satyanarayana ML, Shome BR, et al. Haemato-Biochemical Changes in Staphylococcal Species induced Mastitis in Mouse Model + . Research & Reviews: Journal of Veterinary Science and Technology . 2015; 4(3): 8–14p.

Elimination of experimentally induced bovine intramammary infection assessed by multiplex real-time PCR and bacterial culture

Diagnosis of bovine intramammary infection (IMI) has traditionally been based on bacterial culture, but currently IMI can also be detected with DNA based methods, such as multiplex real-time PCR. The aim of this study was to describe the elimination of bacteria in experimentally induced IMI on the quarter level, using conventional bacterial culture (BC) and multiplex real-time PCR. Two coagulase-negative staphylococcal species, Staphylococcus epidermidis and Staphylococcus simulans, were experimentally inoculated into 14 healthy quarters of 8 dairy cows during 4 consecutive study periods. Intramammary infections were followed with 20 milk samplings per each quarter. Milk somatic cell count was monitored to evaluate the inflammation process in the quarters. Four quarters cured spontaneously during the study period according to the culture. The PCR detected staphylococcal DNA from these quarters for several days after they were defined as cured in BC. Agreement between BC and PCR results varied from substantial to almost perfect agreement for the first 36 h postchallenge, decreasing to moderate levels toward the end of the sampling period. Based on this study, we recommend collecting possible follow-up samples to assess the bacteriological cure from IMI not until 2 to 3 wk after the onset of mastitis or after the quarter milk somatic cell count has normalized when PCR is used.

The narrowing down of inoculated communities of coagulase-negative staphylococci in fermented meat models is modulated by temperature and pH

Coagulase-negative staphylococci (CNS) are involved in colour and flavour formation of fermented meats. Their communities are established either spontaneously, as in some artisan-type products, or using a starter culture. The latter usually consists of Staphylococcus carnosus and/or Staphylococcus xylosus strains, although strains from other CNS species also have potential for application. However, it is not entirely clear how the fitness of alternative starter cultures within a fermented meat matrix compares to conventional ones and how this may be affected by processing conditions. Therefore, the aim of this study was to assess the influence of two key processing conditions, namely temperature and acidity, on the competitiveness of a cocktail of five different strains of CNS belonging to species that are potentially important for meat fermentation (Staphylococcus xylosus 2S7-2, S. carnosus 833, Staphylococcus epidermidis ATCC 12228, Staphylococcus equorum DFL-S19, and Staphylococcus saprophyticus FPS1). To this end, fermented meat models consisting of cured meat batters with initial pH values of 5.3, 5.5, or 5.7 were inoculated with these strains, stuffed in containers, and incubated at 23, 30, or 37 °C. Both the pH level and the temperature influenced the composition of the CNS communities, giving a competitive advantage to the best adapted species. Staphylococcus xylosus preferred low temperature and mild acidity, whereas an elevated temperature selected for S. epidermidis and a low pH for S. carnosus. Under the conditions tested, S. saprophyticus and S. equorum were outcompeted by the three other CNS species. Hence, CNS communities in fermented meats are not only established based on the initial presence of specific species in the meat batter but also by their subsequent adaptation to the processing conditions during fermentation, potentially overruling the use of starter cultures.

Small mammals as sentinels of antimicrobial-resistant staphylococci.

A total of 39 coagulase-negative staphylococci and seven Staphylococcus aureus strains were isolated from small mammal feces, i.e., the striped field mouse (Apodemus agrarius) and the yellow-necked mouse (A. flavicollis) in two sampling areas, deciduous forest and karst plains. MALDI-TOF analysis revealed five species of coagulase-negative staphylococci: S. sciuri, S. hominis, S. warneri, S. haemolyticus, and S. xylosus. All strains were susceptible to tetracycline, linezolid, vancomycin, and teicoplanin. Three MRSA strains with the mecA gene were detected. The beta-lactamase gene blaZ was detected in ampicillin-resistant staphylococci and in the high-level resistant strains (oxacillin over 2mg/L) mecA gene. The mecC gene was not detected by PCR. Erythromycin-resistant staphylococci harbored the ermC gene and/or the efflux gene msrA. There were no detectable dfr genes in trimethoprim-resistant staphylococci and the rifampicin-resistant strains were without mutation in the rpoB gene. In summary, wild small mammals may serve as sentinels of mecA-positive S. aureus with erythromycin resistance genes ermC and efflux msrA. Small mammals appear to be useful indicators of antibiotic resistance.

