scholarly journals Rapid Detection of Staphylococcus aureus and Related Species Isolated from Food, Environment, Cosmetics, a Medical Device, and Clinical Samples Using the VITEK MS Microbial Identification System

2018 ◽  
Vol 101 (4) ◽  
pp. 1135-1143 ◽  
Author(s):  
Irshad M Sulaiman ◽  
Pratik Banerjee ◽  
Ying-Hsin Hsieh ◽  
Nancy Miranda ◽  
Steven Simpson ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Medical Device ◽  
Food Environment ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Identification System ◽  
Vitek 2 ◽  
Staphylococcus Spp ◽  
Vitek Ms

Abstract Staphylococcus spp. is considered as one of the most common human-pathogenic bacteria, causing illnesses ranging from nonthreatening skin infections to lethal diseases, including sepsis, pneumonia, bloodstream infections, and food poisoning. The emergence of methicillin-resistant Staphylococcus aureus strains has increased morbidity and mortality and resulted in a major healthcare burden worldwide. Single and multilocus sequence typing have been extensively used in the identification of Staphylococcus species. Nevertheless, these assays are relatively time-consuming and require high-quality DNA. Matrix-assisted laser desorption ionization-time-of-flight has been used recently for the rapid identification of several bacterial species. In this study, we have examined 47 Staphylococcus isolates recovered from food, environment, clinical samples, cosmetic products, and a medical device and 3 American Type Culture Collection Staphylococcus reference isolates using bioMérieux VITEK MS and VITEK 2 systems to determine isolate identity. Sequencing of the 16S ribosomal RNA gene was performed to confirm and compare the species identification data generated by VITEK 2 and VITEK MS systems. Although the VITEK 2 system could not identify one of the isolates, VITEK MS identified all 50 Staphylococcus spp. isolates tested. Results of this study clearly suggest that VITEK MS can be used in the rapid identification of Staphylococcus isolates of public health importance.

scholarly journals Comparison of PCR and phenotypic methods for the detection of methicillin resistant Staphylococcus aureus

2021 ◽  
Author(s):  
Anitha Madhavan ◽  
Arun Sachu ◽  
Anukumar Balakrishnan ◽  
Anu Vasudevan ◽  
Sobha Balakrishnan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Reference Method ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Disc Diffusion ◽  
Methicillin Resistant ◽  
Zone Diameter ◽  
Gold Standard Reference ◽  
Vitek 2 ◽  
Phenotypic Methods

Background and Objectives: Resistance to methicillin in methicillin resistant strains of Staphylococcus aureus (MRSA) is due to the presence of mec-A gene ,which encodes a low affinity penicillin binding protein (PBP)-2a or PBP2. Accurate and rapid identification of MRSA in clinical specimens is essential for timely decision on effective treatment. The aim of the study was to compare three different methods for detection of MRSA namely cefoxitin disc diffusion, CHROM agar MRSA and VITEK-2 susceptibility with PCR which is the gold standard reference method and to find the antibiotic susceptibility pattern of these isolates by VITEK-2. Materials and Methods: A Total of 100 non-duplicate S. aureus isolates were collected from different clinical samples among both outpatient and inpatients. Detection of MRSA among these isolates was done by cefoxitin disc diffusion, VITEK-2, CHROM agar MRSA and PCR. Results: The sensitivity and specificity of cefoxitin disc diffusion and Vitek was found to be 97.2% and 100%, while that of CHROM agar was found to be 100% and 78.6%. The overall prevalence of MRSA in our study by PCR was 72%. Conclusion: Based on the findings in our study, isolates which show cefoxitin zone diameter < 22 mm can be reported as MRSA. However, those isolates which have a zone diameter between 22-24 mm, should ideally be confirmed by PCR.

scholarly journals Rapid identification of health care–associated infections with an integrated fluorescence anisotropy system

2016 ◽  
Vol 2 (5) ◽  
pp. e1600300 ◽  
Author(s):  
Ki Soo Park ◽  
Chen-Han Huang ◽  
Kyungheon Lee ◽  
Yeong-Eun Yoo ◽  
Cesar M. Castro ◽  
...  
Keyword(s):  
Health Care ◽  
Fluorescence Anisotropy ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Integrated System ◽  
Clinical Samples ◽  
Drug Resistant ◽  
Polarization Anisotropy ◽  
Major Health ◽  
Health Care Associated Infections

Health care–associated infections (HAIs) and drug-resistant pathogens have become a major health care issue with millions of reported cases every year. Advanced diagnostics would allow clinicians to more quickly determine the most effective treatment, reduce the nonspecific use of broad-spectrum antimicrobials, and facilitate enrollment in new antibiotic treatments. We present a new integrated system, polarization anisotropy diagnostics (PAD), for rapid detection of HAI pathogens. The PAD uses changes of fluorescence anisotropy when detection probes recognize target bacterial nucleic acids. The technology is inherently robust against environmental noise and economically scalable for parallel measurements. The assay is fast (2 hours) and performed on-site in a single-tube format. When applied to clinical samples obtained from interventional procedures, the PAD determined the overall bacterial burden, differentiated HAI bacterial species, and identified drug resistance and virulence status. The PAD system holds promise as a powerful tool for near-patient, rapid HAI testing.

