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Detection of methicillin-resistant Staphylococcus aureus in donor eye preservation media by polymerase chain reaction
1Department of Ophthalmology, University of Tokyo School of Medicine, Japan.
Abstract:
To examine contamination of donor eye preservation media with methicillin-resistant Staphylococcus aureus (MRSA), we studied the media to detect specific genes for MRSA by use of the polymerase chain reaction (PCR) method. The materials were 36 samples of donor eye preservation media (EP-II) in which donor eyes had been stored for keratoplasty. Polymerase chain reaction procedures were carried out to simultaneously detect the spa gene, which codes protein A of S. aureus, and the mecA gene, which codes penicillin-binding protein-2' contributing resistance to methicillin. Along with PCR analyses, the preservation media was examined by conventional culture methods to determine bacteriologic contamination. The PCR analyses of the 36 samples revealed that both the spa and the mecA genes were positive in five samples, only the spa gene was positive in two, and only the mecA gene was positive in two. The conventional culture of the media showed positive for MRSA in 5 samples, methicillin-susceptible S. aureus (MSSA) in 2, methicillin-susceptible coagulase-negative staphylococci (MS-CNS) in 4, and methicillin-resistant coagulase-negative staphylococci (MR-CNS) in 2 of the 36 samples. The results of PCR coincided well with those of conventional bacteriologic culture. Polymerase chain reaction analysis for spa and mecA genes is useful in detecting contamination of donor eye preservation media by MRSA, MSSA, MR-CNS, or MS-CNS in a shorter time than by conventional culture.
Insights
This study investigated donor eye preservation media for bacterial contamination using PCR. Polymerase chain reaction (PCR) effectively detected methicillin-resistant Staphylococcus aureus (MRSA) and other staphylococci, offering a faster alternative to traditional culture methods.
Area of Science:
- Ophthalmology
- Microbiology
- Molecular Biology
Background:
- Donor eye preservation media is crucial for keratoplasty success.
- Bacterial contamination of this media poses a risk to recipients.
- Rapid detection methods are needed to ensure donor tissue safety.
Purpose of the Study:
- To evaluate the utility of polymerase chain reaction (PCR) for detecting methicillin-resistant Staphylococcus aureus (MRSA) in donor eye preservation media.
- To compare PCR-based detection with conventional bacteriologic culture methods.
Main Methods:
- 36 samples of donor eye preservation media (EP-II) were analyzed.
- PCR was used to detect the spa and mecA genes specific to S. aureus and methicillin resistance.
- Conventional culture methods were employed for bacteriologic examination.
Main Results:
- PCR detected both spa and mecA genes in 5 samples, indicating MRSA contamination.
- PCR identified other staphylococcal contaminations (MSSA, MR-CNS, MS-CNS).
- PCR results correlated well with conventional culture findings, but offered faster results.
Conclusions:
- PCR is a valuable tool for rapid and accurate detection of MRSA and other staphylococcal contaminants in donor eye preservation media.
- This molecular method can significantly reduce the time required for screening, enhancing donor eye safety.