Characterization of Coagulase-Negative Staphylococci and pheno-genotypic beta lactam resistance evaluation in samples from bovine Intramammary infection

ABSTRACT This study aimed to identify Coagulase-Negative Staphylococci (CoNS) species isolated from bovine mastitis, through phenotypic and PCR-RFLP (Restriction Fragment Length Polymorphism-Polimerase Chain Reaction) methods and to compare both techniques to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technique. Among them, the PCR-RFLP method, using a partially conserved sequence of the groEL gene, is a promising alternative, because of its reproducibility and reliability. On the other hand, the proteomic technique MALDI-TOF MS provides an accurate and much faster diagnosis and has been increasingly employed in microbiological identification. The pheno-genotypic profiles of beta-lactam resistance were also investigated, this characterization is important, considering that the use of antimicrobials is a key element for mastitis control in dairy farms. The concordance of the phenotypic, PCR-RFLP and MALDI-TOF MS assays to identify CoNS species was 77,5% (31/40). S. chromogenes was the species most frequently isolated. Antibiotic resistance rate was relatively low, registering values of 25.5% to penicillin, 9.6% to oxacillin and 6.2% to cefoxitin. Resistance to imipenem, cephalotin and amoxicillin+clavulanate was not observed. The mecA gene and its variant were detected in 7.6% and 4,1% of the isolates respectively. The blaZ gene was found in 43.2% of the strains resistant to penicillin.

Pathogenesis and inflammatory response in experimental caprine mastitis due to Staphylococcus chromogenes

Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria in cases of subclinical mastitis in dairy cows. CNS species may differ in their pathogenicity, but very little is known about their virulence factors or their immune response in intramammary infections. To our knowledge, no experimental studies into the mastitis pathogenesis caused by CNS have been described in lactating goats. The aim of this study was to induce an experimentally Staphylococcus chromogenes mastitis in lactating goats aimed at verifying if the model can be used to evaluate the inflammatory response, the dynamics of infection and the pathological findings within the first hours of intramammary inoculation. Six Saanen goats in mid-lactation were inoculated with 1 × 107 colony forming units of S. chromogenes. Bacterial growth peaked in milk from the challenged right halves of the mammary glands (RMG) at 4 h post inoculation (PI). Shedding of viable bacteria showed a marked decrease at 12 h PI. An increase in mean somatic cell counts was observed in the milk samples from 8 h PI onwards. Mild clinical signs were evoked by intramammary inoculation. Staphylococcus chromogenes could be isolated in tissue from all RMG. Histological examination of specimens of the RMG and lymph nodes of the goats showed an increased inflammatory response throughout the experiment with respect to control halves. In conclusion, the experimental inoculation of S. chromogenes in lactating goats is capable of eliciting an inflammatory response and capable of causing pathological changes. This research represents a preliminary study for a better knowledge of the mastitis pathogenesis caused by S. chromogenes.

Matrix-assisted laser desorption ionization-time of flight: a promising alternative method of identifying the major coagulase-negative Staphylococci species.

This study aimed to evaluate different methods for differentiation of species of coagulase-negative staphylococci (CoNS) that caused infections in hospitalized immunocompromised patients. A total of 134 CoNS strains were characterized using four different methods. The results of matrix assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) analysis were in complete agreement with those of tuf gene sequencing (kappa index = 1.00). The kappa index of Vitek 2® Compact analysis was 0.85 (very good) and that of the conventional method was 0.63 (moderate). . MALDI-TOF MS provided rapid and accurate results for the identification of CoNS (134; 100%).

