scholarly journals Molecular Typing of Methicillin-Resistant Staphylococcus aureus by Polymerase Chain Reaction: Distribution of the Typed Strains in Hospitals.

1997 ◽  
Vol 36 (10) ◽  
pp. 694-699 ◽  
Author(s):  
Shuji TOHDA ◽  
Miki MARUYAMA ◽  
Nobuo NARA
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Molecular Typing ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Chain Reaction ◽  
Methicillin Resistant ◽  
Polymerase Chain

scholarly journals Molecular Typing of Methicillin-sensitive and Methicillin-resistant Staphylococcus aureus: What Epidemiologists need to know from Plasmid Analysis to Whole Genome Sequencing.

2016 ◽  
Vol 6 (1) ◽  
pp. 31-43
Author(s):  
Ibrahim A. Al-Zahrani
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Molecular Typing ◽  
Gel Electrophoresis ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Pulsed Field Gel Electrophoresis ◽  
Whole Genome ◽  
Chain Reaction ◽  
Methicillin Resistant ◽  
Polymerase Chain

Methicillin-resistant Staphylococcus aureus is an important nosocomial pathogen. The early identification of an outbreak, making use of a rapid, precise and simple methicillin-resistant Staphylococcus aureus typing technique, can lead to prompt and effective precautions that avoid further spread of the infection. Pulsed-field gel electrophoresis is considered the gold standard for methicillin-resistant Staphylococcus aureus typing and has been recently supported by multilocus sequence typing. SmaI-multiplex polymerase chain reaction typing is a novel polymerase chain reaction -based technique that combines the principles of pulsed-field gel electrophoresis, with simplicity of polymerase chain reaction and can be used in many routine clinical laboratories. Whole genome sequencing typing is a second generation sequencing technology and may ultimately replace the traditional molecular typing methods. This review aims to survey existing molecular typing techniques for Staphylococcus aureus and to discuss information for each method in order to aid researchers and clinical professionals in the selection and implementation of an optimal technique.

Polymerase Chain Reaction for the Rapid Detection of Cerebrospinal Fluid Shunt or Ventriculostomy Infections

Neurosurgery ◽  
2005 ◽  
Vol 57 (6) ◽  
pp. 1237-1243 ◽  
Author(s):  
Jason T. Banks ◽  
Suman Bharara ◽  
R Shane Tubbs ◽  
Charles L. Wolff ◽  
G Yancey Gillespie ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Cerebrospinal Fluid ◽  
16S Rrna ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Propionibacterium Acnes ◽  
Chain Reaction ◽  
Gram Negative ◽  
Methicillin Resistant ◽  
Polymerase Chain

AbstractOBJECTIVE:Infection after cerebrospinal fluid (CSF) shunts or ventriculostomies is a common complication associated with significant morbidity and mortality. Polymerase chain reaction (PCR) is a powerful molecular technique that allows rapid and precise amplification of bacterial deoxyribonucleic acid (DNA) and has proven a powerful tool in the detection of a wide variety of clinically important infectious diseases. We analyzed specimens of CSF derived from ventriculoperitoneal shunts or external ventricular drains by using both conventional cultures and PCR and report herein our preliminary results.METHODS:We selected 86 CSF samples from adult patients who underwent either shunt tap or routine surveillance cultures of their ventriculostomy. These specimens were chosen from a larger group of 300 specimens that were routinely collected (many serially) in our clinical practice. They were chosen because clinical suspicion of infection was increased because of either patient signs and symptoms (fever, stiff neck, lethargy, worsening neurological examination) or preliminary laboratory analysis of CSF data (increased white blood cell count, increased protein level, decreased glucose). We considered this subgroup optimal to efficiently initiate our investigation of the correlation of PCR and culture results. CSF was increased by using standard culture techniques and by using PCR. Samples of CSF that were to undergo PCR had DNA extracted, purified, and amplified for 16S rRNA using primers 16S-Forward and 16S-Reverse of conserved sequence regions of all bacteria. DNA was PCR-amplified for 30 cycles. One microliter of the first PCR product was subjected to nested PCR using primers specific for gram-positive and gram-negative bacteria. Samples were also subjected to PCR amplification for specific detection of Propionibacterium acnes, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus using specific primers for 16S rRNA Propionibacterium, nuclease gene of Staphylococcus, and Mec gene of methicillin-resistant Staphylococcus aureus.RESULTS:For 18 of 86 specimens (21%), both the culture and PCR were positive. For 30 of 86 specimens (35%), both the PCR and culture results were negative. For 42 of 86 specimens (49%), cultures were negative and PCR was positive. There were no positive culture results with negative PCR results. Most negative culture/positive PCR cases occurred after prolonged intravenous antibiotics. Of the 56 PCR-positive specimens, 30 were positive for Propionibacterium acnes, whereas 40 were positive for Staphylococcus aureus. Of the Staphylococcus aureus-positive specimens, two were positive for methicillin resistant-Staphylococcus aureus. Among the 56 PCR-positive specimens, 30 were positive for both Propionibacterium acnes and Staphylococcus aureus; gram-negative organisms were not detected by any method in these specimens.CONCLUSION:These preliminary data suggest that PCR is a highly sensitive, rapid, and potentially promising modality for the detection and treatment of CSF shunt ventriculostomy infection.

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