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Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization
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| Zeitschriftentitel: | Journal of Clinical Microbiology |
|---|---|
| Personen und Körperschaften: | , , , , , , , , , |
| In: | Journal of Clinical Microbiology, 51, 2013, 9, S. 2869-2874 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society for Microbiology
|
| Schlagwörter: |
| author_facet |
Yam, W. C. Siu, Gilman K. H. Ho, P. L. Ng, T. K. Que, T. L. Yip, K. T. Fok, Cathie P. K. Chen, Jonathan H. K. Cheng, Vincent C. C. Yuen, K. Y. Yam, W. C. Siu, Gilman K. H. Ho, P. L. Ng, T. K. Que, T. L. Yip, K. T. Fok, Cathie P. K. Chen, Jonathan H. K. Cheng, Vincent C. C. Yuen, K. Y. |
|---|---|
| author |
Yam, W. C. Siu, Gilman K. H. Ho, P. L. Ng, T. K. Que, T. L. Yip, K. T. Fok, Cathie P. K. Chen, Jonathan H. K. Cheng, Vincent C. C. Yuen, K. Y. |
| spellingShingle |
Yam, W. C. Siu, Gilman K. H. Ho, P. L. Ng, T. K. Que, T. L. Yip, K. T. Fok, Cathie P. K. Chen, Jonathan H. K. Cheng, Vincent C. C. Yuen, K. Y. Journal of Clinical Microbiology Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization Microbiology (medical) |
| author_sort |
yam, w. c. |
| spelling |
Yam, W. C. Siu, Gilman K. H. Ho, P. L. Ng, T. K. Que, T. L. Yip, K. T. Fok, Cathie P. K. Chen, Jonathan H. K. Cheng, Vincent C. C. Yuen, K. Y. 0095-1137 1098-660X American Society for Microbiology Microbiology (medical) http://dx.doi.org/10.1128/jcm.00488-13 <jats:title>ABSTRACT</jats:title> <jats:p> Rapid detection of methicillin-resistant <jats:named-content content-type="genus-species">Staphylococcus aureus</jats:named-content> (MRSA) nasal colonization is crucial for the prevention and control of MRSA infections in health care settings. The LightCycler MRSA Advanced Test (Roche Diagnostics) is a commercially available real-time PCR assay for direct detection of MRSA nasal colonization by targeting of the staphylococcal cassette chromosome <jats:italic>mec</jats:italic> (SCC <jats:italic>mec</jats:italic> )- <jats:italic>orfX</jats:italic> junction. The diagnostic performance of the assay was compared with that of ChromID MRSA agar (bioMérieux) culture and an in-house duplex real-time PCR assay. Among 1,246 nasal swab specimens collected from 2 general hospitals in Hong Kong, 174 (14%) were considered true positive for MRSA. Chromogenic culture and the in-house real-time PCR assay identified 147 (84.5%) and 133 (76.4%) true-positive cases with specificities of 100% and 98.6%, respectively. Based on the target melting temperature ( <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> ) values (57.0 to 62.0°C) defined by the manufacturer, the LightCycler MRSA Advanced Test identified only 85 (48.9%) true-positive specimens. Interestingly, an additional 60 (34.5%) true-positive specimens were detected despite atypical <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C, providing overall sensitivity and specificity values of 83.3% and 99%, respectively. Among isolates with <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C, most were typed as clonal complex 45 (CC45). By sequence analysis of the SCC <jats:italic>mec-orfX</jats:italic> junction, characteristic single-nucleotide polymorphisms (SNPs) were identified only in isolates with <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C and not in those with typical <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values. It is conceivable that those SNPs were located inside the target region of the proprietary hybridization probes, which resulted in a <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> shift in the melting curve analysis. Our study highlights the importance of a global evaluation of commercial kits so that the interpretation algorithm covers different lineages of MRSA clones prevalent in various geographical regions. </jats:p> Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization Journal of Clinical Microbiology |
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American Society for Microbiology, 2013 |
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2013 |
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American Society for Microbiology |
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Journal of Clinical Microbiology |
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49 |
| title |
Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_unstemmed |
Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_full |
Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_fullStr |
Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_full_unstemmed |
Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_short |
Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_sort |
evaluation of the lightcycler methicillin-resistant staphylococcus aureus (mrsa) advanced test for detection of mrsa nasal colonization |
| topic |
Microbiology (medical) |
| url |
http://dx.doi.org/10.1128/jcm.00488-13 |
| publishDate |
2013 |
| physical |
2869-2874 |
| description |
<jats:title>ABSTRACT</jats:title>
<jats:p>
Rapid detection of methicillin-resistant
<jats:named-content content-type="genus-species">Staphylococcus aureus</jats:named-content>
(MRSA) nasal colonization is crucial for the prevention and control of MRSA infections in health care settings. The LightCycler MRSA Advanced Test (Roche Diagnostics) is a commercially available real-time PCR assay for direct detection of MRSA nasal colonization by targeting of the staphylococcal cassette chromosome
<jats:italic>mec</jats:italic>
(SCC
<jats:italic>mec</jats:italic>
)-
<jats:italic>orfX</jats:italic>
junction. The diagnostic performance of the assay was compared with that of ChromID MRSA agar (bioMérieux) culture and an in-house duplex real-time PCR assay. Among 1,246 nasal swab specimens collected from 2 general hospitals in Hong Kong, 174 (14%) were considered true positive for MRSA. Chromogenic culture and the in-house real-time PCR assay identified 147 (84.5%) and 133 (76.4%) true-positive cases with specificities of 100% and 98.6%, respectively. Based on the target melting temperature (
<jats:italic>
T
<jats:sub>m</jats:sub>
</jats:italic>
) values (57.0 to 62.0°C) defined by the manufacturer, the LightCycler MRSA Advanced Test identified only 85 (48.9%) true-positive specimens. Interestingly, an additional 60 (34.5%) true-positive specimens were detected despite atypical
<jats:italic>
T
<jats:sub>m</jats:sub>
</jats:italic>
values of 55°C, providing overall sensitivity and specificity values of 83.3% and 99%, respectively. Among isolates with
<jats:italic>
T
<jats:sub>m</jats:sub>
</jats:italic>
values of 55°C, most were typed as clonal complex 45 (CC45). By sequence analysis of the SCC
<jats:italic>mec-orfX</jats:italic>
junction, characteristic single-nucleotide polymorphisms (SNPs) were identified only in isolates with
<jats:italic>
T
<jats:sub>m</jats:sub>
</jats:italic>
values of 55°C and not in those with typical
<jats:italic>
T
<jats:sub>m</jats:sub>
</jats:italic>
values. It is conceivable that those SNPs were located inside the target region of the proprietary hybridization probes, which resulted in a
<jats:italic>
T
<jats:sub>m</jats:sub>
</jats:italic>
shift in the melting curve analysis. Our study highlights the importance of a global evaluation of commercial kits so that the interpretation algorithm covers different lineages of MRSA clones prevalent in various geographical regions.
</jats:p> |
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| author | Yam, W. C., Siu, Gilman K. H., Ho, P. L., Ng, T. K., Que, T. L., Yip, K. T., Fok, Cathie P. K., Chen, Jonathan H. K., Cheng, Vincent C. C., Yuen, K. Y. |
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| description | <jats:title>ABSTRACT</jats:title> <jats:p> Rapid detection of methicillin-resistant <jats:named-content content-type="genus-species">Staphylococcus aureus</jats:named-content> (MRSA) nasal colonization is crucial for the prevention and control of MRSA infections in health care settings. The LightCycler MRSA Advanced Test (Roche Diagnostics) is a commercially available real-time PCR assay for direct detection of MRSA nasal colonization by targeting of the staphylococcal cassette chromosome <jats:italic>mec</jats:italic> (SCC <jats:italic>mec</jats:italic> )- <jats:italic>orfX</jats:italic> junction. The diagnostic performance of the assay was compared with that of ChromID MRSA agar (bioMérieux) culture and an in-house duplex real-time PCR assay. Among 1,246 nasal swab specimens collected from 2 general hospitals in Hong Kong, 174 (14%) were considered true positive for MRSA. Chromogenic culture and the in-house real-time PCR assay identified 147 (84.5%) and 133 (76.4%) true-positive cases with specificities of 100% and 98.6%, respectively. Based on the target melting temperature ( <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> ) values (57.0 to 62.0°C) defined by the manufacturer, the LightCycler MRSA Advanced Test identified only 85 (48.9%) true-positive specimens. Interestingly, an additional 60 (34.5%) true-positive specimens were detected despite atypical <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C, providing overall sensitivity and specificity values of 83.3% and 99%, respectively. Among isolates with <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C, most were typed as clonal complex 45 (CC45). By sequence analysis of the SCC <jats:italic>mec-orfX</jats:italic> junction, characteristic single-nucleotide polymorphisms (SNPs) were identified only in isolates with <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C and not in those with typical <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values. It is conceivable that those SNPs were located inside the target region of the proprietary hybridization probes, which resulted in a <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> shift in the melting curve analysis. Our study highlights the importance of a global evaluation of commercial kits so that the interpretation algorithm covers different lineages of MRSA clones prevalent in various geographical regions. </jats:p> |
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| spelling | Yam, W. C. Siu, Gilman K. H. Ho, P. L. Ng, T. K. Que, T. L. Yip, K. T. Fok, Cathie P. K. Chen, Jonathan H. K. Cheng, Vincent C. C. Yuen, K. Y. 0095-1137 1098-660X American Society for Microbiology Microbiology (medical) http://dx.doi.org/10.1128/jcm.00488-13 <jats:title>ABSTRACT</jats:title> <jats:p> Rapid detection of methicillin-resistant <jats:named-content content-type="genus-species">Staphylococcus aureus</jats:named-content> (MRSA) nasal colonization is crucial for the prevention and control of MRSA infections in health care settings. The LightCycler MRSA Advanced Test (Roche Diagnostics) is a commercially available real-time PCR assay for direct detection of MRSA nasal colonization by targeting of the staphylococcal cassette chromosome <jats:italic>mec</jats:italic> (SCC <jats:italic>mec</jats:italic> )- <jats:italic>orfX</jats:italic> junction. The diagnostic performance of the assay was compared with that of ChromID MRSA agar (bioMérieux) culture and an in-house duplex real-time PCR assay. Among 1,246 nasal swab specimens collected from 2 general hospitals in Hong Kong, 174 (14%) were considered true positive for MRSA. Chromogenic culture and the in-house real-time PCR assay identified 147 (84.5%) and 133 (76.4%) true-positive cases with specificities of 100% and 98.6%, respectively. Based on the target melting temperature ( <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> ) values (57.0 to 62.0°C) defined by the manufacturer, the LightCycler MRSA Advanced Test identified only 85 (48.9%) true-positive specimens. Interestingly, an additional 60 (34.5%) true-positive specimens were detected despite atypical <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C, providing overall sensitivity and specificity values of 83.3% and 99%, respectively. Among isolates with <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C, most were typed as clonal complex 45 (CC45). By sequence analysis of the SCC <jats:italic>mec-orfX</jats:italic> junction, characteristic single-nucleotide polymorphisms (SNPs) were identified only in isolates with <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values of 55°C and not in those with typical <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> values. It is conceivable that those SNPs were located inside the target region of the proprietary hybridization probes, which resulted in a <jats:italic> T <jats:sub>m</jats:sub> </jats:italic> shift in the melting curve analysis. Our study highlights the importance of a global evaluation of commercial kits so that the interpretation algorithm covers different lineages of MRSA clones prevalent in various geographical regions. </jats:p> Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization Journal of Clinical Microbiology |
| spellingShingle | Yam, W. C., Siu, Gilman K. H., Ho, P. L., Ng, T. K., Que, T. L., Yip, K. T., Fok, Cathie P. K., Chen, Jonathan H. K., Cheng, Vincent C. C., Yuen, K. Y., Journal of Clinical Microbiology, Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization, Microbiology (medical) |
| title | Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_full | Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_fullStr | Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_full_unstemmed | Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_short | Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| title_sort | evaluation of the lightcycler methicillin-resistant staphylococcus aureus (mrsa) advanced test for detection of mrsa nasal colonization |
| title_unstemmed | Evaluation of the LightCycler Methicillin-Resistant Staphylococcus aureus (MRSA) Advanced Test for Detection of MRSA Nasal Colonization |
| topic | Microbiology (medical) |
| url | http://dx.doi.org/10.1128/jcm.00488-13 |