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Jorge Maestre, Martha Diaz, Edith Perez, Harry W Flynn, Eduardo C Alfonso, Darlene Miller, ; Comparison of the Automated Repetitive Sequence-Based PCR Microbial Typing System (DiversiLab) vs Multiplex PCR for Molecular Characterization of Ocular MRSA.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4513.
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MRSA among ocular isolates continue to increase in frequency and morbidity. Rapid Molecular characterization provides information on origin, distribution, therapeutic choices and transmission. We evaluated the DiversiLab rep-PCR Microbial typing system as a rapid method for documenting the molecular profiles and clonality of ocular MRSA strains to provide a better understanding of ocular MRSA epidemiology and transmission.
We compared the use of multiplex PCR with the commercial DiversLab (DL) to identify and discriminate between SCCmec types among 56 MRSA isolates from 6 ocular sources (cornea, N=23, conj or lids, N=14, lacrimal sac, N=10, socket, N=5, vitreous-N=4). Similarity index (% agreement) and feasibility of using the DL system for spa typing and clonality (origin) of ocular MRSA isolates were documented against 2 well characterized, standard ATCC MRSA research panels organized by SCCmec and or PFGE types
: Concordance of SCCmec types of the two systems was 64.3%. Distribution of SCCmec types via multiplex PCR was: Type II (MRSA-HA), 64.3%,Type IV (MRSA-CA), 23.2% Type V, 3.6% Nontypable, 8.9%, DL profiles included: Type I (MRSA-HA), 16.1%, Type II, 35.7%, Type III (MRSA-CA), 8.9%, Type IV, 35.7% and Nontypable, 3.6%. The greatest discord was for SCCmec type II. Type II isolates were more frequently recovered from lacrimal sac 90% and cornea- 65% cultures. Type IV was most frequently recovered from conj/lids/socket 57.9%. DL divided ocular isolates into 5 clonal complexes, CC8 (41.1%)-which houses epidemic MRSA (EMRSA) community strains, CC5 (30.3%)-housing USA100-the most common healthcare associated-MRSA strain in the US, CC30 (19.6%)-USA200 and EMRSA 16-second most common MRSA-HA strain, CC22 (5.3%)-EMRSA-15 and-CC80 (3.6%)-an emerging ocular MRSA strain. The 5 sequence types (ST), ST5, ST254, ST228, ST30 and ST247 ranged from 8.9% (ST247) to 25% (ST5). Agreement with the standard ATCC panels was 87.5% (7/8).
Ocular MRSA have diverse clonal origins and may be site specific. The DiversiLab provides a rapid, method for screening ocular MRSA, determining evolutionary origins and documenting the emergence of unique MRSA strains.
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