scholarly journals SCA Medium: A New Culture Medium for the Isolation of All Candida auris Clades

2021 ◽  
Vol 7 (6) ◽  
pp. 433
Author(s):  
Ahmad Ibrahim ◽  
Lucie Peyclit ◽  
Rim Abdallah ◽  
Saber Khelaifia ◽  
Amanda Chamieh ◽  
...  
Keyword(s):  
Culture Medium ◽  
Limit Of Detection ◽  
Bacterial Species ◽  
Multidrug Resistant ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Candida Auris ◽  
Phenotypic Profile ◽  
Stool Samples ◽  
Selective Culture Medium

Candida auris is an emerging multidrug-resistant yeast causing nosocomial infections and associated with high mortality in immunocompromised patients. Rapid identification and characterisation are necessary for diagnosis and containing its spread. In this study, we present a selective culture medium for all C. auris clades. This medium is sensitive with a limit of detection ranging between 101 and 102 CFU/mL. The 100% specificity of SCA (specific C. auris) medium is confirmed on a set of 135 Candida strains, 50 bacterial species and 200 human stool samples. Thus, this medium specifically selects for C. auris isolation from clinical samples, allowing the latter to study its phenotypic profile.

scholarly journals Phylogenetic tree analysis based on the 16S sequence alignment for Klebsiella spp. isolated from different sources

2019 ◽  
pp. 1957-1966
Author(s):  
Mustafa Basil Abdul Qader ◽  
Marwa Hameed AlKhafaji
Keyword(s):  
16S Rrna ◽  
Klebsiella Pneumoniae ◽  
16S Rrna Gene ◽  
Enteric Bacteria ◽  
Apium Graveolens ◽  
Clinical Samples ◽  
Identification System ◽  
Rrna Gene ◽  
Vitek 2 ◽  
Klebsiella Spp

16S ribosomal RNA (16S rRNA) gene sequences used to study bacterial phylogeny and taxonomy have been by far the most common housekeeping genetic marker utilized for identification and ancestor determination. This study aimed to investigate, for the first time, the relationship between Klebsiella spp. isolated from clinical and environmental samples in Iraq.      Fifty Klebsiella spp. isolates were isolated from clinical and environmental sources. Twenty-five isolates were collected from a fresh vegetable (Apium graveolens) and 25 from clinical samples (sputum, wound swab, urine). Enteric bacteria were isolated on selective and differential media and identified by an automatic identification system, vitek-2. The total DNA was extracted and PCR amplified for selected isolates. The 16S rRNA gene was amplified by using the universal primer 27F (5'- AGAGTTTGATCCTGGCTCAG- 3') and 1492R (5'- GGTTACCTTGTTACGACTT- 3’). The 16SrRNA gene sequence was analysed among some local isolates, and the results were compared with the standard data of similar registered strains in NCBI. The most common species of Klebsiella was Klebsiella pneumoniae pneumoniae (Kpp), followed by Klebsiella pneumoniae ozaenae (Kpo) and Klebsiella oxytoca (Ko). The results of the identification of species and sub species by using the  biochemical test (vitek-2) were more precise than those obtained by the use of the universal primer.Phylogenetic tree strategies have clearly indicated a relatively close similarity amongst all analysed Klebsiella isolates and revealed the intra-species genetic distance between the individual isolates of the Klebsiella spp. In conclusion, our results revealed the main advantage of using universal primers for the identification of Klebsiella spp. and their root from nature.

scholarly journals An Evaluation of Vitek MS System for Rapid Identification of Bacterial Species in Positive Blood Culture

2017 ◽  
Vol 49 (4) ◽  
pp. 407-412
Author(s):  
Kang-Gyun Park ◽  
Sang-Ha Kim ◽  
Jong-Tae Choi ◽  
Sunghyun Kim ◽  
Young-Kwon Kim ◽  
...  
Keyword(s):  
Blood Culture ◽  
Positive Blood Culture ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Vitek Ms

Rapid Identification of Methicillin-Resistant Staphylococcus aureus (MRSA) by the Vitek MS Saramis system

2015 ◽  
Vol 72 (1) ◽  
pp. 29-32 ◽  
Author(s):  
Weiguang Shan ◽  
Jiaping Li ◽  
Ying Fang ◽  
Xuan Wang ◽  
Danxia Gu ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Rapid Identification ◽  
Methicillin Resistant ◽  
Vitek Ms

scholarly journals Study the antibiotics sensitivity and beta- lactamase productivity of some Staphylococcus spp. Isolates from different sources of the Al Jamhoree Teaching Hospital in Mosul City