Phenotypic and genotypic investigation of methicillin resistant Staphylococci species isolated from children with sepsis in Egypt

S TAPHYLOCOCCUS spp. has emerged as a major cause of sepsis. Methicillin resistance is a pattern of resistance which hinders the management of such infection. The aims of the present study were to isolate Staphylococci spp. from children with clinical signs and symptoms of sepsis at Mansoura University Children Hospital (MUCH), Egypt. Then to detect methicillin resistance Staphylococci (MRS) using primary screening test followed by molecular typing of the methicillin resistant Staphylococci isolates by multiplex PCR. MRS sepsis was diagnosed in a total 100 patient from February 2015, till February, 2016. 18 (18%) of isolates were identified as S. aureus and 82 (82%) of Staphylococci spp. were identified as coagulase negative Staphylococci (CoNS). S. hominis subsp. hominis was the commonest species of CoNS (32/ 82, 39%), followed by S. sciuri (9/82, 11%). the rate of Oxacillin resistance among all isolated Staphylococci strains was (100%) in both S. aureus and CoNS isolates.Among the 50 selected resistance species for multiplex PCR there 27 (54%) species had a single gene SCCmec type I or SCCmec type (II or III). While 13 (26%) isolates had both two genes (SCCmec type I + II or III) and 10 (2%) had no SCCmec genes. The spectrum of children with sepsis at Mansoura University Children Hospital (MUCH), confirmed the importance of pathogens such as Staphylococcus spp. Similar to other studies from the developing countries, CoNS species were the most common isolated pathogens in children with sepsis.

Pathogenicity and antibiotic resistance of coagulase-negative staphylococci isolated from retailing chicken meat

Diverse coagulase-negative staphylococci (CNS) species isolated from meat are previously reported and considered as potential pathogenic staphylococci through obtain horizontal transferred elements (e.g. toxic, antibiotic resistance factors) from other bacteria. In this study, prevalence, homology and pathogenicity of CNS isolated from 180 raw chicken meat samples in retail markets were studied. The overall prevalence of CNS was 11.7% and centralized in 69.2% of Staphylococcus epidermidis and 30.8% of Staphylococcus warneri among diverse sources (including different retail settings, suppliers and products). Molecular typing showed highly homogenous patterns in the same species. Antibiotic susceptibility test revealed a common penicillin and erythromycin resistance among all of CNS, while 17 S. epidermidis isolates displayed multidrug resistance of gentamycin (GM)-kanamycin (K)-penicillin (P)-erythromycin (E), except one methicillin-resistant S. epidermidis isolate which was typed in staphylococcal cassette chromosome mec (SCCmec) Ⅴ. Conclusively, diverse sourced raw chicken meat products exhibited a centralized prevalence of S. epidermidis and S. warneri, and a highly homogenous of genome in the same CNS specie. Though the negative toxigenicity of CNS was determined, constant multidrug resistance in S. epidermidis and P-E resistance in CNS need to be highly concerned regarding human health.

Coagulase-Negative Staphylococcal Strain Prevents Staphylococcus aureus Colonization and Skin Infection by Blocking Quorum Sensing

Coagulase-negative staphylococci (CoNS) and Staphylococcus aureus are part of the natural flora of humans and other mammals. We found that spentmedia from the CoNS species Staphylococcus caprae can inhibit agr-mediated quorum sensing by allclasses of S.aureus. A biochemical assessmentofthe inhibitory activity suggested that the S. caprae autoinducing peptide (AIP) was responsible,and mass spectrometric analysis identified the S.caprae AIP as an eight-residue peptide (YSTCSYYF). Using a murine model of intradermal MRSA infection, the therapeutic efficacy of synthetic S.caprae AIP was evident by a dramatic reduction in both dermonecrotic injury and cutaneous bacterial burden relative to controls. Competition experiments between S.caprae and MRSA demonstrated a significant reduction in MRSA burden using murine models of both skin colonization and intradermal infection. Our findings indicate that important interactions occur between commensals that can impact disease outcomes and potentially shape the composition of the natural flora.

