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Updated: Mar 29, 2026
Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria
Published on: May 8, 2013
Comparison between a chimeric lysin ClyH and other enzymes for extracting DNA to detect methicillin resistant
Yuanyuan Hu1,2, Hang Yang1, Jing Wang1
1Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Abstract:
Extracting DNA from Staphylococcus aureus cells is important for detecting MRSA by PCR. However, S. aureus cells are known to be difficult to disrupt due to their compact cell walls. Here, we systematically studied the efficiency of a highly active lysin ClyH for extracting DNA of S. aureus in comparison with commonly used enzymes, such as lysostaphin and achromopeptidase (ACP), and its compatibility in quantitative PCR (qPCR) detection of MRSA. qPCR analysis of S. aureus specific gene femB showed that ClyH was much faster than lysostaphin, ACP and lysozyme for releasing DNA. Five minutes disruption with ClyH at room temperature was enough to release all the DNA from S. aureus. Analysis of the spiked nasal swabs by a dual qPCR assay of the β-lactam resistance mecA gene and the staphylococcal cassette chromosome (SCCmec)-open reading frame X (orfX) junction (SCCmec-orfX) after ClyH lysis showed 100% sensitivity and specificity to the commercial BD GeneOhm™ MRSA test with ACP lysis, but the lysis time was reduced from 20 min by ACP to 5 min by ClyH. Our research shows that ClyH could be a better option than the currently used enzymes for DNA extraction from S. aureus, which can provide simpler and faster PCR detection of MRSA.
Insights
A novel enzyme, ClyH, efficiently extracts DNA from Staphylococcus aureus cells in just five minutes, significantly speeding up Methicillin-resistant Staphylococcus aureus (MRSA) detection.
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- Staphylococcus aureus possesses robust cell walls, complicating DNA extraction for diagnostics.
- Efficient DNA extraction is crucial for accurate and rapid detection of Methicillin-resistant Staphylococcus aureus (MRSA).
Purpose of the Study:
- To evaluate the efficiency of the lysin ClyH for Staphylococcus aureus DNA extraction.
- To compare ClyH with conventional enzymes like lysostaphin and achromopeptidase (ACP).
- To assess ClyH's compatibility with quantitative PCR (qPCR) for MRSA detection.
Main Methods:
- Systematic study of ClyH's DNA extraction efficiency on Staphylococcus aureus.
- Comparative analysis with lysostaphin, achromopeptidase (ACP), and lysozyme.
- Dual qPCR assay for mecA and SCCmec-orfX detection using ClyH-lysed samples.
Main Results:
- ClyH demonstrated significantly faster DNA release from Staphylococcus aureus compared to lysostaphin, ACP, and lysozyme.
- Five minutes of ClyH disruption at room temperature was sufficient for complete DNA release.
- ClyH lysis in a dual qPCR assay achieved 100% sensitivity and specificity for MRSA detection, reducing lysis time from 20 to 5 minutes.
Conclusions:
- ClyH is a highly efficient enzyme for Staphylococcus aureus DNA extraction.
- ClyH offers a faster and simpler alternative to current enzymes for MRSA PCR detection.
- The use of ClyH can significantly improve the speed and efficiency of MRSA diagnostic assays.
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