Updating Molecular Diagnostics for Detecting Methicillin-Susceptible and Methicillin-Resistant Staphylococcus aureus Isolates in Blood Culture Bottles.

Molecular diagnostic tests can be used to provide rapid identification of staphylococcal species in blood culture bottles to help improve antimicrobial stewardship. However, alterations in the target nucleic acid sequences of the microorganisms or their antimicrobial resistance genes can lead to false-negative results. We determined the whole-genome sequences of 4 blood culture isolates of and 2 control organisms to understand the genetic basis of genotype-phenotype discrepancies when using the Xpert MRSA/SA BC test ( diagnostic medical device [IVD]). Three methicillin-resistant (MRSA) isolates each had a different insertion of a genetic element in the staphylococcal cassette chromosome (SCC)- junction region that led to a misclassification as methicillin-susceptible (MSSA). One strain contained a deletion in , which produced a false -negative result. A control strain of that harbored an SCC element but no (an empty cassette) was correctly called MSSA by the Xpert test. The second control contained an SCC insertion. The updated Xpert MRSA/SA BC test successfully detected both and SCC variants of MRSA and correctly identified empty-cassette strains of as MSSA. Among a sample of 252 MSSA isolates from the United States and Europe, 3.9% contained empty SCC cassettes, 1.6% carried SCC, <1% had deletions, and <1% contained SCC variants other than those with SCC These data suggest that genetic variations that may interfere with Xpert MRSA/SA BC test results remain rare. Results for all the isolates were correct when tested with the updated assay.