PERSISTÊNCIA DE STAPHYLOCOCCUS COAGULASE-NEGATIVOS EM GLÂNDULAS MAMÁRIAS DE OVELHAS COM MASTITE SUBCLÍNICA APÓS O TRATAMENTO ANTIMICROBIANO À SECAGEM

Resumo Staphylococcus coagulase-negativos (SCN) são os principais micro-organismos responsáveis pela mastite ovina e o tratamento ao final da lactação pode ser usado como método de controle contra a doença. Porém, o longo período seco apresentado pelas ovelhas em alguns sistemas de criação pode prejudicar os efeitos positivos do tratamento antimicrobiano. Os objetivos deste trabalho foram identificar as principais espécies de SCN na etiologia da mastite ovina antes e após o tratamento ao final da lactação das ovelhas, bem como investigar a persistência das espécies mais prevalentes na glândula mamária na lactação seguinte. Sessenta ovelhas foram divididas em dois grupos experimentais, um deles formado por animais sem tratamento antimicrobiano, enquanto o outro era composto por ovelhas cujas metades mamárias foram tratadas com cloxacilina-benzatina por via intramamária. As amostras de leite foram obtidas antes da secagem e aos 15 e 30 dias após o parto da lactação seguinte. As espécies prevalentes foram S. warneri, S. simulans e S. epidermidis. Clones das três espécies de maior ocorrência foram identificados antes e depois do tratamento, ou seja, mesmo com o extenso período seco entre as duas lactações consecutivas, os micro-organismos ainda foram identificados no interior da glândula mamária.

Short communication: Evaluation of MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci

This study evaluated MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci (CNS). A total of 861 CNS isolates were used in the study, covering 21 different CNS species. The majority of the isolates were previously identified by rpoB gene sequencing (n = 804) and the remainder were identified by sequencing of hsp60 (n = 56) and tuf (n = 1). The genotypic identification was considered the gold standard identification. Using a direct transfer protocol and the existing commercial database, MALDI-TOF mass spectrometry showed a typeability of 96.5% (831/861) and an accuracy of 99.2% (824/831). Using a custom reference spectra expanded database, which included an additional 13 in-house created reference spectra, isolates were identified by MALDI-TOF mass spectrometry with 99.2% (854/861) typeability and 99.4% (849/854) accuracy. Overall, MALDI-TOF mass spectrometry using the direct transfer method was shown to be a highly reliable tool for the identification of bovine-associated CNS.

원유에서 Coagulase-Negative Staphylococci 검출율 및 특징

Mastitis is a common and serious infection of the mammary gland in dairy cattle and has a major economic impact on the production of milk and dairy products. Bacterial mastitis is caused by several pathogens and is most frequently associated with coagulase-negative staphylococci (CNS). Although CNS are typically associated with subclinical or mild mastitis, the importance of CNS has increased as these pathogens have emerged as predominant mastitis-related pathogens in many countries. CNS can cause persistent infections, resulting in increased milk somatic cell counts and thereby affecting milk quality and decreasing milk production. Globally, Staphylococcus chromogenes, S. epidermidis, and S. simulans are the predominant CNS species in dairy cattle mastitis. Antibacterial resistance of CNS varies with species, and most CNS are susceptible to vancomycin and resistant to penicillin and tetracycline. As the most frequently isolated CNS species, some strains of S. chromogenes exhibit phenotypic resistance to ampicillin, erythromycin, oxacillin, penicillin, and tetracycline. Some strains of S. epidermidis and S. haemolyticus are only susceptible to vancomycin and rifampicin. Therefore, more studies are needed to achieve the control and prevention of CNS as environmental pathogens.