2019 ◽  
Vol 24 (7) ◽  
pp. 27
Author(s):  
Ayman Mohammed Khaleel1 ◽  
Adeeba Younis Shareef ◽  
, Shakeeba Younis Shareef
Keyword(s):  
Staphylococcus Aureus ◽  
Teaching Hospital ◽  
Clinical Laboratory ◽  
Bacterial Species ◽  
Age Groups ◽  
National Committee ◽  
Beta Lactamase ◽  
Bacterial Isolates ◽  
Bacterial Strains ◽  
Staphylococcus Spp

The study includes the isolation of certain types of Gram-positive bacteria Staphylococcus spp..The sample materials (blood, wounds, burns) were collected from both genders of all age groups of inpatients in Al Jamhoree Teaching Hospital in the Mosul during June till end of November 2018. Staphylococcus aureus was the most common among the isolates with 7 isolates(38.9%) followed by Staphylococcus epidermidis, Staphylococcus scriuri Staphylococcus lentus (Staphylococcus simulans) with 2 isolates each with 11.1% followed by Staphylococcus chromogenase, Staphylococcu scapilis and  Staphylococcus xylosus with one isolate for each one 5.5%. The number of isolates was 9 isolates and 50%, followed by wound samples. The isolates were 6 isolated by 33% and the samples of burns were 3 isolates. 17% were isolated and Staphylococcus aureus was the most dominant species. The sensitivity of bacterial isolates was  studied for 15 antibiotics for different antibiotic groups. The results showed that a difference in the resistance ratio of the isolates to these antagonists, as they were resistant to the Ceftrixone, Cloxacillin 100%. The Ciprofloxacillin, Gentamycin, Pipracillin, and Amikacin resistance were  reduced. The Ciprofloxacillin antagonist was the most affected on the bacterial isolates studied, followed by the Rifampin. The results showed that the beta-lactamase enzyme was not produced by any of the Iodic methods by 4 bacterial strains, while the rest of the species varied in the susceptibility of production to the enzyme. The Iodine tube is one of the best methods to detect the production of these enzymes. The susceptibility of bacterial species to the production of large-spectrum beta-lactase enzymes was also tested using the National Committee for Clinical Laboratory Standards (NCCL) and the double-disc method. Staphylococcus lentus, Staphylococcus capilis, Staphylococcus chromanogenes The first NCCL has shown a single isolation of Staphylococcus capilis ability to produce it.   http://dx.doi.org/10.25130/tjps.24.2019.125

scholarly journals Rapid differentiation of Staphylococcus aureus subspecies based on MALDI-TOF MS profiles

2018 ◽  
Vol 30 (6) ◽  
pp. 813-820 ◽  
Author(s):  
Marta Pérez-Sancho ◽  
Ana I. Vela ◽  
Pilar Horcajo ◽  
María Ugarte-Ruiz ◽  
Lucas Domínguez ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Rapid Identification ◽  
Automatic Identification ◽  
Identification System ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Different Origins ◽  
Tof Ms

Staphylococcus aureus encompasses 2 subspecies ( aureus and anaerobius) with significant differences in their epidemiology and pathogenicity. We evaluated the suitability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the rapid identification of both subspecies using a panel of 52 S. aureus isolates (30 subsp. anaerobius and 22 subsp. aureus) recovered from different origins, countries, and years. The on-board library identification system correctly identified 42 of 52 (81%) S. aureus isolates at the species level with score values >2.0. Limited performance was observed for differentiation of S. aureus subspecies (particularly subsp. anaerobius). Visual inspection of MALDI-TOF MS profiles identified 5 subspecies-specific mass peaks ( m/ z 3430 and 6861 in S. aureus subsp. anaerobius, and m/ z 4046, 6890, and 8093 in S. aureus subsp. aureus) with 100% sensitivity and specificity values, which is potentially useful for differentiating these subspecies. The suitability of 3 models, Genetic Analysis (GA), Quick Classifier (QC), and Supervised Neural Network, for automatic identification of both subspecies was evaluated using the Recognition Capability (RC) and Cross Validation (CV) values provided by the on-board ClinProTools software. The GA and QC models reached RC and CV values of 100%. Both models were externally validated using a panel of 26 S. aureus isolates of both subspecies, with both models correctly classifying all isolates of both subspecies. MALDI-TOF MS coupled with ClinProTools software represents a rapid and simple approach for S. aureus subspecies discrimination.

Evaluation of a Selective Broth for Detection of Staphylococcus aureus Using Impedance Microbiology

2001 ◽  
Vol 64 (1) ◽  
pp. 44-50 ◽  
Author(s):  
KIRSTEN E. GLASSMOYER ◽  
SCOTT M. RUSSELL
Keyword(s):  
Staphylococcus Aureus ◽  
Nalidixic Acid ◽  
Bacterial Species ◽  
Nutrient Broth ◽  
Microbiological Method ◽  
E Coli ◽  
Poultry Products ◽  
Chicken Carcass ◽  
Low Levels ◽  
Staphylococcus Spp

Experiments were conducted to evaluate a selective nutrient broth containing acriflavine and nalidixic acid for detection of Staphylococcus aureus using an impedance microbiological method. Nine species of bacteria, other than S. aureus, were evaluated using the selective broth to determine if these species could be inhibited. A total of 10 ppm of nalidixic acid inhibited the gram-negative species tested, with the exception of Pseudomonas aeruginosa. Similarly, 10 ppm of acriflavine suppressed the Staphylococcus spp. examined; however, S. aureus retained the ability to proliferate. Nutrient broth solution containing 10 ppm of nalidixic acid and 10 ppm of acriflavine (S. aureus impedance broth [SIB]) inhibited multiplication of most of the bacterial species tested and allowed S. aureus to be detected in an average of 16.4 h. Fresh chicken carcass rinses and cooked chicken rinses were inoculated with Escherichia coli and S. aureus and assayed using SIB in conjunction with impedance. Results demonstrated that S. aureus could be detected in less than 11.5 h, although the presence of E. coli decreased detection times. Additionally, impedance assays were conducted using five different poultry products to evaluate the sensitivity of the broth for detecting S. aureus. S. aureus could be detected on poultry products when present at low levels (101 CFU/ml) in less than 24 h. These studies demonstrated that SIB may be used in conjunction with impedance for rapid detection of S. aureus. However, without further modification, this method should not be used for enumeration of S. aureus from samples containing mixed microflora.

scholarly journals Molecular Identification of Staphylococcus spp Isolated from Clinical samples

2018 ◽  
Vol 12 (1) ◽  
pp. 14-24
Author(s):  
Taghreed Khudhur Mohammad
Keyword(s):  
Staphylococcus Aureus ◽  
Staphylococcus Epidermidis ◽  
Cultural Analysis ◽  
Clinical Samples ◽  
Microbiological Analysis ◽  
Sequencing Analysis ◽  
Staphylococcus Haemolyticus ◽  
Golden Yellow ◽  
Staphylococcus Spp ◽  
16Srrna Gene

The analysis of 16S rRNA gene sequences has been the technique generally used to study and confirm the identification and taxonomy of staphylococci. However, bacterial species cannot always be distinguished from each other using cultural methods. Thus,   clinical samples were collected from 190 cases only 31 positive for staphylococcal infections with Urinary Tract Infection, Wounds, Burns, Otitis media, diarrhea infections, were applied for microbiological analysis which include: cultures on Manitol salt agar and  HiCrome UTI Agar medium all the isolates gave positive golden yellow and identify as Staphylococcus spp. DNA was extracted from Staphylococcus spp and the 16srRNA gene were amplified by using specific primer, then sequencing of nucleic acid of genes was performed by machine is AB13730XL, Applied Biosystem, Macro gen company, the DNA sequencing results of flank sense of 16srRNA gene from 31 strains of Staphylococcus  was confirm the identification into species level: Staphylococcus haemolyticus, Staphylococcus aureus, Staphylococcus epidermidis And   Staphylococcus sciuri. Analysis of the sequences appeared that there two substitution(Transversion, Transition) in the Staphylococcus aureus strains with  Sequence ID LC090540.1 location at Range of nucleotide from 4 to 636, 100% compatibility with NCBI while no substitution appeared in the  Staphylococcus haemolyticus strains which have the sequence ID LN998078.1, 99% compatibility with NCBI also the sequence ID KR812401.1 which related to the strain Staphylococcus sciuri not appeared any substitution after sequencing analysis. Types of substitution detected in partial 16srRNA gene in Staphylococcus epidermidis strains 13  Transversion  and 5 transition substitution location at range of nucleotide from 6 to 1026 have the  Sequence ID KF575160.1 compared with data obtained from Gene Bank, these finding lead to conclusion, our assay allows rapid and confirm the detection to avoid possibility of misidentification of Staphylococcus species based on cultural analysis, the study aimed to  propose the partial sequencing of the gene as an alternative molecular tool for the analysis of Staphylococcus species and for decreasing the possibility of misidentification. New submission of local Iraqi Staphylococcus clinical isolated during the current study show successfully record of four isolate Staphylococcus sciuri, Staphylococcus epidermidis, Staphylococcus aureus and Staphylococcus haemolyticus with GenBank accession number: KY938530.1 ,KY938529.1, ,KY938528.1, and  GenBank: KY938527.1respectivelly.

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