INTRAVENOUS LINE COLONIZATION IN PATIENTS RECEIVING CYTOTOXIC DRUGS

Background
 Hospital acquired infections are infections that develop during the hospital stay; they mainly include urinary tract, respiratory tract or blood stream infections. 
 Objectives
 To investigate microbial colonization of intravenous cannula of cancers patients receiving cytotoxic drugs. 
 Materials and Methods
 Colonization of intravenous cannulas was investigated by cultivation of the device surface and lumen on different culture media under aerobic incubation. The isolates were identified based on cultural, morphological and biochemical activities. The susceptibility of the isolates to antimicrobial drugs was investigated using Bauer-Kirby disk diffusion method and the bacterial isolates were further investigated for their ability to form biofilm. 
 Results
 From 200 cultivated intravenous cannulas, microbial colonization was detected among 26% and 11% of intravenous cannulas from cancer and non-cancer patients respectively; Out of 37 isolates, 34 (91.9 %) were Gram-positive bacteria, 2 (5.4 %) Gram-negative bacteria and one (2.7 %) was a Candida species. The main bacterial isolates were bacteria belonging to different coagulase negative staphylococci species such as Staphylococcus epidermidis, S. hominis and others. Few faecal organisms were isolated including a single isolate of each Klebsiella pneumoniae and Escherichia coli. All bacterial isolates were able to form biofilm and most showed to have biofilm-related icaA and icaD genes.
 Conclusion
 We detected microbial colonization of intravenous cannulas in 18.5% of all cultivated devices. We observed more colonization in cancer patients (26%) in comparison to the control group (11%). Most of the isolates were species of CoNS that were able to form biofilm.

CRISPR-Cas Systems Features and the Gene-Reservoir Role of Coagulase-Negative Staphylococci.

The claimed role of gene reservoir of coagulase-negative staphylococci (CoNS) could be contradicted by estimates that CRISPR/Cas systems are found in the genomes of 40–50% of bacteria, as these systems interfere with plasmid uptake in staphylococci. To further correlate this role with presence of CRISPR, we analyzed, by computational methods, 122 genomes from 15 species of CoNS. Only 15% of them harbored CRISPR/Cas systems, and this proportion was much lower for S. epidermidis and S. haemolyticus, the CoNS most frequently associated with opportunistic infections in humans. These systems are of type II or III, and at least two of them are located within SCCmec, a mobile genetic element of Staphylococcus bacterial species. An analysis of the spacers of these CRISPRs, which come from exogenous origin, allowed us to track the transference of the SCCmec, which was exchanged between different strains, species and hosts. Some of the spacers are derived from plasmids described in Staphylococcus species that are different from those in which the CRISPR are found, evidencing the attempt (and failure) of plasmid transference between them. Based on the polymorphisms of the cas1 gene in CRISPRs of types II and III, we developed a multiplex polymerase chain reaction (PCR) suitable to screen and type CRISPR systems in CoNS. The PCR was tested in 59 S. haemolyticus strains, of which only two contained a type III cas1. This gene was shown to be expressed in the exponential growth, stationary phase and during biofilm formation. The low abundance of CRISPRs in CoNS is in accordance with their role as gene reservoirs, but when present, their spacers sequence evidence and give an insight on the dynamics of horizontal genetic transfer among staphylococci.

Improved multiplex PCR primers for rapid identification of coagulase-negative staphylococci

Coagulase-negative staphylococci (CNS) are opportunistic pathogens that are currently emerging as causative agents of human disease. Though CNS are widespread in the clinic and food, their precise identification at species level is important. Here, using 16S rRNA sequencing, 55 staphylococcal isolates were identified as S. capitis, S. caprae, S. epidermidis, S. haemolyticus, S. pasteuri, S. saprophyticus, S. warneri, and S. xylosus. Although 16S rRNA sequencing is universally accepted as a standard for bacterial identification, the method did not effectively discriminate closely related species, and additional DNA sequencing was required. The divergence of the sodA gene sequence is higher than that of 16S rRNA. To devise a rapid and accurate identification method, sodA-specific primers were designed to demonstrate that species-specific multiplex polymerase chain reaction (PCR) can be used for the identification of CNS species. The accuracy of this method was higher than that of phenotypic identification; the method is simple and less time-consuming than 16S rRNA sequencing. Of the 55 CNS isolates, 92.72% were resistant to at least one antibiotic, and 60% were resistant to three or more antibiotics. CNS isolates produced diverse virulence-associated enzymes, including hemolysin (produced by 69.09% of the isolates), protease (65.45%), lipase (54.54%), lecithinase (36.36%), and DNase (29.09%); all isolates could form a biofilm. Because of the increasing pathogenic significance of CNS, the efficient multiplex PCR detection method developed in this study may contribute to studies for human health.